New Modes of GPCR Signalling

INVITED LIST OF INVITED SPEAKER’S SESSION SPEAKER’S SPEAKER’S ABSTRACT
ABSTRACT
ABSTRACT
New Modes of GPCR Signalling ……………………………………………………… 20
Joel Bockaert
Regulation of Cellular Metabolism by Protein Lysine
Acetylation…………………………23
Kun-Liang Guan
Sensing mGlu Receptor Activation…………………………………………………….......26
Jean-Philippe Pin
mGlu Receptors : Complex Allosteric Machines to Tune Up Synaptic
Transmission………...29
Philippe Rondard
Manipulation of Rac Activity and Cell Motility in vivo Using
Light……………………...32
Denise J. Montell
Activation Mechanism of GABAB Receptor………………………………………………35
Jianfeng Liu
From Bench to Bedside: BACE1, the Beta-Secretase Enzyme, in Basic Science and
Clinical
Investigation……………………………………………………………………………..38
Yong Shen
TRPA1: a Sentinel for Noxious Chemicals and
Temperatures……………………………...41
Craig Montell
Novel Genes and Pathways that Regulate Alzheimer’s Disease
Pathogenesis……………….44
Huaxi Xu
Structural Feature and Function of hβ-subunits of BK Channel…………………………43
Jiuping Ding
The Synaptic Mechanisms of Glycogen Synathese Kinase-3 in Regulating Leaning and
Memory………………………………………………………………………………….50
Jianzhi Wang
Sensory Transduction in C. elegans: What Can't a Worm
Sense?................................................53
Shawn X. Xu
Spatial Control of Synaptic Transmission at a Cholinergic Synapse in C.
elegans………..56
Jean-Louis Bessereau
The Function of Phosphatidylinositol-Linked Novel D1 Dopamine Receptor in Central
Nervous System…………………………………………………………………………..59
Jian-Guo Chen

INVITED SESSION ABSTRACT
Mechanisms of Wnt Signaling and Regulation…………………………………………...62
Xi He
Acid Sensing Ion Channels: A Novel Therapeutic Target for Anxiety and
Depression?............65
John Wemmie
Sequential Activation of Orai1 Subunits by Different Numbers of STIM1 Leads to
Multistate Opening of the CRAC Channel………………………………………………..68
Tao Xu

LIST OF SOLICITED ABSTRACT
Chemosensory and Behavior of C.elegans is Regulated by An E.Coli Noncoding RNA
..................................72
Implication of the env Gene of the Human Endogenous Retrovirus W Family in the
Expression of BDNF and DRD3
..................................73
Microvavle Immobilization of Intact C. Elegans for in vivo Calcium Imaging of
Neuronal Activities in Response to External Stimuli
..................................74
Inhibitory Effect of Ethanol on Gustatory Plasticity in C. Elegans Mediated by the
Serotonin Pathway
..................................75
Isoform-Specific Prolongation of KCNQ (Kv7) Potassium Channel Opening Mediated
by a New Drug-Channel Binding Site
..................................76
Novel USH2A Compound Heterozygous Mutations Cause RP/USH2 in a Chinese
Family
..................................77
UNC-31/CAPS Docks and Primes Dense Core Vesicles in C. elegans Neurons
..................................78
Cordycepin Protects Against Brain ischemical Reperfusion Injury in Mice by Inhibition
of Matrix Metalloproteinases-3 Expression.
..................................79
Calcium Signaling in Astrocytes Induced by Photostimulation with Femtosecond Laser
..................................80
ZD7288 Inhibits the Induction Of LTP in an NMDA Independent Manner at
Hippocampal Schaffer Collateral-CA1 Synapses
..................................81
Isolation, Characterization and Anti-Cancer Activity of SK84, a Novel Glycine-Rich
Antimicrobial Peptide from Drosophila Virilis
..................................82
Foamy Virus as Potential Vectors for Gene Therapy in Nervous Disorders and Signal
Pathway in Neural Cell Transduction
..................................83
Role of Heat on the Expression and Function of KCNQ2
..................................84
Possible Involvement of TRPV1 in neonatal Febrile Seizure Mice
..................................85
The Kv1.3 Channel Blocker Scorpion Polypeptide has an Therapeutic effect on EAE
Rats with a Molecule Mechanism
..................................86
IGF-1 Promote BPH-1 Cell Proliferation Primarily via Activating mTOR-dependent
Translational Increases in Cyclin D Proteins

..................................88
GABAB Receptor-Mediated Rap1 Activation and GB1/Rap1GTP Interaction in Neuronal
System
..................................89
Therapeutic Effect of Flupirtine on RepetitiveFebrile Seizures Rats
..................................90
Rapid Detection of Deletion of the PMP22 Gene from a Kindred in China with
Hereditary Neuropathy with Liability to Pressure Palsy by Real-time Quantitative PCR
..................................91
Therapeutic Effect of Flupirtine on Repetitive Febrile Seizures Rat
..................................92
TEF3(Transcription Enhancer Factor 3)Induced Angiogenesis through VEGF Pathway
..................................93
Does the Metabotropic Glutamate Receptor 7 (Mglur7) Play a Role in Schizophrenia?
..................................94
Effects of Yishendaluo Decoction on Axonal Degeneration, Inflammatory Reaction, and
Neurological Function in a Mouse Model of Experimental Autoimmune
Encephalomyelitis
..................................95
Imaging Neuronal Population in Vitro with Acousto-Optic Deflector Based Random
Access Two-Photon Microscopy
..................................96
Inhibition of Protein Phosphatase 2A Activity Plays a Key Role for Normal Human
Cells to Acquire TRAIL-Sensitive Phenotype during Tumorigenesis
..................................97
A Novel Mutation of PAX3 on a Large Chinese Family with Waardenburg Syndrome
..................................98
Identification a Novel MYO6 Mutation Associated with Autosomal Dominant
Non-Syndromic Hearing Impairment in a Large Chinese Family
..................................99
MAPK Scaffolding by BIT1 in the Golgi Complex Modulates Stress Resistance
................................100
Identification of a Novel Genetic Locus on Chromosome 8p21.1-q11.23 for Idiopathic
Basal Ganglia Calcification
................................101
Phosphoinositide-3-Kinase Pathway Activation in PTEN Deficient Prostate Cancer
Cells is Independent of Receptor Tyrosine Kinases and Mediated by the p110 or p110
Catalytic Subunits
................................102
Insulin-Like Signaling Pathway Functions in Integrative Response to an Olfactory and
a Gustatory Stimuli in Caenorhabditis elegans
................................103
Akt Overexpression Opposes the Beta Amyloid Toxic Injuries to HEK293/tau Cells
................................104
NMDA Receptors Subunit NR2B Messenger Rnas Expression with Cognitive
Dysfunction after Bilateral Common Carotid Artery Occlusion in the Adult Rats

................................105
Effects of ZD7288 on Long-Term Potentiation at Perforant Pathway(PP) Fibers - CA3
Region Synapse Pathway in Rat Hippocampus in Vivo
................................106
Effects of Cscl on Synaptic Transmission at Perforant Pathway (PP) Fibers - CA3
Region of Hippocampus in Rat
................................107
Embryonic Heart Growth is Regulated by Functional Antagonism Between GRIDLOCK
and GATA5 in Zebrafish
................................108
Quantitative Phosphoproteome Analysis Reveals Signaling Events Involved in
Aroma-Stimulated Mouse Olfactory Bulbs
................................109
Quantitative Phosphoproteomics Analysis of GABAB Receptor Signaling
................................110
Autotaxin Signaling via Lysophosphatidic Acid Receptors Contributes to LPC-Induced
Vascular Smooth Muscle Cell Proliferation and Migration
................................111
Morphine Regulated Thioredoxin-1 Expression in Neuroblastoma SH-SY5Y Cells
................................112
Cyclin D1 Promotes Anchorage-independent Cell Survival by Inhibiting
FOXO-Mediated Anoikis
................................113
Investigate the Role of Rab27 in the Dcvs Docking and Priming Process in C. elegans
................................114
Biophotons as Neural Communication Signals
................................115
Effects of Cyclic Nucleotide Gated Channel Knockout on the Olfactory Bulb Analyzed
by Quantitative Phospho-Proteomics
................................116

INVITED
SPEAKER’S
ABSTRACTS

INVITED SPEAKER’S INFO
Prof. Joël Bockaert
Professor , Director of Institut for Functional
Genomics(IGF)Montpellier
UMR5203 – INSERM U661 – UMI - UMII
141 Rue de la Cardonille 34094
Montpellier Cedex 5 FRANCE
Tel : (+33) 4 67 14 29 30
Fax : (+33) 4 67 14 29 10
E-mail: joel.bockaert@igf.cnrs.fr
Education
1961 Entrance examination for the Ecole Normale d'Instituteurs in Mérignac
1963 Baccalauréat of Secondary Education : Experimental Science
1964 Physical, Chemical and Natural Sciences (SPCN) at Université de Bordeaux
1966 Licence ès Sciences
1967 Diplôme d'Etudes Supérieures in Natural Sciences
1968 Agrégation in Natural Sciences
1973 Doctorate in Natural Sciences - Honorable distinction
Selected Publications over 497
1. Sladeczek F., Pin J.P., Récasens M., Bockaert J.S.Weiss (1985) Glutamate
stiumates inositol phosphate formation in striatal neurones. Nature 317: 717-719.
611 citations
2. Harris-Warrick R., Hammond C., Paupardin-Tritsch D., Homburger V., Rouot B. et
Bockaert J. (1988) The a subunit of GTP binding protein homologous to
mammalina Goa mediates a dopamine-induced decrease of calcium current in snail
neurons. Neuron 1: 27-32 120 citations
3. Dumuis A., Sebben M., Haynes L., Pin .P. et Bockaert J. (1988) NMDA receptors
activate the arachidonic acid cascade system in striatal neurons. Nature 336: 68-70.
479 citations
4. Dumuis A, Pin JP, Oomagari K, Sebben M, Bockaert J (1990) Arachidonic acid
released from striatal neurons by joint stimulation of ionotropic and metabotropic
quisqualate receptors. Nature 347: 182-4 203 citations
5. Manzoni,O., Prezeau, L., Marin, P., Bockaert, J, et al. Nitric oxide induced
blockade of NMDA receptors Neuron 8 : 653- 662 APR 1992 426 citations
6. Lafon-Cazal M., Pietri S., Culcasi M. et Bockaert J. (1993) NMDA-dependent
superoxide production and neurotoxicity. Nature 364: 535-537. 863 citations
7. Spengler D., Waeber C., Pantaloni C., Holsboer F., Bockaert J., Seeburg P.H. et
Journot L. (1993) Differential signal transduction by five splice variants of the
PACAP receptor. Nature 265: 170-175 938 citations
8. Pin J.P., Joly C., Heinemann S.F. et Bockaert J. (1994) Domains involved in the
specificity of G-protein activation in phospholipase C coupled metabotropic

glutamate receptors. EMBO J. 13: 342-348.204 citations
9. Chavis, P ;, Fagni, L, Lansman,J.B., Bockaert, J (1996) Functional coupling
between ryanodine receptors and L-type calcium channels in neurons Nature 382 :
719-722 194 citations
10. Ango, F, prézeau, L, Muller, T, Worley, P. Pin, J.P., Bockaert, J, Fagni, L (2001)
Agonistindependent activation of mGluRs by intracellular protein Homer. Nature
411 :962-965 164 citations
11. Bécamel C, Alonso G, Galéotti N, Demey E, Jouin P, Ullmer C, Dumuis A,
Bockaert J, Marin P (2002) Synaptic multiprotein complexes associated with
5-HT2c receptors: a proteomic approach. EMBO J 21, 2332-42 70 citations
12. Roussignol G, Ango F, Romorini S, Tu JC, Sala C, Worley PF, Bockaert J, Fagni L
(2005) Shank expres-sion is sufficient to induce functional dendritic spine synapses
in spiny neurons. J Neurosci 25, 3560-70.25
13. Jean A, Conductier G, Manrique C, Bouras C, Berta P, Hen R, Charnay Y,
Bockaert J, Compan V. Anorexia induced by activation of serotonin 5-HT4
receptors is mediated by increases in CART in the nucleus accumbens. Proc Natl
Acad Sci U S A. 2007 104(41):16335-40.
14. Delcourt N, Thouvenot E, Chanrion B, Galéotti N, Jouin P, Bockaert J, Marin P.
PACAP type I receptor transactivation is essential for IGF-1 receptor signalling and
antiapoptotic activity in neurons. EMBO J. 2007 26(6):1542-51. 2007
15. Bertaso F, Zhang C, Scheschonka A, de Bock F, FontanaudP, Marin P, Huganir RL,
Betz H, Bockaert J, Fagni L, Lerner- Natoli M. PICK1 uncoupling from mGluR7a
causes absence-like seizures. Nat Neurosci. 2008 (8):940-8 .
16. Barthet G, Carrat G, Cassier E, Barker B, Gaven F, Pillot M, Framery B, Pellissier
LP, Augier J, Kang DS, Claeysen S, Reiter E, Banères JL, Benovic JL, Marin P,
Bockaert J, Dumuis A. Betaarrestin1 phosphorylation by GRK5 regulates G
proteinindependent 5-HT4 receptor signalling.EMBO J. 2009 Sep
16;28(18):2706-18.
17. Mitri C, Soustelle L, Framery B, Bockaert J, Parmentier ML, Grau Y.Plant
insecticide L-canavanine repels Drosophila via the insect orphan GPCR DmX.
PLoS Biol. 2009 Jun 30;7(6):e1000147.

ABSTRACT
New Modes of GPCR Signalling
Joel Bockaert, Federica Bertaso, Carine Becamel, Maryline Labasque, Mireille
Lerner-Natoli, Laurent Fagni, Philippe.Marin.
CNRS UMR5203,Institut de Génomique Fonctionnelle,Montpellier,France,INSERM,
U661,Montpellier France and University of Montpellier
For a long time, we thought that GPCRs were part of a two-dimentional signaling
pathways. It was believed that ligand-activated GPCRs were only activating one or a
limited number of heterotimeric G proteins . However, since according to Paul Valery
« what is simple is certainly wrong » the control of cell physiology by GPCRs is far
more complex. We will illustrate, with 3 examples, this complexity. The first one
concerns a new type of transactivation between GPCRs and receptor tyrosine kinases
(RTKs). Classically GPCRs are known to be transactivated by RTKs. However, there
are some examples of the reverse situation. Indeed we provide data showing that
IGF-Rs transactivate PACAP-Rs in neurons. Phosphorylation of PACAP-Rs, induced
by IGF1, lead to un-liganded PACAP-Rs activation and their coupling to Gs. The
second one is the key role of GPCR-associated signalling in fine-tuning of their
signalling. mGluR7 negatively control the P/Q channels of pre-synaptic terminals an
effect which require absolutely the physical association of their c-terminal PDZ ligand
with PICK1. We have shown, in vivo, that disconnecting mGluR7 from PICK1 (with
cell-permeant dominant-negative peptide and knock-in mice) causes absence-like
seizures. The third one is the illustration that GPCRs can signal without any G
activation. This will be illustrated here by showing that long-lasting (one hour)
activation of ERK pathway by 5-HT2c-Rs is mediated via a complex formed by the
receptor C-terminal associated and calmodulin and -arrestin.

INVITED SPEAKER’S INFO
Prof. Kun-Liang Guan
PhD, Professor
UCSD Moores Cancer Center,
RM5344, 3855 Health Sciences Dr.,
La Jolla, CA 92093-0815.
Tel: 858-822-7945,
E-mail: kuguan@ucsd.edu
Education
1978-1982 Undergraduate student in the Department of Biology.
Hangzhou University, China, B.S.
1982-1983 CUSBEA student, Graduate student in Shanghai Institute of Plant
Physiology, Shanghai, China
1983-1989 Graduate student in the Department of Biochemistry,
Purdue University, West Lafayette, Indiana, Ph.D. (Dec.17, 89)
1989-1991 Post doctoral fellow in the Department of Biochemistry,
Purdue University (Mentor, Jack Dixon)
Academic Appointments
1992-1996 Assistant Professor, Department of Biological Chemistry,
University of Michigan
1996-2000 Associate professor, Department of Biological Chemistry,
University of Michigan
2000-2007 Professor, Department of Biological Chemistry, University of Michigan
2002- Visiting professor, Department of Chemistry and Biochemistry,
University of Colorado at Boulder
2003-2007 Research professor, Life Sciences Institute, University of Michigan
2003-2007 Halvor N. Christensen Collegiate Professor in Life Sciences,
University of Michigan
2005- Adjunct professor, Fudan University, China
2007- Visiting Research professor, Life Sciences Institute,
University of Michigan
2007- Professor, Department of Pharmacology,
Member, Moores Cancer Center, University of California, San Diego
Selected Publications over 180
1. Guan, K-L., Broyles, S.S., and Dixon, J.E. (1991) Vaccinia virus encodes a Tyr/Ser
protein phosphatase. Nature. 350, 359-362.
2. Zhang, Z., Wang, Y., Vikis, H. G., Anderson, M.W., Sills, R.C., Holliday, W.,
Devereux, T.R., Guan, K-L., and You, M. (2001) Wild type ras can inhibit lung

carcinogenesis in mice. Nature Genetics. 29, 25-33.
3. Inoki, K., Li, Y., Zhu, T., Wu, J. and Guan, K-L. (2002) TSC2 is phosphorylated
and inhibited by AKT and suppresses mTOR signaling. Nature Cell Biol. 4,
648-657. (Selected by Nature Cell Biol as one of the landmark papers).
4. Li, W., Lee, J., Vikis, H., et al, and Guan, K-L. (2004) Activation of FAK and
Src are receptor proximal events required for netrin signaling. Nature.
Neuroscience. 7, 1213-1221.
5. Kim, E., Goraksha, P., Li, L., Neufeld, T.P., and Guan, K-L. (2008) Regulation of
TORC1 by Rag GTPases in nutrient response. Nature Cell Biol. 10, 935-945.
6. Chang, J., Wang, Z., Tang, E., Fan. Z., McCauley, L., Franceschi, R., Guan, K-L.,
Kresbach, PH., and Wang, C-Y. (2009) Inhibition of Osteoblast functions by
IKK/NFkB in osteoporosis. Nature.Med. 15, 682-9.
7. Inoki, K., Corradetti, and Guan, K-L. (2005) Disease associated with TOR
Dysregulation. Nature Genetics. 37, 19-24.
8. Guan, K-L. and Dixon, J.E. (1990) Protein tyrosine phosphatase activity of an
essential virulence determinant in Yersinia. Science. 249, 553-556.
9. Simon, A. R., Vikis, H.G., Fanburg, B.L., Stewart, S., Cochran, B.H., and Guan,
K.-L. (2000) Regulation of STAT3 by direct binding to the small G protein Rac1.
Science. 290, 144-147.
10. Zhao, S., Lin, Y., Xu., W., Jiang, W., Zha, Z., Wang, P., Yu, W., Li, Z., Gong, L.,
Peng, Y., Ding, J., Lei, Q., Guan, K-L., Xiong, Y. (2009) Glioma-derived
Mutations in IDH1 Dominantly Inhibit IDH1 Catalytic Activity and Induce HIF-1.
Science. 324, 261-5.
11. Zhao, S., Xu, W., Jiang, W., Yu, W., Lin, Y., Zhang, T., Yao, J., Zeng, Y., Li, H.,
Li, Y., Shi., J., An, W., Hancock, S.M., He., F., Qin, L., Chin, J., Yang, P., Chen,
X., Lei, Q., Xiong, Y., Guan, K-L., (2010) Regulation of cellular metabolism by
protein lysine acetylation. Science. 327, 1000-1004.
12. Wang, Q., Zhang, Y., Yang, C., Xiong, H., Lin, Y., Yao, J., Li, H., Xie, L., Zhao,
W., Yao, Y., Ning, Z-B., Zeng, R., Xiong, Y., Guan, K-L., Zhao, S., Zhao, G-P.,
(2010) Acetylation of metabolic enzymes coordinates carbon source utilization and
metabolic flux. Science. 327, 1004-1007.
13. Inoki, K., Zhu, T., and Guan, K-L. (2003) TSC2 mediates cellular energy response
to control cell growth and survival. Cell. 115, 577-590.
14. Inoki, K., Ouyang, H., Zhu, T., Lindvall, C., Wang, Y., Yang, Q., Bennett, C.,
Harada, Y., Stankunas, K., Wang, C., He, H., MacDougald, O., You, M., Williams,
B., and Guan, K-L. (2006) TSC2 integrates Wnt and cellular energy signals through
a coordinated phosphorylation by AMPK and GSK3 to regulate cell growth. Cell.
126, 955-68.
15. Mori, H., Inoki, K., Münzberg, H., Opland, D., Faouzi, M., Villanueva, E.C.,
Ikenoue, T., Kwiatkowski, D., MacDougald, O.A., Myers, M.G. Jr., and Guan, K-L.
(2009) Critical role for hypothalamic mTOR activity in energy balance. Cell
Metabolism. 9, 362-74.
16. Zhao, B., Ye, X., Yu, J., Li, L., Li, W., Li, S., Lin, JD., Wang, C- Y., Chinnaiyan,
AM., Lai, Z-C., and Guan. K-L. (2008) TEAD mediates YAP dependent gene
induction and growth control. Genes & Development. 22, 1962-71.

ABSTRACT
Regulation of Cellular Metabolism by Protein Lysine Acetylation
Shimin Zhao, Wei Xu , Wenqing Jiang, Wei Yu, Yan Lin, Tengfei Zhang, Qunying Lei,
Yue Xiong and Kun-Liang Guan
Molecular and Cell Biology Laboratory, Institute of Biomedical Sciences, Fudan
University, Shanghai 20032, China
Protein lysine acetylation has emerged as a key posttranslational modification in cellular
regulation, in particular through modification of histones and nuclear transcription
regulators. We show that lysine acetylation is a prevalent modification in enzymes that
catalyze intermediate metabolism. Virtually every enzyme in glycolysis,
gluconeogenesis, tricarboxylic acid (TCA) cycle, urea cycle, fatty acid metabolism, and
glycogen metabolism was acetylated in human liver tissue. The concentration of
metabolic fuels, such as glucose, amino acids, and fatty acids, influenced acetylation
status of metabolic enzymes. Acetylation activated enoyl-Coenzyme A
hydratase/3-hydroxyacyl-Coenzyme A dehydrogenase (EHHADH) in fatty acid
oxidation and malate dehydrogenase (MDH) in the TCA cycle, inhibited
argininosuccinate lyase (ASL) in the urea cycle, and destabilized phosphoenolpyruvate
carboxykinase (PEPCK) in gluconeogenesis. Our study reveals that acetylation plays
a major role in metabolic regulation.

INVITED SPEAKER’S INFO
Prof. Jean-Philippe Pin
PhD, Directeur de Recherche (DR1) CNRS
Inst. for Functional Genomics,
CNRS–INSERM –UM1–UM2
Dept of Molecular Pharmacology
141 rue de la Cardonille 34094
Montpellier cedex 5–France
Phone: +33 467 14 2988
FAX: +33 467 54 2996
Email: jppin@igf.cnrs.fr
Education
1987 Diplôme de doctorat (Ph. D.),
University of Montpellier.
Centre CNRS-INSERM de Pharmacologie- Endocrinologie.
1984 Diplôme d'Etudes Approfondies (DEA) of Molecular Biology
University of Montpellier.
Centre CNRS-INSERM de Pharmacologie- Endocrinologie
1983 Master in Physiology- Certificate of Biochemistry
Université Pierre et Marie Curie - Paris VI.
1981 Ecole Normale Supérieure de Paris (45, rue d'Ulm)
Selected Publications
1. Sladeczek F, Pin J-P, Récasens M, Bockaert J and Weiss S (1985) Glutamate
stimulates inositol phosphate formation in striatal neurones. Nature. 317: 717-719.
2. Weiss, S., Pin, J.-P., Sebben, M., Kemp, D., Sladeczek, F., Gabrion, J. and
Bockaert, J. (1986) Synaptogenesis of cultured striatal neurones in serum-free
medium: a morphological and biochemical study. Proc. Natl. Acad. Sci. (USA),
83:2238-2242.
3. Dumuis, A., Sebben, M., Haynes, L., Pin, J.-P. and Bockaert, J. (1988) NMDA
receptors activate the arachidonic acid cascade system in striatal neurons. Nature.
336:68-70.
4. Dumuis A, Pin J-P, Oomagari K, Sebben M and Bockaert J (1990) Arachidonic
acid released from striatal neurons by joint stimulation of ionotropic and
metabotropic quisqualate receptors. Nature. 347: 182-184.
5. Rassendren F A, Lory P, Pin J-P and Nargeot J (1990) Zinc has opposite effects on
NMDA and non-NMDA responses in Xenopus oocytes injected with rat brain RNA.
Neuron, 4:733-740.
6. Pin J-P, Waeber C, Prézeau L, Bockaert J and Heinemann S F (1992) Alternative
splicing generates metabotropic glutamate receptors inducing different patterns of
calcium release in Xenopus oocytes. Proc. Natl. Acad. Sci. (USA),
89:10331-10335.
7. Pin J-P, Joly C, Heinemann SF and Bockaert J (1994) Domains involved in the

specificity of G protein activation in phospholipase C coupled metabotropic
glutamate receptor. EMBO J 13: 342-348.
8. Ango F, Prézeau L, Muller T, Worley PF, Pin J-P, Bockaert J and Fagni L (2001)
Agonist-independent activation of mGluRs by the intracellular interacting protein,
Homer. Nature 411: 962-965.
9. Galvez T, Duthey B, Kniazeff J, Blahos J, Rovelli G, Bettler B, Prézeau L and Pin
J-P (2001) Allosteric interactions between GB1 and GB2 subunits are required for
optimal GABAB receptor function. EMBO J 20: 2152-2159.
10. Bessis A-S, Rondard P, Gaven F, Brabet I, Triballeau N, Prézeau L, Acher F and
Pin J-P (2002) Closure of the Venus Flytrap module of family 3 GPCRs is required
for their activation: insights from mutations converting antagonists into agonists.
Proc. Natl. Acad. Sci (USA), 99:11097-11102.
11. Kniazeff J, Bessis A.-S, Maurel D, Ansanay H, Prezeau L and Pin J-P (2004)
Closed state of both binding domains of homodimeric mGlu receptors is required
for full activity. Nat. Str. Mol. Biol., 11: 706-713.
12. Goudet C, Gaven F, Kniazeff J, Vol C, Liu J, Cohen-Gonsaud M, Acher F, Prézeau
L and Pin J-P (2004) Heptahelical domain of metabotropic glutamate receptor 5
behaves like rhodopsin-like receptors. Proc. Natl. Acad. Sci (USA), 101, 378-383.
13. Hlavackova V, Goudet C, Kniazeff J, Zikova A, Maurel D, Vol C, Trojanova J,
Prézeau L, Pin J-P, Blahos J (2005) Evidence for a single heptahelical domain
being turned on upon activation of a dimeric GPCR. EMBO J. 24:499-509.
14. Damian M, Martin A, Mesnier D, Pin JP, Banères JL (2006) Asymmetric
conformational changes in a GPCR dimer controlled by G-proteins. EMBO J.
25:5693-5702.
15. Rondard P, Huang S, Monnier C, Tu H, Blanchard B, Oueslati N, Malhaire F, Li Y,
Maurel D, Trinquet E, Labesse G, Pin* J-P, Liu J (2008) Functioning of the dimeric
GABAB receptor extracellular domain revealed by glycan wedge scanning. EMBO
J 27:1321-1332.
16. Maurel D, Comps-Agrar L, Brock C, Rives M-L, Bourrier E, Ayoub MA, Bazin H,
Tinel N, Durroux T, Prézeau L, Trinquet E, Pin J-P (2008) Cell surface
protein-protein interaction analysis with combined time-resolved FRET and
snap-tag technologies: application to GPCR oligomerization. Nat Meth 5:561-567.
17. Rives ML, Vol C, Fukazawa Y, Tinel N, Trinquet E, Ayoub MA, Shigemoto R, Pin
JP, Prezeau L (2009) Crosstalk between GABA(B) and mGlu1a receptors reveals
new insight into GPCR signal integration. EMBO J. 28:2195-2208.
18. Bartfai T, Benovic J, Bockaert J, Bond R, Bouvier M, Christopoulos A, Civelli O,
Devi L, George S, Inui A, Kobilka B, Leurs R, Neubig R, Pin J-P, Quirion R,
Roques B, Sakmar T, Seifert R, Stenkamp R, Strange P (2004) Twenty Questions
on GPCRs. Nat Rev Drug Discovery. 3:577-626.

ABSTRACT
Sensing mGlu Receptor Activation
Jean-Philippe Pin 1, Etienne Doumazane 1, Pauline Scholler 1, Siluo Huang 2, Eric
Trinquet 3, Jianfeng Liu 2 & Philippe Rondard 1
1 CNRS UMR5203, INSERM U661, University of Montpellier, Department of Molecular
Pharmacology, Institute of Functional Genomics, Montpellier, France. 2 Huazhong
University of Science and Technology, Wuhan, China, 3 CisBio International,
Bagnols/Cèze , France
Metabotropic glutamate receptors play key role in the modulation of many synapses,
acting either on the post-synaptic element from where they control post-synaptic
ionotropic receptor activity, or on pre-synaptic elements where they control
neurotransmiter release. Eight genes encoding mGluRs have been identified in
vertebrate genomes, been classified into 3 groups. Group-I mGluRs (mgluR1 and
mGluR5) are post-synaptic receptors coupled to Gq type of G-proteins, therefore
activating phospholipase C, whereas group-II (mGluR2 and mGluR3) and group-III
(mGluR4, 6, 7 and 8) are mostly pre-synaptic receptors coupled to Gi/o types of
G-proteins. These receptors then, represent new promising targets for he development
of new drugs for the treatment of many neurologic and psychiatric diseases.
These receptors are constitutive dimers, each subunit being made of three major
domains: i) the venus flytrap domain (VFT) where glutamate bind; ii) a cystein-rich
domain (CRD); and iii) the 7 transmembrane domain (7TM) responsible for G-protein
activation. How agonist binding leads to the change in conformation in the 7
transmembrane domain remains unclear. The first crystal structures of the mGluR1 VFT
with or without bound agonist revealed two major differences: 1) a closed state of the
agonist occupied VFT; and 2) a change in the relative position of the two VFTs within
the dimeric structure. It was then postulated that the relative movement between the
VFTs constitutes a key step in receptor activation. However, new crystal structures did
not confirm this change in the relative position. Using cell surface labeling of each
subunit with FRET compatible fluorophores, we have been able to monitor, in living
cells, any possible movement between the subunits during receptor activation. Such
assay is very sensitive,simple, and can be used to characterize different ligands acting
on mGluRs: agonists, antagonists, positive or negative allosteric modulators. We even
documented the mechanism of action of partial agonists.
Our data are consistent with the initial hypothesis for receptor activation. This was
further demonstrated using receptor mutants that either prevent the relative movement
of the subunits, or mutants in which the two subunits are locked in the active orientation.
These provide important new information on the functioning of these receptors, then
offering new possibilities to develop new types of drugs modulating their activity.

INVITED SPEAKER’S INFO
Prof. Philippe Rondard
PhD, Senior researcher,
Institute for Functional Genomic
Department of Molecular Pharmacology
CNRS UMR5203 – INSERM U661 – Universités
Montpellier 1 & 2, 141, rue de la Cardonille
34094 Montpellier Cedex 5 France
Tel: 33 (0)4 67 14 2912
Fax: 33 (0)4 67 54 2432
Email: philippe.rondard@igf.cnrs.fr
Education
1993-1998 Ph.D in Biochemistry.
University of Paris VII and Institut Pasteur, Paris, France.
1992-1993 Diplôme d'Etudes Approfondies (DEA)
University of Paris VII, France.
Professional Experience
1998-2001 Post-doctoral training in the laboratory of Dr. Henry BOURNE,
University of California San Francisco (UCSF), San Francisco, USA.
1993-1998 Doctoral training in the laboratory of Dr. Hugues BEDOUELLE,
Institut Pasteur, Paris, France.
1995-1996 Military Service as Scientific researcher,
Ecole Normale Supérieure and Institut Curie, Paris.
Selected Publications
1. Monnier C., Dodé C., Fabre L., Teixeira L., Labesse G., Pin J.-P., Hardelin J.-P.
and Rondard P. (2009) PROKR2 missense mutations associated with Kallmann
syndrome impair receptor signalling-activity. Hum. Mol. Genet. 18:75-81.
2. Ahier A., Rondard P., Gouignard N., Khayath N., Huang S., Trolet J., Donoghue
DJ, Gauthier M., Pin J.-P. and Dissous C. (2009) A new family of receptor tyrosine
kinases with a venus flytrap binding domain in insects and other invertebrates
activated by aminoacids. PLoS One 4(5):e5651.
3. Rondard P., Huang S., Monnier C., Tu H., Blanchard B., Oueslati N., Malhaire, F.,
Oueslati, N., Li Y., Trinquet E., Labesse G., Pin J.-P. and Liu J. (2008) Functioning
of the dimeric GABAB receptor extracellular domain revealed by glycan wedge
scanning EMBO J. 27, 1321-1332.
4. Tu. H.*, Rondard P.*, Xu C., Bertaso F., Cao F., Zhang X., Pin J.-P. and Liu J.
(2007) Dominant role of GABAB2 and Gαfor GABAB receptor
mediated-ERK1/2/CREB pathway in cerebellar neurons. Cell signal.19:1996-2002.
* Equally contributed
5. Rondard P., Liu J., Huang S., Malhaire F., Vol C., Pinault A., Labesse, G. and Pin
J.-P. (2006) Coupling of agonist binding to effector domain activation in

metabotropic glutamate-like receptors. J. Biol. Chem. 281, 24653-24661.
6. Liu J., Maurel D., Etzol S., Brabet I., Ansanay H., Pin J.-P. and Rondard P. (2004)
Molecular determinants involved in the allosteric control of agonist affinity in
GABAB receptor by the GABAB2 subunit. J. Biol. Chem. 279, 15824-15830.
7. Pin J.-P., Kniazeff J., Binet V., Liu J., Maurel D., Galvez T., Duthey B.,
Havlickova M., Blahos J., Prézeau L. and Rondard P. (2004). Activation
mechanism of the heterodimeric GABAB receptor. Biochem. Pharmacol. 68,
1565-1572.
8. Pin J.-P., Kniazeff J., Goudet C., Bessis A.-S., Liu J., Galvez T., Acher F., Rondard
P. and Prézeau L. (2004) The activation mechanism of class-III G-protein coupled
receptors (2004). Biol. Cell 96, 335-342.
9. Bessis A.-S., Rondard P., Gaven F., Brabet I., Triballeau N., Prézeau L., Acher F.
and Pin J.-P. (2002) Closure of the Venus flytrap module of mGlu8 receptor and the
activation process: Insights from mutations converting antagonists into agonists.
Proc. Natl. Acad. Sci. U. S. A. 99, 11097-11102
10. Rondard P., Iiri T., Srinivasan S., Meng E., Fujita T. and Bourne HR. (2001).
Mutant G protein �subunit activated by G : a model for receptor activation ?
Proc. Natl. Acad. Sci. U. S. A. 98, 6150-6155.
11. Rondard P. and Bedouelle H. (2000) Mutational scanning of a hairpin loop in the
tryptophan synthase subunit, implicated
in allostery and substrate channeling.
Biol. Chem. 381, 1185-1193.
12. Rondard P. and Bedouelle H. (1998) A mutational approach shows similar
mechanisms of recognition for the isolated and integrated versions of a protein
epitope. J. Biol. Chem. 273, 34753-34759.
13. Rondard P., Brégégère F., Lecroisey A., Delepierre M. and Bedouelle H. (1997a)
Conformational and functional properties of an undecapeptide epitope fused with
the C-terminal end of the maltose binding protein. Biochemistry 36, 8954-8961.
14. Rondard P., Goldberg M.E. and Bedouelle H. (1997b) Mutational analysis of an
antigenic peptide shows recognition in a loop conformation. Biochemistry 36,
8962-8968.

ABSTRACT
mGlu Receptors : Complex Allosteric Machines to Tune Up Synaptic
Transmission
Philippe Rondard1, Etienne Doumazane1, Pauline Scholler1, Eric Trinquet2, Sébastien
Granier1 & Jean-Philippe Pin1
1 CNRS UMR5203, INSERM U661, University of Montpellier, Department of Molecular
Pharmacology, Institute of Functional Genomics, Montpellier, France.
2 CisBio International, Bagnols/Cèze , France
The G-protein coupled receptors activated by the neurotransmitter glutamate (mGluRs)
are made up of two subunits covalently linked by a disulfide bridge. Each protomer
comprises an extracellular domain that binds agonists and a transmembrane heptahelical
domain responsible for G-protein activation. The general organization of mGluRs at the
cell surface, whether they are limited to dimers, or organized into high-order oligomers,
as well as the functioning of the dimeric receptors remain unclear.
Here, we first examined whether mGluRs can assemble into heteromeric complexes. To
that aim we describe a new approach enabling the specific labeling of two cell surface
proteins carrying SNAP- or CLIP-tags, with two distinct fluorophores compatible with
time-resolved FRET. This approach allowed us to quantify both homomeric and
heteromeric populations. Our data revealed that some, but not all pairs of mGluRs can
indeed form heteromeric entities, and this is further supported by biochemical analysis
and functional complementation studies. By FRET competition and saturation analysis,
we show that these complexes are limited to heterodimers. In addition to describing a
new way to analyze cell surface receptor complexes, our data reveal a new possible
level of complexity within the mGluR family.
Second, whereas dimerization of the extracellular domains is essential for the activation,
it is not known if the function of the transmembrane domains also depends on
dimerization. To address this question, we analyzed the G protein coupling of purified
mGluR membrane domain reconstituted into phospholipid bilayer nanodisc with a
controlled stoichiometry: one or two protomers per disc. Nanodiscs containing only one
mGluR membrane domain are able to activate G proteins in response to a positive
allosteric modulator, demonstrating that monomeric mGluR membrane domain can fold
and function independently from the rest of the receptor. These results suggest that
dimer requirement in mGluR function originate from an obligate communication
between the extracellular and transmembrane domains in the receptor for activation by
glutamate, rather than a necessity for specific association between the two membrane
domains.

INVITED SPEAKER’S INFO
Prof. Denise Montell
PhD, Professor, Director,
Department of Biological Chemistry,
Center for Cell Dynamics
Johns Hopkins School of Medicine,
855 North Wolfe Street, Baltimore, MD 21205-2185
Phone: (410) 614-2016
Fax: (410) 614-8375
Email: dmontell@jhmi.edu
Education and Training
1979-1983 B. A.,University of California, San Diego, Biochemistry and Cell Biology
1983-1988 Ph.D., Stanford University, Neurosciences
1988-1990 Postdoctoral studies, , Carnegie Institution, Developmental Genetics
Professional Experience
1990-1992 Staff Associate, Carnegie Institution, Department of Embryology
1992-1998 Assistant Professor, Department of Biological Chemistry
Johns Hopkins University School of Medicine
1998-2002 Associate Professor, Department of Biological Chemistry
Johns Hopkins University School of Medicine
2002- Professor, Department of Biological Chemistr
Johns Hopkins University School of Medicine
1999- Director, Graduate Program in Biological Chemistry
2006- Director, Center for Cell Dynamics.
Selected Publications
1. Geisbrecht, E. and Montell, D. J. (2002) Requirement for Myosin VI in
E-cadherin-mediated border cell migration, Nature Cell Biol 4, 616-620. Cover
photo.
2. Montell, D. J. (2003) Border cell migration: the race is on. Nature Reviews
Molecular Cell Biology 4, 13-24.
3. Naora, H. and Montell, DJ. (2005) Ovarian cancer metastasis: Integrating insights
from disparate model organisms. Nature Reviews Cancer, 5, 355-66.
4. Jang, A. C.-C., Chang, Y.-C., Bai, J. and Montell, D.J. (2009) Border cell migration
requires integration of spatial and temporal signals by the BTB protein Abrupt.
Nature Cell Biol 11:569-79.
5. Wang, X., He, L., Wu, Y., Hahn, K. and Montell, D. J. Light-mediated activation
reveals a key role for Rac in collective guidance of cell movement in vivo, Nature
Cell Biol, in press.
6. Montell, D. J., Keshishian, H., and Spradling, A. C. (1991) Laser ablation studies of
the role of the Drosophila oocyte nucleus in pattern formation. Science 254,

290-293
7. Montell, D. J. (2008) Morphogenetic Cell Movements: Diversity from
Combinatorial Use of Modular Mechanical Properties. Science, 322:1502-5.
8. Johnson, D. M., Newby, R. F. and Bourgeois, S. (1984) Membrane permeability as
a determinant of dexamethasone resistance in murine thymoma cells.Cancer
Research 44, 2435-2440.
9. Montell, D. J. and Goodman, C. S. (1988) Drosophila substrate adhesion molecule:
sequence of laminin B1 chain reveals domains of homology with mouse. Cell 53,
463- 473.
10. Montell, D. J., Rørth, P. and Spradling, A. C. (1992) slow border cells, a locus
required for a developmentally regulated cell migration during oogenesis encodes
Drosophila C/EBP. Cell 71, 51-62.
11. Murphy, A. M., Lee, T., Andrews, C., Shilo, B.-Z. and Montell, D. J. (1995) The
Breathless FGF receptor homolog, a downstream target of Drosophila C/EBP in the
developmental control of cell migration. Development 121, 2255-2263.
12. Lee, T., Feig, L. and Montell, D. J. (1996) Two distinct roles for Ras in a
developmentally regulated cell migration. Development 122, 409-418.
13. Liu, Y. and Montell, D. J. (1999) Identification of Mutations that Cause Cell
Migration Defects in Mosaic Clones. Development 126, 1869-1878.
14. Montell, D. J. (1999) The Genetics of Cell Migration in Drosophila melanogaster
and Caenorhabditis elegans Development. Development 126, 3035-3046
15. Bai, J., Uehara, Y. and Montell, D. J. (2000) Regulation of invasive cell behavior
by Taiman, a Drosophila protein related to AIB1, a steroid receptor coactivator
amplified in breast cancer. Cell 103, 1047-1058.
16. Liu, Y. and Montell, D. J. (2001) Jing, a downstream target of slbo required for
developmental control of border cell migration. Development 128, 321-330.
17. Silver, D. and Montell, D. J. (2001) Paracrine Signaling through the JAK/STAT
Pathway Activates Invasive Behavior of Ovarian Epithelial Cells in Drosophila,
Cell 107, 831-841
18. Yoshida, H., Cheng, W., Hung, J., Montell, D., Geisbrecht, E., Rosen, D., Liu, J. and Naora, H.
(2004) Lessons from border cell migration in the Drosophila ovary:A role for myosin VI in
migration of human ovarian cancer cells, Proc. Natl. Acad. Sci. (USA) 101,
8144-8149.
19. Montell, D. J. (2006) A Kinase Gets Caspases into Shape. Cell 126, 450-2.
20. Silver, D. L., Naora H., Liu, J., Cheng, W. and Montell, D. J. (2004) Activated
STAT3: localization in focal adhesions and function in ovarian cancer cell motility.
Cancer Research 15, 3550-8.

ABSTRACT
Manipulation of Rac Activity and Cell Motility in vivo Using Light
Xiaobo Wang, Li He, Yi Wu, Klaus Hahn and Denise J. Montell
Department of Biological Chemistry, Center for Cell Dynamics Johns Hopkins School
of Medicine, Baltimore, USA
The small GTPase Rac induces actin polymerization, membrane ruffling and focal
contact formation in cultured single cells, can promote growth cone guidance, dynamic
dendrite growth and has been implicated in learning and memory. Recently,
photoactivatable analogues of Rac (PA-Rac) have been developed, allowing rapid and
reversible activation or inactivation of Rac using light. In cultured single cells,
light-activated Rac leads to focal membrane ruffling, protrusion and migration. Here we
report that focal activation of Rac is effective in vivo. Light-mediated activation of
Rac in one part of one cell caused local ruffling and protrusion. Moreover, activation
or inactivation of Rac in one cell of a cluster caused a dramatic response in the other
cells, and set the direction of movement for the whole group. These results suggest
that the cells sense direction as a group according to relative levels of Rac activity.
Communication between cells of the cluster required Jun N-terminal kinase (JNK) but
not guidance receptor signalling. These studies further suggest that photoactivatable
proteins are likely to be a new and generally effective class of tools for manipulating
local protein activities in vivo.

INVITED SPEAKER’S INFO
Prof. Jianfeng Liu
PhD, Professor, Vice director,
Key laboratory of molecular biophysics of ministry of
education
Huazhong University of Science and Technology (HUST)
1037, Luoyu Road, Hubei, Wuhan, 430074, P.R.China
Tel : 0086-27-87792031
Fax : 0086-27-87792024
Email : jfliu@mail.hust.edu.cn
Education
1994 -1999 Ph.D. Molecular and Cell Biology,
University of Paris XII, Paris, France.
1993 -1994 M.S. Biochemistry and Biotechnology,
University of Paris XIII, Paris, France.
1989 -1993 B.S. Biochemistry, Department of Biochemistry,
Wuhan University, Wuhan, China
Research Experience
2002- Professor, College of Life Science and Technology,
HuaZhong University of Science and technology, Wuhan, China.
2001-2002 Staff Scientist in Aventis Pharma, Bridgewater, New Jersey, USA.
1999-2001 Post-doc Fellow
Institut Cochin Génétique Moléculaire (ICGM), Paris, France.
1994-1999 Research Assistant in the Laboratoire de Recherche sur la Croissance
Cellulaire, la Réparation et la Régénération Tissulaire,
University of Paris XII, Creteil, France.
1993-1994 Research assistant, Institut d’oncologie Cellulaire et Moléculaire
Humaine,University of Paris XIII, Bobigny, France.
Selected Publications
1. Tu HJ, Xu CJ, Zhang WH, Liu QY, Rondard P, Pin JP, Liu JF. (2010) GABAB
receptor activation protects neurons from apoptosis via IGF-I receptor
transactivation. J. Neurosci., 30(2): 749-759.
2. Cao JH, Huang SL, Qian J, Huang JL, Jin L, Su ZX, Yang J, Liu JF. (2009)
Evolution of the class C GPCR Venus flytrap modules involved positive selected
functional divergence. BMC Evol Biol., 9:67 .
3. Rondard P, Huang SL, Monnier C, Tu HJ, Blanchard B, Oueslati N, Malhaire F,
Maurel D, Trinquet E, Li Y, Labesse G, Pin JP, Liu JF (2008). Functioning of the
dimeric GABAB receptor extracellular domain revealed by glycan wedge scanning.
EMBO J, 27(9):1321-32
4. Li X, Cao JH, Li Y, Rondard P, Zhang Y, Yi P, Liu JF, Nan FJ. (2008)

Activity-based Probe for Specific Photo-affinity Labelling of GABAB Receptors in
Living Cells: Design, Synthesis and Biological Evaluation. J. Med. Chem
12;51(11):3057-60.
5. Huang S., Liu JF., Labesse G., Pin J.-P. & Rondard P. Rearrangement of the
cystein-rich domains during metabotropic glutamate receptors activation. Manuscrit
in preparation
6. Tu HJ, Rondard P, Xu CJ, Bertaso F, Cao FN, Zhang XY, Pin JP, Liu JF. (2007)
GABAB2 subunit and Gßγ are necessary for GABAB receptor mediated-ERK1/2
phosphorylation in cerebellar granule neurons. Cell. Signal., 19(9):1996-2002.
7. Zhang YL, Zhang HQ, Hua SN, Ma LH, Chen C, Liu XY, Jiang LQ, Yang HM,
Zhang PC, Yu DQ, Guo YL, Tan XH, Liu JF. (2007) Identify two herbal
compounds with potential cholesterol-lowing activity. Biochem. Pharmaco.,
74(6):940-7.
8. Rondard P, Liu JF, Huang SL, Malhaire F, Vol C, Pinault A, Pin JP. (2006)
Coupling of agonist binding to effector domain activation in metabotropic
glutamate-like receptors. J. Biol. Chem., 281(34):24653-61. (co-first and
corresponding author)
9. Pin JP, Kniazeff J, Liu JF, Binet V, Goudet C, Rondard P, Prézeau L. (2005)
Allosteric functioning of dimeric class C G-protein-coupled receptors. FEBS
Journal., 272(12): 2947-2955.
10. Liu JF, Maurel D, Etzol S, Brabet I, Pin JP, Rondard P.(2004) Molecular
determinants involved in the allosteric control of agonist affinity in GABAB
receptor by the GABAB2 subunit. J. Biol. Chem. , 279(16):15824-30.
11. Goudet C, Gaven F, Kniazeff J, Vol C, Liu JF, Cohen-Gonsaud M, Acher F,
Prezeau L, Pin JP. (2004) Heptahelical domain of metabotropic glutamate
receptor 5 functions like rhodopsin-like receptors. Proc. Natl. Acad. Sci., 101(1):
378-383.
12. Liu JF, Crepin M, Liu JM, Barritault D, Ledoux D. (2002) FGF-2 and TPA induce
matrix metalloproteinase-9 secretion in MCF-7 cells through PKC activation of the
Ras/ERK pathway. Biochem. Biophys. Res. Com., 293: 1174-1182.
13. Liu JF, Chevet E, Lemaitre G, Barritault D, Larose L, Crepin M. (1999)
Functional Rac-1 and Nck signaling networks are required for FGF-2-induced DNA
synthesis in MCF-7 cells. Oncogene., 18: 6425-6433.
14. Liu JF, Issad T, Chevet E, Ledoux D, Courty J, Caruelle JP, Barritault D; Crepin M,
Bertin B. (1998) Fibroblast growth factor-2 has opposite effects on human breast
cancer MCF-7 cell growth depending on the activation level of the
mitogen-activated protein kinase pathway. Eur. J. Biochem., 258: 271-276.
15. Liu JF, Bagheri-Yarmand R, Xia YL, Crepin M. (1997) Modulations of breast
fibroblast and carcinoma cell interactions by a dextran derivative (CMDB7).
Anticancer Res., 17: 253-258.

ABSTRACT
Activation Mechanism of GABAB Receptor
Chanjuan Xu, Wenhua Zhang, Xin Lin, Ming Jiang, Haijun Tu, Siluo Huang, Jianfeng
Liu*
Sino-France Laboratory for Drug Screening, Key Laboratory of Molecular Biophysics of
Ministry of Education, School of Life Science and Technology, Huazhong University of
Science and Technology, WuHan, 430074
Class-C G-protein coupled receptors (GPCRs) represent a distant group among the large
family of GPCRs. This class includes the receptors for the main neurotransmitters
including glutamate and γ-aminobutyric acid (GABA), and the receptors for Ca 2+ , some
taste and pheromone molecules, as well as some orphan receptors. Class-C GPCRs
possess a heptahelical domain (HD) involved in heterotrimeric G-protein activation, but
most of them also have a large extracellular Venus Fly Trap (VFT) domain responsible
for agonist recognition and binding. Class -C GPCRs are dimers, either homo or
hetero-dimers. GABAB receptor is consisted of GABAB1 and GABAB2 subunits and
was the first heteromeric GPCR identified. The VFT domain of GABAB1 subunit
contains binding sites for neurotransmitter, agonists or antagonists, while the HD
domain of GABAB2 subunit is responsible for G-protein coupling. Therefore, both
subunits are necessary for a functional GABAB receptor. In this study, we demonstrated
the specific role of each subunit in ligand recognition, intramolecular transduction,
G-protein activation, and allosteric modulation. We also showed that the specific
activation of GABAB receptor leads to G-protein activation, which in turn activate
downstream signaling such as MAPK and PI3K/Akt pathway through trans-activation
of a receptor tyrosine kinase (RTK). To the best of our knowledge, this is the time to
report the trans-activation of a RTK by GABAB receptor.

INVITED SPEAKER’S INFO
Prof. Yong Shen
PhD, Senior scientist/Professor
Haldeman Laboratory of Molecular and Cellular
Neurobiology
Sun Health Research Institute
10515 West Santa Fe Drive, Sun City, AZ, 85351, U.S.A.
Phone: (623)876-5456
Fax: (623)876-6688
Email: Yong.Shen@bannerhealth.com
Education
1983 Nanjing University, China BSc. Physiology
1986 Shanghai Inst. of Physiology, Academy of Sciences MSc. Neurophysiology
1989 State University of New York (SUNY) Ph.D. Neuroscience
Selected Publications
1. Ewers M, Cheng X, Nural HF, Walsh C, Meindl T, Teipel SJ, Buerger K, Shen Y,
Hampel H. (2010). CSF- BACE1 activity predicts hippocampus atrophy in
Alzheimer’s disease. J Alzheimers Dis, Apr 26.
2. Cheng X., Yang L., He P, Li R., Shen Y. (2010). Differential activation of TNF
receptors distinguishes Alzheimer from normal brains. J Alzheimers Dis,
19(2):621-30.
3. Ghosal P., Vogt D, Man L, Shen Y., Lamb B., Pimplikar, SW (2009). Alzheimer's
disease-like pathological features in transgenic mice expressing the APP
intracellular domain. Proc Natl Acad Sci U S A. 106(43): 18367-18377.
4. He, P., Staufenbiel M and Shen, Y. (2008). Interruption of β-catenin signaling
reduces neurogenesis in Alzheimer’s disease’s brains. Nature Proceedings,
September, and J.Neurosci. (2009), 29(20):6545-57.
5. The paper is selected as World Faculty 1000 and Feature Article Xia W., Yang T.,
Imelda M. Smith I. M., Shen Y., Walsh D. M. and Dennis J. Selkoe, D. J. (2009). A
specific ELISA for measuring amyloid β-protein oligomers in human plasma and
the brains of Alzheimer patients. Archives of Neurology, 66(2):190-9.
6. Nural F., He P, Beach T, Sue L, Xia W, and ShenY. (2009). Disassembled DJ-1
high molecular weight complex in cortex in cortex mitochondria from Parkinson’s
disease Patients. Molecular Neurodegeneration.
7. Kozikowski, A. P., Chen, Y., Subahasish, T., Zhong, Z., Melissa A. D’Annibale,
M.A., Shen, Y. and Langley, B.C. (2010). PKC activation and HDAC inhibition -A
dual drug approach to Alzheimer’s disease that reduces Aβ production while
blocking oxidative stress. Proc Natl Acad Sci U S A.. Submitted.
8. Chirapu, S. R, Pachaiyappan, B, Zhong, Z., Abdul-Hay, S. O., Yuan, H., Thatcher,
G., Shen, Y., Kozikowski, A. P, Petukhov, P. A. (2008). Molecular modeling,
Synthesis and Activity Studies of Inhibitors Targeting Alzheimer’s disease

β-Secretase (BACE1). Biological Medicinal Chemistry Letters, 19(1): 264-74.
9. Ewers, M., Zhong, Z., Teipel, S., Büger, K., Wallin, A., Blennow, K., and Shen, Y*.
and Hampel, H*.(2008). Increased CSF-BACE1 activity is associated with
ApoE-ε4 genotype in subjects with mild cognitive impairment and Alzheimer’s
disease, Brain, 131:1252-8.
10. Zhong, Z., Ewers, M., Teipel, S., Büger, K., Wallin, A., Blennow, K., Hampel, H.,
and Shen, Y. (2007). High Levels of Beta-Secretase (BACE1) in Cerebrospinal
Fluid as a Predictor of Risk in Mild Cognitive Impairment, the early stage of
Alzheimer ’s disease, Archive of General Psychiatry, 64(6):718-26.
11. He P, Zhong Z, Lindholm K, Berning L, Lee W, Lemere C, Staufenbiel M, Li R
and ShenY. (2007). Deletion of TNF death receptor inhibits amyloid beta
generation and prevents learning and memory deficits in Alzheimer’s mice.
Journal of Cell Biology, 178(5): 829-841. Highlighted as “Death receptor takes
central stage” by Nature Neuroscience,October, 2007; Hot story by Science, 2007.
12. Tesco G, Koh YH, Kang EL, Cameron AN, Das S, Sena-Esteves M, Hiltunen M,
Yang SH, Zhong Z, Shen Y, Simpkins JW, Tanzi RE. (2007). Depletion of GGA3
stabilizes BACE and enhances beta-secretase activity. Neuron.54(5):721-37.
13. Kim DY, Carey BW, Wang H, Ingano LA, Binshtok AM, Wertz MH, Pettingell
WH, He P, Lee VM, Woolf CJ, Kovacs DM. (2007). BACE1 regulates
voltage-gated sodium channels and neuronal activity. Nature Cell Biol.
9(7):755-64.
14. Yue X, Lu M, Lancaster T, Cao P, Honda S, Staufenbiel M, Harada N, Zhong Z,
Shen Y, Li R. (2005). Brain estrogen deficiency accelerates Abeta plaque formation
in an Alzheimer's disease animal model. Proc Natl Acad Sci U S A.
102(52):19198-203.
15. Li R., Yang LB, Lindholm K, Yan R, Citron M, Beach T, Sue L, Subbagh M., Cai
H., Wong P, Price D, Shen Y.(2004). Aβ load is correlated with elevated BACE
activity in sporadic Alzheimer patients. Proc Natl Acad Sci U S A.),101(10),
3632-3637.
16. Li R., Lindholm K, Yang LB, Konishi Y, Hampel H, Zhang, D. Shen Y. (2004).
TNF death receptor signaling cascade is required for amyloid-β-protein induced
neuron death. J.Neurosci, 28(7), 1760-1771.
17. Yang LB, Lindholm K, Yan R, Citron M, Xia W, Beach T, Sue L, Wong P, Price D,
Li R, Shen Y. (2003). Elevated beta-secretase expression and enzymatic activity
detected in sporadic Alzheimer disease. Nature.Med, 9(1): 3-4
18. Yang, L-B., Lindholm, K., Konishi, Y., Li, R. and Shen, Y. (2002) Targeting
deletion of TNF receptor subtypes reveals hippocampal neuron survival and death
through distinct signal transduction pathways. J.Neurosci, 22(8), 3025-3032.
19. Yang, L-B., Li, R., Meri, S., Rogers, J. and Shen, Y. (2000) Deficiency of
complement protective protein, CD59 in Alzheimer’s brains may contribute to
neurodegeneration. J.Neurosci, 20(20), 7505-7509.

ABSTRACT
From Bench to Bedside:
BACE1, the Beta-Secretase Enzyme, in Basic Science and Clinical
Investigation
Yong Shen
Robert Haldeman Laboratory of Molecular and Cellular Neurobiology
Banner Sun Health Research Institute
β-Secretase, or β-site amyloid precursor protein cleaving enzyme 1 (BACE1), has been
identified as the rate-limiting enzyme for amyloid-beta (Aβ) peptide production that
occurs primarily in neurons in the brain. Aβis the main component of amyloid plaques
and vascular deposits in Alzheimer's disease (AD) brains, and it is believed to initiate
the deadly amyloid cascade that leads to neurodegeneration and dementia. BACE1 is
the principle beta-secretase, as its knock-out completely prevents Aβ generation.
BACE1 is likely to process a number of different substrates, and consequently, it may
exert several independent physiological functions in neurons. Currently, however, the
physiological functions of BACE1 are not clear. Multiple cellular mechanisms for
BACE1 regulation, including transcriptional regulation, neuronal excitability, receptor
mediation, and identification of new molecules as biomarkers for AD and molecular
targeting for Aβ lowering therapies will be discussed.

INVITED SPEAKER’S INFO
Prof. Craig Montell
PhD, Professor, Director
Departments of Biological Chemistry and Neuroscience
The Johns Hopkins University School of Medicine
855 N. Wolfe St. Baltimore, MD 21205
Tel: (410) 955-1199;
FAX: (410) 287-7672
E-mail: cmontell@jhmi.edu
Education
1978 B.A., Department of Bacteriology, University of California, Berkeley
1983 Ph.D., Department of Microbiology, University of California, Los Angeles
Honorary Degree
2010 March Catholic University, Leuven, Belgium
Appointments
1983-1988 Postdoctoral Fellow, Dept. of Biochemistry,
University of California, Berkeley, CA.
1988-1994 Assistant Professor,Dept. of Biological Chemistry &Neuroscience,
The Johns Hopkins University, School of Medicine, Baltimore, MD.
1994-1998 Associate Professor, Dept. of Biological Chemistry &Neuroscience,
The Johns Hopkins University, School of Medicine, Baltimore, MD.
2000-2006 Director, BCMB Graduate Program
1998- Professor, Dept. of Biological Chemistry &Neuroscience,
The Johns Hopkins University, School of Medicine, Baltimore, MD.
Selected Publications over 100
1. Montell, C., Fisher, E.F., Caruthers, M.H. and Berk, A.J. 1982. Resolving the
functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice
site. Nature 295, 380-384.
2. Montell, C., Fisher, E.F., Caruthers, M.H. and Berk, A.J. 1983. Inhibition of RNA
cleavage but not polyadenylation by a point mutation in mRNA 3' consensus
sequence AAUAAA. Nature 305, 600-605.
3. Montell, C. 2000. PLC fills a GAP in G protein coupled signaling. Nature Cell
Biol. 2, E82-83.
4. Montell, C. 2000. A PDZ protein Ushers in new links. 2000. Nature Genet. 26,
6-7.
5. Montell, C., Courtois, G., Eng, C., and Berk, A.J. 1984. Complete transformation
by adenovirus 2 requires both E1A proteins. Cell 36, 951-961.
6. Montell, C. and Rubin, G. 1988. The Drosophila ninaC locus encodes two

photoreceptor cell specific proteins with domains homologous to protein kinases
and the myosin heavy chain head. Cell 52, 757-782.
7. Bloomquist, B. T., Shortridge, R. D., Schneuwly, S., Perdew, M., Montell, C.,
Steller, H., Rubin, G. and Pak, W. L. 1988. Isolation of a putative phospholipase C
gene of Drosophila, norpA, and its role in phototransduction. Cell 54, 723-733.
8. Xu, X.Z., Li, H.S., Guggino, W.B. and Montell, C. 1997. Coassembly of TRP and
TRPL produces a distinct store-operated conductance. Cell 89, 1155-1164.
9. Montell, C., Birnbaumer, L. and Flockerzi, V. 2002. The TRP channels, a
remarkably functional family. Cell 108, 595-598.
10. Montell, C. 2005. The latest waves in calcium signaling. Cell 122, 157-163.
11. Montell, C. 2007. Dynamic regulation of the INAD signaling scaffold becomes
crystal clear. Cell 131, 19-21.
12. Venkatachalam, K., Long, A.A., Elsaesser, R., Nikolaeva, D., Broadie, K. and
Montell, C. 2008. Motor deficit in a Drosophila model of mucolipidosis type IV
due to defective clearance of apoptotic cells. Cell 135, 838-851.
13. Montell, C. 2010. Preventing a perm with TRPV3. Cell (in press)
14. Montell, C., Jones, K., Hafen, E., and Rubin, G. 1985. Rescue of the Drosophila
phototransduction mutation trp by germline transformation. Science 230,
1040-1043.
15. Porter, J.A., Yu, M., Doberstein, S.K., Pollard, T.S. and Montell, C. 1993.
Dependence of calmodulin localization in the retina on the NINAC unconventional
myosin. Science 262, 1038-1042.
16. Arnon, A., Cook, B., Montell, C., Selinger, Z. and Minke, B. 1997. Calmodulin
regulation of calcium stores in phototransduction of Drosophila. Science 275,
1119-1121.
17. Shen, W., Kwon, Y. and Montell, C. 2010. Rhodopsin required for temperature
discrimination in Drosophila. Science (in revision)
18. Kwon, Y, Shim, H.S., Wang, X. and Montell, C. 2008. Control of thermotactic
behavior via coupling of a TRP channel to a phospholipase C signaling cascade.
Nat. Neurosci. 11, 871-873.
19. Lee, Y., Moon, S.J. and Montell, C. 2009. Multiple gustatory receptors required for
the caffeine response in Drosophila. Proc. Natl. Acad. Sci. U.S.A. 106, 4495-4500.
20. Jiao, Y., Moon, S.J. and Montell, C. 2007. A Drosophila gustatory receptor
required for the responses to sucrose, glucose, and maltose identified by mRNA
tagging. Proc. Natl. Acad. Sci. U.S.A. 104, 14110-14115
21. Liu, Z, Wang, X, Yu, Y, Li, X., Wang, T., Jang, H., Ren, Q., Jiao, Y., Sawa, A.,
Moran, T., Ross, C.A., Montell, C. and Smith, W.W. 2008. A Drosophila model for
LRRK2-linked Parkinsonism. Proc. Natl. Acad. Sci. U.S.A. 105, 2693-2698.
22. Elsaesser, R., Kalra, D., Li, R. and Montell, C. 2010. Light-induced translocation of
Drosophila visual Arrestin2 depends on Rac2. Proc. Natl. Acad. Sci. U.S.A. 107,
4740-4745.
23. Kim, S. H., Lee, Y., Akitake, B., Woodward, O. M., Guggino, W. B. and Montell,
C. 2010. Drosophila TRPA1 channel mediates chemical avoidance in gustatory
receptor neurons. Proc. Natl. Acad. Sci. U.S.A. (in press)

ABSTRACT
TRPA1: a Sentinel for Noxious Chemicals and Temperatures
Craig Montell
Department of Biological Chemistry, The Johns Hopkins University School of Medicine
TRP channels endow us with the ability to sense tastants, mechanical stimuli, a variety
of chemicals, distinct temperatures and in some animals, light and pheromones. In this
seminar, I will begin with the identification of the founding member of the TRP family,
which functions in fly phototransduction, and the realization that TRP channels are
global and evolutionarily conserved mediators of sensory signaling in all animals.
I will describe our recent studies demonstrating that the ability of Drosophila larva to
discriminate between the optimal temperature of 18°C and slightly higher temperatures
depends on the TRPA1 channel, which functions downstream of a
phototransduction-like cascade. We propose that activation of TRPA1 via a signaling
cascade, rather than through direct thermal activation, promotes amplification of small
differences in temperature and facilitates adaptation to temperatures within the
comfortable range.
I will summarize our work demonstrating that insect TRPA1 is a target for insect
repellents. Our studies identifies insect TRPA1 as a potential target for developing
improved repellents to reduce insect-borne diseases such as malaria, which cause
significant morbidity and mortality.

INVITED SPEAKER’S INFO
Prof. Huaxi Xu
PhD, Professor and Acting Director
Neurodegenerative Disease Program
Center for Neuroscience, Stem Cell and Aging Research
Burnham Institute for Medical Research
10901 N. Torrey Pines Road
La Jolla, California 92037.
Tel: 858-795-5246
Fax:858-597-5273
E-mail: xuh@burnham.org
Education
1981-1985 Xiamen University, China. B.S. in Biochemistry.
1987-1988 Visiting Scholar. Division of Rheumatology.
University of Connecticut Health Center, Farmington, CT.
1988-1993 Ph.D. Dept. of Anatomy and Structural Biology.
Albert Einstein College of Medicine, New York.
1993-1994 Post-Doctoral Fellow. Dept.of Developmental and Molecular Biology,
Albert Einstein College of Medicine.
1994-1998 Post-Doctoral Associate, Research Associate.
The Rockefeller University, New York, NY.
Joint Appointment in Dept. of Neurology & Neuroscience,
Cornell University Medical College, New York, NY.
1998-2003 Assistant Professor, Fisher Center for Research on Alzheimer Disease,
The Rockefeller University, New York, NY.
2003-2008 Associate Professor, Neurodegenerative Disease Program
Burnham Institute for Medical Research
2008- Professor and Acting Director,Neurodegenerative Disease Program
Burnham Institute for Medical Research
Selected Publications
1. Xu, H., Sweeney, D., Greengard, P., and Gandy, S. (1996). Metabolism of
Alzheimer ß-Amyloid Precursor Protein: Regulation by Protein Kinase A in Intact
Cells and in a Cell-Free System. Proc. Natl. Acad. Sci. USA. 93:4081-4084.
2. Xu, H., Sweeney, D., Wang, R., Thinakaran, G., Lo, A. C. Y., Sisodia, S.,
Greengard, P., and Gandy, S. (1997) Generation of Alzheimer ß-amyloid
Protein in the trans-Golgi Network in the Apparent Absence of Vesicle Formation.
Proc. Natl. Acad. Sci. USA. 94:3748-3752.
3. Xu, H., Gouras, G. K., Greenfield, J. P., Vincent, B, Naslund, J., Mazzarelli, L,
Jovanovic, J. N, Seeger, M, Relkin, N. R, Liao, F., Checler, F., Buxbaum, J. D,
Chait, B. T., Thinakaran, G., Sisodia, S., Wang, R., Greengard, P., and Gandy, S.
(1998). Estrogen Reduces Neuronal Generation of Alzheimer ß-amyloid Peptides.
Nature.Med 4:447-451.

4. Greenfield, J. P., Tsai, J., Gouras, G. K., Hai, B., Thinakaran, G., Checler, F.,
Sisodia, S. S., Greengard, P., and Xu, H. (1999). Endoplasmic Reticulum and
trans-Golgi Network Generate Distinct Populations of Alzheimer ß-Amyloid
Peptides. Proc. Natl. Acad. Sci. USA 96:742-747.
5. Gouras, G. K.*, Xu, H.*, Gross, R., Greenfield, J. P., Hai, B., Wang, R., and
Greengard, P. (2000) Testosterone Reduces Neuronal Secretion of Alzheimer
ß-amyloid Peptides. Proc. Natl. Acad. Sci. USA 97:1202-1205. (*, equal
contribution).
6. Dou, F., Netzer, W. J., Tanemura, K., Li, F., Hartl, F.U., Takashima, A., Gouras
G.K., Greengard, P., and Xu, H. (2003) Chaperones increase association of tau
protein with microtubules. Proc. Natl. Acad. Sci. USA 100:721-726.
7. Netzer, W.J., Dou, F., Cai, D., Veach, D., Jean, S., Li, Y., Bornmann, W.G.,
Clarkson, B., Xu, H., and Greengard, P. (2003) Gleevec inhibits -amyloid
production but not Notch cleavage. Proc. Natl. Acad. Sci. USA
100(21):12444-12449..
8. Cai, D., Zhong, M., Wang, R., Netner, W.J., Shields, D., Zheng, H., Sisodia S.S.,
Foster, D.A., Gorlick, S.F., Xu, H.*, and Greengard, P*. (2006) Phospholipase D1
corrects impaired APP trafficking and neurite outgrowth in familial Alzheimer’s
disease-linked presenilin-1 mutant neurons. Proc. Natl. Acad. Sci. 103:1936-1940.
9. Li, H.L., Wang, H.H., Liu, S., Deng, Y.Q., Zhang, Y.J., Tian, Q., Wang, X.C.,
Chen, X.Q., Yang, Y., Zhang, J.Y., Wang, Q., Xu, H., Liao, F.F., and Wang, J.Z.
(2007) Phosphorylation of tau antagonizes apoptosis by stabilizing -catenin. Proc.
Natl. Acad. Sci. 104:3591-3596.
10. Zhang, Y-w., Wang, R., Liu, Q., Zhang, H., Liao, F-F., and Xu, H. (2007)
Presenilin/ -secretase-dependent processing of APP regulates EGFR expression.
Proc. Natl. Acad. Sci. 104:10613-10618.
11. Liu, Y., Zhang, Y-w., Wang, X., Zhang, H., You, X., Liao, F-F. and Xu, H. (2009)
Intracellular trafficking of Presenilin 1 is regulated by -amyloid precursor protein
and phospholipase D1. J. Biol. Chem. 284(18):12145-52.
12. Ma, T., Zhao, Y.B., Kwak, Y-D., Thompson, R., Luo, Z., Xu, H., and Liao, F-F.
(2009) Statin's excitoprotection is mediated by sAPP and the subsequent attenuation
of calpain-induced truncation events likely via Rho-ROCK signaling. J. Neurosci.
29:11226-36.
13. Zhang, Y-w., Liu, L., Zhang, X., Li, W-B., Chen, Y., Huang, X., Sun, L., Luo,
W-j., Netzer, W.J., Threadgill, R., Wiegand, G., Wang, R., Cohen, S.N.,
Greengard, P., Liao, F-F., Li, L., and Xu, H. (2009) A Functional Mouse
Retroposed Gene Rps23r1 Inhibits Alzheimer’s -Amyloid Generation and Tau
Phosphorylation. Neuron. 64(3):328-340.
14. Dunys, J., Sevalle, J., Giaimei, E., Pardossi-Piquard, R., Vitek, M.P., Renbaum,
P., Levy-Lahad, E., Zhang, Y-w., Xu, H., Checler, F., and Alves da Costal, C.
(2009) p53-dependent control of PEN-2 promoter transactivation by presenilins. J.
Cell. Sci. 122:4003-4008.
15. Zhang, Y-w., and Xu, H. (2010) Substrate check for gamma-secretase. Nat. Struct.
& Mol. Biol. 17(2):140-141.

ABSTRACT
Novel Genes and Pathways that Regulate Alzheimer’s Disease
Pathogenesis
Huaxi Xu
Neurodegenerative Disease Research Program, Sanford-Burnham Medical Research
Institute,10901 N. Torrey Pines Rd.,La Jolla, CA 92037
Alzheimer's disease (AD) is the most common neurodegenerative disorder, afflicting
millions of aged people in the world. Extraneuronal plaques consisting of β-amyloid
(Aβ) peptides, intracellular neurofibrillary tangles (NFTs) composed of
hyperphosphorylated paired helical filaments of the tau protein, and extensive neuronal
loss are major pathological hallmarks of AD. Multiple lines of evidence suggest that
overproduction/aggregation of the neurotoxic Aβ peptide in the brain is a prime culprit
in AD pathogenesis, causing dramatic neuronal death directly responsible for the
cognitive decline in AD patients. Hyperphosphorylated tau has also been found in many
other neurodegenerative diseases (tauopathies). Hence, knowledge of mechanisms
underlying Aβ generation, tau hyperphosphorylation, and neuronal cell death is crucial
for disease intervention; identification of new genes/proteins involved in these
processes is a major goal of AD research. Our group has recently identified two new
players important in AD: 1) RPS23R1, whose gene originated through retroposition of
the ribosomal protein S23 gene, can reduce Aβ level and tau phosphorylation in cell
cultures and alleviate these AD-like pathologies in the APP/PS1/tau triple transgenic
AD mouse model. Detailed mechanistic studies revealed that RPS23R1 can interact
with adenylate cyclases to increase cAMP synthesis, which upregulates PKA activity,
inhibiting GSK-3 activity which is important in modulating Aβ generation and tau
phosphorylation. 2) Appopstosin is an APP-interacting protein that we identified
through yeast-two-hybrid screens. When overexpressed, appoptosin can induce intrinsic
caspase-dependent apoptosis. Importantly, we find that the level of appoptosin is
upregulated in neurons treated with Aβ, glutamate, or ischemia insults as well as in
AD brains. Furthermore, downregulation of appoptosin can protect neurons against
Aβ- and glutamate-induced neurotoxicity.
Together our studies demonstrate that RPS23R1 is beneficial for inhibiting Aβ
generation and tau phosphorylation, whereas appoptosin-mediated apoptosis plays an
important role in neurotoxicity-induced cell death and neurodegenerative diseases.
These studies will provide new therapeutic possibilities for AD treatment.

INVITED SPEAKER’S INFO
Prof. Jiu-ping Ding
PhD, Professor,
Key laboratory of Molecular Biophysics of the Ministry of
Education, College of Life Science and Technology
Huazhong University of Science and Technology
1037 Luoyulu St.,Wuhan, Hubei, 430074 P.R.China
Tel: 86-27-8779-2153
Fax: 86-27-8779-2024
Email: jpding@mail.hust.edu.cn
Education
1982 M.S (Astrophysics), Nanjing University, Nanjing, China.
1968 B.S. (Electronics), University of Science and Technology of China, Beijing,
China.
Working experiences
1985-1987 Associated Professor, Nanjing Railway Medical College.
1987-1989 Visiting Scholar, SUNY at Buffalo, NY, USA
1989-1992 Research Associated, Dept. of biology,
Washington University, St. Louis, MO, USA
1992-2002 Research Associated, Dept of Anaesthesiology,
Washington University School of Medicine, St. Louis, MO, USA
2002- Professor,
Huazhong University of Science and Technology, Wuhan, China.
Research Experience
1992- Research associate in Dept. of Anesthesiology,
Washington University School of Medicine, St. Louis. MO.
1989-1992 Research associate in Dept. of Biology,
Washington University, St. Louis, MO.
1987-1989 Visiting Scholar in Biophysics,
State University of New York at Buffalo, NY.
Selected Publications
1. Liang Sun, Yu Xiong, Xuhui Zeng, Ying Wu, Na Pan, Christopher J. Lingle, Anlian
Qu and Jiuping Ding. Differential Regulation of Action Potentials by Inactivating
and Noninactivating BK Channels in Rat Adrenal Chromaffin Cells. Biophys. J.,
2009, (In press).
2. Ying Wu, Yu Xiong, Sheng Wang, Hong Yi, Hui Li, Na Pan, Frank T Horrigan
Yingliang Wu and Jiuping Ding. Intersubunit coupling in the Pore of BK channels.
J. Biol. Chem., 2009,284(35): 23353-23363.
3. Zhaohua Guo, Caixia Lv, Hong Yi, Yu Xiong, Yingliang Wu, Wenxin Li, Tao Xu

and Jiuping Ding. A residue at the cytoplasmic entrance of BK-type channels
regulating single-channel opening by its hydrophobicity. Biophys. J., 2008, 4 (9):
3714-3725.
4. Lv, C.X., Chen, M.R., Gan, G.L., Wang, L.F. Xu, T. and Ding, J.P.. Four-turn a
helical segment prevents surface expression of an auxiliary hβ2 subunit of BK-type
channel. J. Biol. Chem., 2008, 283(5): 2709-2715.
5. Geliang Gan, Hong Yi, Maorong Chen, Liang Sun, Wenxin Li, Yingliang Wu and
Jiuping Ding. Structural basis for toxin-resistance of 4 associated
calcium-activated potassium (BK) channels. J. Biol. Chem., 2008, 283:
24177-24184.
6. Maorong Chen, Geliang Gan, Ying Wu, Lu Wang, Yingliang Wu and Jiuping Ding.
Lysine-rich extracellular gate formed by hβ2 subunits reduces the single-channel
conductance of BK channels. PLoS ONE,2008,
7. Shang-bang GAO, Ying WU, Cai-xia LÜ, Zhao-hua GUO, Chen-hong LI, Jiu-ping
DING. Slack and Slick KNa channels are required for the depolarizing
afterpotential of acutely isolated, medium diameter rat dorsal root ganglion neurons.
Acta Pharmacologica Sinica, 2008, 29(8): 899-905.
8. Xiao-Hui Sun, Jiu-Ping Ding, Hui Li, Na Pan, Lu Gan, Xiang-Liang Yang, and
Hui-Bi Xu. Activation of Large-Conductance Calcium-Activated Potassium
Channels by Puerarin: The Underlying Mechanism of Puerarin-Mediated
Vasodilation. J Pharmacol Exp Ther, 2007, 323(1): 391-397.
9. Li, H., Yao, J., Tong, X.T., Wu, Y., Sung, L., Pan, N., Guo, Z.H., Wu, H. M., Xu, T.
and Ding, J.P.. Interaction Sites Between the Slo1 Pore and the N-terminus of the
β2 Subunit, Probed with a Three Residue Sensor. J. Biol. Chem.,2007,282(24):
17720-8.
10. Li, W., Gao, S.B., Lv, C.X., Wu, Y., Guo, Z.H., Ding, J.P. and Xu, T.
Characterization of voltage- and Ca 2+ -activated K + channels in rat dorsal root
ganglion neurons. J. Cell Physiol.,2007,212: 348-57.

ABSTRACT
Structural Feature and Function of hβ-subunits of BK Channel
Jiuping Ding
Key Laboratory of Molecular Biophysics Ministry of Education, College of Life
Science and Technology, Huazhong University of Science and Technology, Wuhan,
Hubei, 430074, China
The functional diversity of large conductance Ca 2+ - and voltage-dependent K + (BK)
channels arises mainly from co-assembly of the pore-forming mSlo α subunits with four
tissue enriched auxiliary β subunits. The structural basis of the interaction between α
subunits with β subunits is not well understood.
Using immunofluorescence and patch clamp techniques, we demonstrated that 1)
four-turn α-helical segment prevents surface expression of the auxiliary hβ2 subunit of
BK-type channel; 2) lysine-rich extracellular rings formed by hβ2 subunits confer the
outward rectification of BK channels; 3) structural basis for toxin resistance of
β4-associated calcium-activated potassium (BK) channels. Those findings provide a
better understanding of the structure and the diverse interactions between α and β
subunits and will improve the design of channel inhibitors.

INVITED SPEAKER’S INFO
Prof. Jianzhi Wang
PhD, Professor, Director,
Professor and Director of Pathophysiology
Pathophysiology Department,
Tongji Medical College,
Huazhong University of Science and Technology
Wuhan, Hubei Province, 430074 P.R.China
Email: wangjz@mails.tjmu.edu.cn
Education and Training
1975-1978 General Medicine,
Tongji (Wuhan) Medical College, Wuhan, China
1985-1988 Post Graduate Training in Biochemistry,
Tongji Medical University, Wuhan, China
1992-1993 Postdoctoral Research in Biochemistry, Siena University, Siena, Italy
1993-1996 Postdoctoral Research in Alzheimer disease,
NYS Institute for Basic Research, SI, NY, USA
Selected Publications
1. Zhu LQ, Zheng HY, Peng CX, Liu D, Li HL, Wang Q and Wang JZ*. Protein
Phosphatase 2A Facilitates Axonogenesis by Dephosphorylating CRMP2. J
Neurosci. 30(10):3839 –48, 2010
2. Zhu LQ, Liu D, Hu J, Cheng J, Wang SH, Wang Q, Wang F, Chen JG, and Wang
JZ*. GSK-3 inhibits presynaptic
vesicle exocytosis by phosphorylating P/Q-type
calcium channel and interrupting SNARE complex formation. J Neurosci.
30(10):3849 –55, 2010
3. Wang JZ*, Liu F. Microtubule-associated protein tau in development, degeneration
and protection of neurons. Prog Neurobiol. 85:148-75, 2008 An invited review
4. Li HL, Wang HH, Liu SJ, Deng YQ, Zhang YJ, Tian Q, Wang XC, Chen XQ,
Yang Y, Zhang JY, Wang Q, Xu H, Liao F and Wang JZ*. Phosphorylation of tau
antagonizes apoptosis by stabilizing -catenin, a mechanism involved in
Alzheimer’s neurodegeneration. Proc Natl Acad Sci U S A. 104(9): 3591-3596,
2007
5. Zhu LQ, Wang SH, Yin YY, Liu D, Zheng HY, Shi HR, Tian Q, Wang XC, Wang
Q, Guo L, Wang JZ*. Activation of glycogen synthase kinase-3 inhibits long term
potentiation with synapse-associated impairments. J Neurosci. 27,12211–20, 2007
6. Wang D, Qian L, Xiong H, Liu J, Neckameyer WS, Oldham S, Xia K, Wang J,
Bodmer R, Zhang Z.Antioxidants protect PINK1-dependent dopaminergic neurons
in Drosophila. Proc Natl Acad Sci U S A. 103:13520-5,2006
7. Liu SJ, Zhang JY, Li HJ, Fang ZY, Wang Q, Deng HM, Gong C, Iqbal K, Wang
JZ*. Tau becomes a more favorable substrate for GSK-3 when it is

prephosphorylated by PKA in rat brain. J Biol Chem. 279(48):50078-88, 2004.
8. Wang JZ, Grundke-Iqbal I, Iqbal K. Glycosylation of microtubule-associated
protein tau: an abnormal posttranslational modification in Alzheimer disease. Nat
Med. 2(8): 870-6, 1996

ABSTRACT
The Synaptic Mechanisms of Glycogen Synathese Kinase-3 in Regulating
Leaning and Memory
Jian-Zhi Wang, Ling-Qiang Zhu, Dan Liu, Juan Hu, Qun Wang
Pathophysiology Department, Key Laboratory of Education Committee on
Neurological Disorders, Tongji Medical College, Huazhong University of Science and
Technology, Wuhan, China
Previous studies have demonstrated that upregulation of glycogen synathese kinase-3
(GSK-3 ) impairs learning
and memory in rats and transgenic mouse models, and
ctivation of GSK-3 inhibits long term potentiation with mechanisms involving a
decreased presynaptic glutamate release. To further verify the mechanisms underlying
the impaired synaptic vesicle exocytosis, we have recently demonstrated that activation
of GSK-3 could inhibit calcium influx through phosphorylating the intracellular loop
connecting domains II and III (LII-III) of P/Q-type Ca 2+ channels, which leads to
decrease of intracellular Ca 2+ influx through the P/Q-type voltage-dependent calcium
channel (VDCC). GSK-3 interferes with the formation of SNARE complex through (i)
weakening the association of synaptobrevin with SNAP25 and syntaxin; (ii) reducing
the interaction between the phosphorylated LII-III and synaptotagmin, SNAP25 and
syntaxin; and (iii) inhibiting dissociation of synaptobrevin from synaptophysin I, all of
which are required for SNARE complex formation and thus for an efficient exocytosis.
These results suggest that GSK-3 regulates
synaptic vesicle cycle by affecting
multiple steps in vesicular exocytosis.

INVITED SPEAKER’S INFO
Prof. XianZhong Xu
PhD, Assistant Professor
Dept of of Molecular & Integrative Physiology
Life Sciences Institute, 6115A
The University of Michigan
Ann Arbor, MI 48109
Tel: (734)615-9311
Email: shawnxu@umich.edu
Education
1987-1991 B.S in Biochemistry , Wuhan University, Wuhan, China ,
1991-1994 M.S in Biochemistry , Wuhan University, Wuhan, China,
1995-2000 MD, Ph.D in Neuroscience& Biochem.
Johns Hopkins University, Baltimore,
2001-2005 Postdoc Neuroscience & Genetics
California Institute of Technology, Pasadena, CA,
Positions and Employment
1991-1993 Guest researcher, National Laboratory of Biomacromolecules,
Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.
2005- Assistant Professor, Dept of Molecular & Integrative Physiology &
Life Sciences Institute, The University of Michigan, Ann Arbor, MI.
Selected Publications
1. Xu, X.Z.S.*, Li, H. S.*, Guggino, W.B., and Montell, C. (1997). Coassembly of
TRP and TRPL produces a distinct store operated conductance. Cell 89, 1155
1164.
2. Xu, X.Z.S., Choudhury, A., Li, X., and Montell, C. (1998). Coordination of an
array of signaling proteins through homo- and heteromeric interactions between
PDZ domains and target proteins. Journal of Cell Biology 142, 545-555
3. Xu, X.Z.S., Wes, P., Li, H. S., Yu, M., Morgan, S., Liu Y., and Montell, C. (1998)
Retinal targets for calmodulin include proteins implicated in synaptic transmission.
J. Biol.Chem 273, 31297-31307.
4. Wes P. D*., Xu, X.Z.S.*, Li, H.-S., Chen, F., Doberstein, S. K., and Montell, C.
(1999) Termination of phototransduction requires binding of the NINAC myosin
III and the PDZ protein INAD. Nature Neuroscience 2, 447-453.
5. Li, H.-S., Xu, X.Z.S., and Montell, C. (1999) Activation of a TRPC3-dependent
cation current through the neurotrophin BDNF. Neuron 24, 261-273.
6. Xu, X.Z.S., Chien, F., Butler, A., Salkoff, L., and Montell, C. (2000) TRP , a
Drosophila TRP-related subunit, forms a regulated cation channel with TRPL.
Neuron 26, 647-657.
7. Xu, X.Z.S., Moebius, F., Gill, D.L., and Montell, C. (2001) Regulation of

melastatin, a TRP-related protein,through interaction with a cytoplasmic isoform.
Proc. Natl. Acad. Sci. U.S.A. 98,10692-10697.
8. Xu, X.Z.S., and Sternberg, P.W. (2003) A C. elegans sperm TRP protein required
for sperm-egg interactions during fertilization. Cell 114, 285-297.
9. Li, W., Feng, Z., Sternberg, P.W., and Xu, X.Z.S. (2006) A C. elegans stretch
receptor neuron revealed by a mechanosensitive TRP channel homologue.
Nature 440, 684-687.
10. Feng, Z.*, Li, W.*, Ward, A., Piggott, B.J., Larkspur, E., Sternberg, P.W., and Xu,
X.Z.S. (2006) A C. elegans model of nicotine-dependent behavior: regulation by
TRP family channels. Cell 127, 621-633
11. Hsu, A.L.*, Feng, Z.*, Hsieh, M.-Y., and Xu, X.Z.S. (2009) Identification by
machine vision of the rate of motor activity decline as a lifespan predictor in C.
elegans. Neurobiology of Aging. 30, 1498-1503. (Epub 2008 Feb 5).
12. Ward, A.*, Liu, J.*, Feng, Z., and Xu, X.Z.S. (2008) Light-sensitive neurons and
channels mediate phototaxis in C. elegans. Nature Neuroscience 11, 916-922.
13. Xiao, R., and Xu, X.Z.S. (2009) Function and regulation of TRP family channels in
C. elegans. Pflugers Arch. 458, 851–860. (review)
14. Ward, A., Walker, V.J., Feng, Z., and Xu, X.Z.S. (2009) Cocaine modulates
locomotion behavior in C. elegans. PLoS ONE 4, e5946. PMCID2691951
15. Liu, J.*, Ward, A.*, Gao, J., Dong, Y., Nishio, N., Inada, H., Kang, L., Yu, Y., Ma.,
D., Xu, T., Mori, I., Xie, Z., and Xu, X.Z.S. (2010) C. elegans phototransduction
requires a G protein-mediated cGMP pathway and a taste receptor homolog.
Nature Neuroscience (in press)

ABSTRACT
Sensory Transduction in C. elegans: What Can't a Worm Sense?
Shawn X. Xu
Life Sciences Institute, University of Michigan, Ann Arbor, USA
The ability to sense and respond to sensory inputs is essential for life. There are five
common senses in mammals: vision, smell, taste, hearing and touch. In addition, we
rely on proprioception, which is often referred to as the sixth sense, to control body
posture, balance and movement. Among the most common sensory stimuli are
chemicals (smell and taste), mechanical forces (touch, hearing and proprioception) and
light (vision). We are particularly interested in understanding how neurons detect and
transduce mechanical and light stimuli, and how gene networks (e.g. receptors, ion
channels and signaling molecules) regulate these processes. Currently, we focus on
mechanotransduction and phototransduction. We have reported that proprioception is
present in C. elegans. We have also made the surprising discovery that C. elegans, an
organism that is generally believed to be blind (light-insensitive), in fact possesses a
simple visual system, senses light and engages in phototaxis behavior. Our data reveal
conservations in mechanotransduction and phototransduction between worms and
mammals. These studies establish C. elegans as a powerful genetic model for studying
the mechanisms of mechanotransduction and phototransduction.

INVITED SPEAKER’S INFO
Prof. Jean-Louis Besserau
Ph.D, M.D, Profesosor
Genetique et Neurobiologie de C. elegans
INSERM U. 789
Ecole Normale Superieure
46, rue d'Ulm,75 005 Paris,France
Lab phone: 33 (0)1 44 32 23 05
Fax: 33 (0)1 44 32 36 54
Email: jlbesse@wotan.ens.fr
Education
1994 M.D., Specialization in Neurology.
1995 Ph.D,Molecular and cellular pharmacology,
University of Paris VI, France.
1995 Researcher position at the INSERM,
Laboratory of Pr Changeux- Pasteur Institute, Paris.
1997 Post-doctoral position in Erik Jorgensen laboratory,
University of Utah, USA.
2000 Junior group leader, Ecole Normale Supérieure-Paris.
2005 INSERM Research Director at the Ecole Normale Supérieure-Paris.
Selected Publications
1. Gendrel M et al.: 2009. A secreted complement-control-related protein ensures
acetylcholine receptor clustering. Nature 461: 992-996.
2. Jospin M et al.: 2009. A Neuronal Acetylcholine Receptor Regulates the Balance of
Muscle Excitation and Inhibition in Caenorhabditis elegans. PLoS Biol 7:
e1000265.
3. Robert VJ and Bessereau JL: 2009. Manipulating the Caenorhabditis elegans
genome using mariner transposons. Genetica.
4. Boulin T et al.: 2008. Eight genes are required for functional reconstitution of the
Caenorhabditis elegans levamisole-sensitive acetylcholine receptor. Proc Natl
Acad Sci U S A 105: 18590-18595.
5. Nilsson L et al.: 2008. Caenorhabditis elegans num-1 Negatively Regulates
Endocytic Recycling. Genetics 179: 375-387.
6. Robert V et al.: 2008. Gene conversion and end-joining repair double-strand breaks
in the C. elegans germline. Genetics 180: 673-679.
7. Ruaud AF et al.: 2008. The C. elegans P4-ATPase TAT-1 regulates lysosome
biogenesis and endocytosis. Traffic 10: 88-100.
8. Eimer S et al.: 2007. Regulation of nicotinic receptor trafficking by the
transmembrane Golgi protein UNC-50. EMBO J 26: 4313-4323.
9. Ruaud AF and Bessereau JL: 2007. The P-type ATPase CATP-1 is a novel

egulator of C. elegans developmental timing that acts independently of its
predicted pump function. Development 134: 867-879.
10. Robert V and Bessereau JL: 2006. Targeted engineering of the Caenorhabditis
elegans genome following Mos1-triggered chromosomal breaks. EMBO J 26:
170-183.
11. Ruaud AF and Bessereau JL: 2006. Activation of nicotinic receptors uncouples a
developmental timer from the molting timer in C. elegans. Development 133:
2211-2222.
12. Weimer RM et al.: 2006. UNC-13 and UNC-10/rim localize synaptic vesicles to
specific membrane domains. J Neurosci 26: 8040-8047.
13. Williams DC et al.: 2005. Characterization of Mos1-mediated mutagenesis in
Caenorhabditis elegans: a method for the rapid identification of mutated genes.
Genetics 169: 1779-1785.
14. Yeh E et al.: 2005. Identification of genes involved in synaptogenesis using a
fluorescent active zone marker in Caenorhabditis elegans. J Neurosci 25:
3833-3841.
15. Gally C et al.: 2004. A transmembrane protein required for acetylcholine receptor
clustering in Caenorhabditis elegans. Nature 431: 578-582.
16. Rostaing P et al.: 2004. Preservation of immunoreactivity and fine structure of adult
C. elegans tissues using high-pressure freezing. Journal of Histochemistry &
Cytochemistry 52: 1-12.
17. Bosher JM et al.: 2003. The Caenorhabditis elegans vab-10 spectraplakin isoforms
protect the epidermis against internal and external forces. J Cell Biol 161: 757-768.
18. Gally C and Bessereau JL: 2003. GABA is dispensable for the formation of
juncational GABA receptor clusters in Caenorhabditis elegans. J.Neurosci 23:
2591-2599.
19. Gally C and Bessereau JL: 2003. C. elegans: of neurons and genes. M S-Medecine
Sciences 19: 725-734.
20. Bessereau J-L et al.: 2001. Mobilization of a Drosophila transposon in the
Caenorhabditis elegans germ line. Nature 413: 70-74.

ABSTRACT
Spatial Control of Synaptic Transmission at a Cholinergic Synapse in
C. elegans
Jean-Louis BESSEREAU
Ecole Normale Superieure, INSERM U789, Paris, France
Chemical synapses are specialized junctions that ensure precise and efficient signal
transduction between excitable cells. Specifically, synaptic vesicles fuse with the
pre-synaptic membrane at the active zone, in register with a specialized domain of the
post-synaptic membrane containing a high density of neurotransmitter receptors.
To analyze the subcellular organization of synapses at high resolution in a near to
living-state, we physically immobilized freely moving nematodes C. elegans by instant
freezing under high-pressure. After freeze-substitution and resin embedding, we used
electron microscopy (EM) tomography to improve 3D resolution as compared to
classical transmission-EM. We observed that at cholinergic neuromuscular junctions
synaptic vesicles were interconnected by filaments similar to the cytomatrix of
vertebrates. Furthermore, vesicles docked at the plasma membrane were retained at the
active zone by physical contacts with the presynaptic dense projection. In unc-10/RIM
and syd-2/liprin mutants, docked vesicles mislocalized, which provided structural basis
for the functional defects observed in these mutants.
On the post-synaptic membrane, ionotropic acetylcholine receptors (AChRs) are
clustered at high density in front of ACh release sites. We previously demonstrated that
clustering depend on the muscle-secreted protein LEV-9 and the transmembrane protein
LEV-10 that assemble in an extracellular scaffold with AChRs. In a forward genetic
screen, we identified a gene coding for a small secreted protein containing only one
immunoglobulin domain. This protein localizes at cholinergic neuromuscular junctions
and is necessary to stabilize the AChR/LEV-9/LEV-10 complex at the synapse.

INVITED SPEAKER’S INFO
Prof. Jian-guo Chen
M.D, Professor
Dept. of Pharmacology, Tongji Medical College,
Huazhong University of Science Technology,
#13 Hangkong Rd, Wuhan, 430030, China
Phone: 0086-27-83692636
Fax: 0086-27-83692608
E-mail: chenj@mails.tjmu.edu.cn
Education and Academic Appointments
2002- Professor, Chairman, Dept. of Pharmacology, Tongji Medical College,
Huazhong University of Science & Technology
1998-2001 Postdoctoral Associate, Dr. T. Hoshi’s laboratory
The University of Iowa College of Medicine,USA.
1996-1998 Visiting scholar, Dr. J. Sandkuhler’s laboratory, II.
The Medical College, University of Heidelberg ,Germany.
1995-1996 Visiting scholar, Prof. Dr. J. W. Deitmer’s laboratory,
The University of Kaiserslautern ,Germany.
1990-1995 Lecturer, Dept. of Pharmacology,
Tongji Medical University ,China.
1987-1990 MD, Prof. ZW Li’s laboratory
The Center of Experimental Medical Researches,
Tongji Medical University ,China.
1980-1985 Bachelor of Medcine, Xianning Medical College ,China.
Selected Publications
1. Huang C, Hu ZL, Wu WN, Yu DF, Xiong QJ, Song JR, Shu Q, Fu H, Wang F and
Chen JG. Existence and distinction of acid-evoked currents in rat astrocytes. Glia
(2010, accepted, in press)
2. Liu C, Wu JL, Xu K, Cai F, Gu J, Ma LQ and Chen JG. Neuroprotection by
baicalein in ischemic brain injury involves PTEN/AKT pathway. J
Neurochemistry, (2010, accepted, in press)
3. Yang MJ, Wang F, Wang JH, Wu WN, Hu ZL, Cheng J, Yu DF, Long LH, Fu H, Xie
N, Chen JG*. PI3-k integrates the effects of insulin and leptin on large-conductance
Ca 2+ -activated K + channels in neuropeptide Y neurons of the hypothalamic arcuate
nucleus. Am J Physiol Endocrinol Metab. 2010, 298(2)E: 193-201.
4. Wang T, Gu J, Wu PF, Wang F, Xiong Z, Yang YJ, Wu WN, Dong LD and Chen JG.
Protection by Tetrahydroxystilbene glucoside against cerebral ischemia:
Involvement of JNK, SIRT1, NF-κB pathways and inhibition of intracellular

ROS/RNS generation. Free Radical Biology & Medicine,2009 Aug
1;47(3):229-40.
5. Mao LM, Wang W, Chu XP, Zhang GC, Liu XY, Yang YJ, Haines M, Papasian CJ,
Fibuch EE, Buch S, Chen JG, and Wang JQ. Stability of NMDA receptors at the
plasma membrane controls synaptic and behavioral adaptations to amphetamine,
Nature Neuroscience, 2009 May;12(5):602-10.
6. Liu J, Wang W, Wang F, Cai F, Hu ZL, Yang YJ, Chen J, Chen JG.
Phosphatidylinositol-linked novel D(1) dopamine receptor facilitates long-term
depression in rat hippocampal CA1 synapses. Neuropharmacology. 2009 Aug;
57(2):164-71.
7. Cai F, Wang F, Lin FK, Liu C, Ma LQ, Liu J, Wu WN, Wang W, Wang JH, Chen JG.
Redox modulation of long-term potentiation in the hippocampus via regulation of
the glycogen synthase kinase-3beta pathway. Free Radical Biology & Medicine,
2008 Oct 1;45(7):964-70.
8. Ma LQ, Liu C, Wang F, Xie N, Gu J, Fu H, Wang JH, Cai F, Liu J and Chen JG,
Activation of phosphatidylinositol-linked novel D1 dopamine receptors inhibits
high voltage activated Ca 2+ currents in primary cultured striatal neurons. Journal of
Neurophysiology, 2009 May;101(5):2230-8.
9. Zhang WT, Jiang FC, Wu PF, Ruan JL, Zhang LN, Fang W, Chen XL, Wang Y, Cao
BS, Chen GY, Zhu YJ, Gu J and Chen JG*. Design, synthesis, and cytoprotective
effect of 2-Aminothiazole Analogs as potent poly (ADP-ribose) polymerase-1
inhibitors. Journal of Medicinal Chemistry, 2009 Feb 12;52(3):718-25.
10. Cai F, Wang F, Lin FK, Liu C, Ma LQ, Liu J, Wu WN, Wang W, Wang JH, Chen JG.
Redox modulation of long-term potentiation in the hippocampus via regulation of
the glycogen synthase kinase-3beta pathway. Free Radical Biology & Medicine,
2008 Oct 1;45(7):964-70.
11. Wang JH, Wang F, Yang MJ, Yu DF, Wu WN, Liu J, Ma LQ, Cai F, Chen JG. Leptin
regulated calcium channels of neuropeptide Y and proopiomelanocortin neurons by
activation of different signal pathways. Neuroscience. 2008 Sep 22;156(1):89-98.
12. Lin F, Xin Y, Wang J, Ma L, Liu J, Liu C, Long L, Wang F, Jin Y, Zhou J, Chen J
(2007). Puerarin facilitates Ca 2+ -induced Ca 2+ release triggered by
KCl-depolarization in primary cultured rat hippocampal neurons. European
Journal of Pharmacology, 570:43-49
13. Lin FK, Xin Y, Gao DM, Xiong Z and Chen JG *(2007). Effects of electrical
stimulation of the parafascicular nucleus neuronal activities of the subthalamic
nucleus and the ventromedial nucleus in rats. Acta Physiologica Sinica
59(1):79-85.
14. Moore SA, Saito F, Chen J, Michele DE, Henry MD, Messing A, Cohn RD,
Ross-Barta SE, Westra S, Willamson R, Hoshi T and Campbell KP. 2002.
Deletion of brain dystroglycan recapitulates aspects of congenital muscular
dystrophy. Nature, Vol. 418: 422-425.

ABSTRACT
The Function of Phosphatidylinositol-Linked Novel D1 Dopamine
Receptor in Central Nervous System
Jian-Guo Chen, Fang Wang, Jue Liu and Li-Qun Ma
Department of Pharamcology and the Key Laboratory of Neurological Disease of
Ministry of Education of China , Tongji Medical College, Huazhong University of
Science and Technology, Wuhan, 430030, China
The physiological actions of dopamine (DA) are mediated via five distinct subtypes
(D1-D5) of G protein-coupled receptors. Stimulation (D1, D5 subtypes) or inhibition
(D2-D4 subtypes) of adenylyl cyclase through Gs/olf or Gi/o proteins by DA receptors
is the traditional signaling pathway. In addition to the Gs/olf-coupled classical D1 DA
receptor that increases the formation of cAMP, a D1-like DA receptor that couples to
Gq protein, stimulates phospholipase Cb (PLCb) and results in hydrolysis of
phosphoinositide has been described and named as PI-linked DA receptor. SKF83959, a
recently identified selective agonist for the PI-linked DA receptor, provided a powerful
tool for exploring the function of this novel signal pathway in brain. However, the
physiological and pathological roles of SKF83959 are unclear. In the recent work from
our laboratory, it has been shown that activation of D1 receptor by SKF83959 mediates
a dose-dependent mobilization of [Ca 2+ ]i via the PLC signaling pathway in cultured rat
prefrontal cortical neurons and astrocytes. It was also inhibited HVA Ca2+ currents in
cultured rat striatal neurons by PLC-IP3-Ca2+-calcineurin signal pathway. Furthermore,
SKF83959 facilitated a postsynaptically PLC-dependent form of LTD at the Schaffer
collater-CA1 synapses. In the pathological condition, SKF83959 mediates more
effective functions of anti-Parkinsonian symptom with less dyskinesia. Our results
provide further understanding for the role of PI-coupled dopamine system that may
mediate important physiological or pathological challenges in the brain.

INVITED SPEAKER’S INFO
Prof. Xi He
PhD, Professor of Neurology, Harvard Medical School
Children's Hospital Boston
300 Longwood Avenue,Boston, MA, 02115
Phone: 617-919-2257
Fax: 617-730-1953
Email: xi.he@childrens.harvard.edu
Education
1982-1985 Master’sdegree, Department of Biomedical Engineering,
Huazhong University of Science and Technology, China.
1985-1986 Research Associate, Department of Biomedical Engineering,
Huazhong University of Science and Technology, China.
1986-1992 Graduate Studies with Dr. Michael G. Rosenfeld, HHMI,
School of Medicine, University of California at San Diego ,La Jolla.
1992-1994 Postdoctoral Fellow with Dr. Luis F. Parada, NCI/NIH ,Frederick.
1994-1996 Postdoctoral Fellow with Dr. Harold E.Varmus, NCI/NIH ,Bethesda.
Professinal Experience
1997 Assistant Professor, Neuroscience, Children’s Hospital,
Harvard Medical School (Boston, MA)
2002 Associate Professor, Neuroscience, Children’s Hospital,
Harvard Medical School (Boston, MA)
2007 Professor, Neurology/Neuroscience, Children’s Hospital,
Harvard Medical School (Boston, MA)
Selected Publications
1. He X, Treacy M, Simmons D, Ingraham H, Swanson L, Rosenfeld Mg. Expression
of a Large Family of POU-domain Regulatory Genes in Mammalian Brain
Development. Nature 1989; 340:35-42.
2. He X, Saint-Jeannet JP, Woodgett JR, Varmus HE, Dawid IB. Glycogen synthase
kinase-3 and dorsoventral patterning in Xenopus embryos. Nature 1995;
374:617-622.
3. He X, Saint-Jeannet JP, Nathas J, Wang Y, Dawid IB, Varmus HE. A member of
the frizzled protein family mediating axis induction by Wnt-5A. Science 1997;
275:1652-1654.
4. Tamai K, Semenov M, Kato Y, Spokony R, Liu C, Katsuyama Y, Hess F,
Saint-Jeannet J-P, He X. LDL receptor-related proteins in Wnt signal transduction.
Nature 2000; 407: 530-535.
5. Semenov M, Tamai K, Brott B, Kuhl M, Sokol S, He X. Head inducer Dickkopf-1

is a ligand for Wnt coreceptor LRP6. Current Biology 2001; 11: 951-961.
6. Habas R, Kato Y, He X. Wnt/Frizzled activation of Rho regulates vertebrate
gastrulation and requires a novel Formin homology protein Daam1. Cell 2001; 107,
843-854.
7. Liu C, Li Y, Semenov M, Baeg G-H, Han C, Tan Y, Zhang Z, Lin X, He X.
Control of beta-catenin phosphorylation-degradation by a dual-kinase mechanism.
Cell 2002; 108, 837-847.
8. Kato Y, Habas R, Katsuyama Y, Naar A, He X. A component of the ARC/Mediator
complex required for TGF-beta/Nodal signaling. Nature 2002; 418: 641-646.
9. Capelluto DGS, Kutateladze TG, Habas R, Finkielstein CV, He X, Overduin M.
DIX domain targets Dishevelled to actin stress fibers and vesicular membranes.
Nature 2002; 419, 726-729.
10. Habas, R., Dawid, I., and He, X. Co-activation of Rac and Rho by Wnt/Frizzled
signaling is required for vertebrate gastrulation. Genes Dev. 2003; 17, 295-309.
11. He, X. Wnt signaling went derailed again: a new track via the LIN-18 receptor?
Cell 2004, 118, 668-670.
12. He, X. Antagonizing Wnt and FGF receptors: an enemy from within (the ER). Cell
2005 120, 156-158.
13. Zeng X, Tamai K, Doble B, Li S, Huang H, Habas R, Okamura H, Woodgett J and
He X. A dual-kinase mechanism for Wnt coreceptor phosphorylation and activation.
Nature, 2005, 438, 873-877.
14. He X and Axelrod J. A WNTer wonderland in Snowbird. Development, 2006, 133,
2597-2603.
15. He X. Unwinding a path to nuclear beta-catenin. Cell, 2006, 127, 40-42.
16. MacDonald B, Semenov M and He X. Wnt/beta-catenin signaling (SnapShot). Cell
2007, 131, 1204-1205.
17. Semenov M, Habas R, MacDonald B, and He X. Non-canonical Wnt signaling
(SnapShot). Cell 2007, 131, 1378-1379.
18. Zeng X, Huang H, Tamai K, Zhang X, Harada, Y, Yokota, C, Almeida, KL, Wang
J, Doble B, Woodgett J, Wynshaw-Boris A, Hsieh J-C, He X. Initiation of Wnt
signaling: Control of Wnt coreceptor LRP6 phosphorylation/activation via
Frizzled/Dishevelled/Axin Functions. Development .
19. Choi YJ, Di Nardo A, Kramvis I, Meikle L, Kwiatkowski DJ, Sahin M, He X.
Tuberous Sclerosis Complex proteins control axon formation. Genes & Dev. 2008,
22, 2485-2495.
20. Wu G, Huang H, Abreu JG, He X. Inhibition of GSK3 phosphorylation of b-catenin
via phosphorylated PPPSPXS motis of Wnt receptor LRP6. PLoS One 2009, 4,
e4926.

ABSTRACT
Mechanisms of Wnt Signaling and Regulation
Xi He, Xinjun Zhang, Jose Garcia Abreu, Chika Yokota, Bryan MacDonald, He Huang
The F. M Kirby Center for Neurobiology, Children’s Hospital Boston, CLS-12064,
Harvard Medical School, Boston, MA 02115, USA.
Canonical Wnt/β-catenin signaling is initiated by the action of the Frizzled (Fz) receptor
and its coreceptor LDL receptor-related-protein 6 (LRP6). Wnt signaling induces LRP6
phosphorylation at conserved PPPSPxS motifs, which serve as docking sites for the
scaffolding protein Axin, thereby allowing the Wnt receptor complex to inhibit
b-catenin phosphorylation and degradation. LRP6 phosphorylation is mediated via the
action of glycogen synthase kinase 3 (GSK3) and casein kinase 1 (CK1), and requires
the function of the Fz receptor and its downstream partner Dishevelled (Dvl).
I’ll discuss our study on the regulation of LRP6 phosphorylation, and the role of
LRP6-Axin interaction in the initiation and amplification of Wnt signaling at the plasma
membrane. I’ll also discuss some biochemical experiments aimed to understand the
mechanism by which phosphorylated LRP6 leads to inhibition of b-catenin
phosphorylation. Finally I’ll describe a novel transmembrane protein, Tiki1, that
appears to regulate the Wnt morphogen via an unexpected mechanism during vertebrate
anterior patterning.

INVITED SPEAKER’S INFO
Prof. John Wemmie
MD, PhD, Assistant Professor
University of Iowa, Roy J and Lucille A Carver
College of Medicine, Department of Psychiatry
500 EMRB, Iowa City IA 52242
Tel: 319 335 6540
Fax: 319 335 7623
Email: john-wemmie@uiowa.edu
Education
1989 BA, Central College, , Chemistry, Pella, IA
1988 Trinity College, Foreign Study
Carmarthen, Wales, UK Program
1992 St. Petersburg State University, Intensive Russian,
St. Petersburg, Russia, Language
1996 PhD, University of Iowa Department of Physiology and Biophysics, Iowa City,
1996 M.D, University of Iowa College of Medicine, Iowa City,
2000 Residency Psychiatry,University of Iowa Hospitals and Clinics
Selected Publications
1. Wemmie J, Chen J, Askwith C, Hruska-Hageman A, Price M, Nolan B, Yoder P,
Lamani E, Hoshi T, Freeman J, Welsh M: The Acid-Activated Ion Channel ASIC
Contributes to Synaptic Plasticity, Learning, and Memory. Neuron (34): 463-477,
2002.
2. Wemmie J, Askwith C, Lamani E, Cassell M, Freeman J, Welsh M: ASIC1 is
Localized in Brain Regions with High Synaptic Density and Contributes to Fear
Conditioning. J Neurosci 23:5496-5502, 2003.
3. Leonard AS, Yermolaieva O, Hruska-Hageman A, Askwith CC, Price MP,
Wemmie JA, Welsh MJ: cAMP-dependent protein kinase phosphorylation of the
acid-sensing ion channel-1 regulates its binding to the protein interacting with
C-kinase-1. Proc Natl Acad Sci U S A. Feb 18;100 (4):2029-34, 2003.
4. Xiong Z, Zhu X, Chu X, Minami M, Hey J, Wemmie JA, Price MP, Welsh MJ,
Simon RP, Neuroprotection in ischemia: blocking calcium-permeable acid-sensing
ion channels, Cell 118(6):687-698, 2004.
5. Wemmie JA, Coryell MW, Askwith CC, Lamani E, Leonard AS, Sigmund CD,
Welsh MJ: Overexpression of acid-sensing ion channel 1a in transgenic mice
increases acquired fear-related behavior. Proc Natl Acad Sci USA. 2004 Mar
9;101(10):3621-6. Epub 2004 Feb 26.
6. Chu XP, Wemmie JA, Wang WZ, Zhu XM, Saugstad JA, Price MP, Simon RP,
Xiong ZG: Subunit-dependent high-affinity zinc inhibition of acid-sensing ion
channels. J Neurosci 2004 Oct 6;24(40):8678-89.

7. Zha X-M, Wemmie JA, Welsh MJ (2006) ASIC1a is a postsynaptic proton receptor
that influences the density of dendritic spines. Proc Natl Acad Sci USA
103:16556-16561.
8. Coryell M, Espie-Ziemann A, Westmoreland P, Haenfler J, Kurjakovic Z, Zha X,
Price M, Schnizler M, Wemmie J. Targeting ASIC1a reduces innate fear and
alters neural activity in the fear circuit. Biological Psychiatry. 2007 Jul 27;
9. Friese MA, Craner MJ, Etzensperger R, Vergo S, Wemmie JA, Welsh MJ, Vincent
A, Fugger L. Acid-sensing ion channel-1 contributes to axonal degeneration in
autoimmune inflammation of the central nervous system. Nat Med. 2007
Dec;13(12):1483-9. Epub 2007 Nov 11.
10. Ziemann AE, Schnizler MK, Albert GW, Severson MA, Howard III MA, Welsh
MJ, Wemmie JA. Seizure termination by acidosis depends on ASIC1a. Nat
Neurosci. 2008 Jul;11(7):816-22. Epub 2008 Jun 8.
11. Coryell M, Wunsch A, Haenfler J, Allen J, McBride J, Davidson B, Wemmie J,
Restoring Acid-Sensing Ion Channel-1a in the Amygdala of Knockout Mice
Rescues Fear Memory but not Unconditioned Fear Responses. J Neurosci. 2008
Dec 17;28(51):13738-41.
12. Oxidant regulated inter-subunit disulfide bond formation between ASIC1a subunits.
Zha XM, Wang R, Collier DM, Snyder PM, Wemmie JA, Welsh MJ. Proc Natl
Acad Sci U S A. 2009 Mar 3;106(9):3573-8.
13. Acid-sensing ion channel-1a in the amygdala, a novel therapeutic target in
depression-related behavior. Coryell MW, Wunsch AM, Haenfler JM, Allen JE,
Schnizler M, Ziemann AE, Cook MN, Dunning JP, Price MP, Rainier JD, Liu Z,
Light AR, Langbehn DR, Wemmie JA. J Neurosci. 2009 Apr 29;29(17):5381-8
14. Ziemann AE, Allen JE, Dahdaleh NS, Drebot II, Coryell MW, Wunsch AM, Lynch
CM, Faraci FM, Howard III MA, Welsh MJ, Wemmie JA. The Amygdala Is a
Chemosensor that Detects Hypercarbia and Acidosis to Elicit Fear Behavior. Cell,
2009 Volume 139, Issue 5, 1012-1021, 25 November 2009
15. Wemmie JA, Price MP, Welsh MJ (2006) Acid-sensing ion channels: advances,
questions and therapeutic opportunities. Trends Neurosci 29:578-586.
16. Kathleen A. Sluka, Olivia C. Winter, John A. Wemmie. Acid-Sensing Ion
Channels: A New Target for Pain and CNS Diseases. Curr Opin Drug Discov
Devel. 2009 Sep;12(5):693-704.
17. J. Render1, K. Howe, A. Wunsch, S. Guionaud, P. Cox, J. Wemmie Histologic
Examination of the Eye of Acid-Sensing Ion Channel 1a Knockout Mice, Int J
Physiol Pathophysiol Pharmacol 2010;2(1):69-72

ABSTRACT
Acid Sensing Ion Channels: A Novel Therapeutic Target for Anxiety and
Depression?
John Wemmie
University of Iowa Carver College of Medicine, USA
Acid sensing ion channels (ASICs) are members of the degenerin/epithelial sodium
channel family that are activated by low extracellular pH. Several ASIC subunits are
expressed in the brain including ASIC1a, 2a, and 2b. of these, our work suggests that
ASIC1a is critical for acid-evoked currents in brain neurons. Disrupting ASIC1a in
mice eliminated currents in CNS neurons evoked by pH as low as 5.0. Subcellular
localization suggests ASIC1a is distributed to the neuron cell body, dendrites, and
dendritic spines, where it confers pH sensitivity, increases intracellular Ca 2+ , and
promotes synaptic plasticity. ASIC1a protein was found to be particularly abundant in
brain structures that underlie fear, anxiety, and depression-related behaviors, including
the basolateral amygdala. Consistent with these observations, loss of ASIC1a
disrupted fear-related learning and memory. Loss of ASIC1a also reduced
unconditioned fear, anxiety, and depression-related behaviors in mice. Supporting a
role for ASIC1a as a brain pH sensor, we found that ASIC1a in the amygdala mediates
fear and anxiety responses to CO2 inhalation and brain acidosis. This observation that
suggests that ASICs may underlie the well-described ability of CO2 to triggers panic
attacks in panic disorder patients. Together, these findings raise the possibility that
ASIC1a might provide a novel and effective therapeutic target for anxiety disorders and
depression.

INVITED SPEAKER’S INFO
Prof. Tao Xu
PhD, Professor, Director
Institute of Biophysics,
Chinese Academy of Sciences
Beijing 100101, P.R. China
Phone: +86-10-64888469
Fax: +86-10-64871293
E-mail: xutao@ibp.ac.cn
Education
1992-1996 Ph.D. (Biophysics), Institute of Biophysics & Biochemistry,
Huazhong University of Science and Technology, China.
1988-1992 B.S. (Engineering),
Huazhong University of Science and Technology, China.
Reaearch Experience
2007- Professor, Director, Institute of Biophysics,
Chinese Academy of Sciences, Beijing, China
2003-2007 Professor, Deputy Director, Institute of Biophysics,
Chinese Academy of Sciences, Beijing, China
2000-2003 Professor, Institute of Biophysics and Biochemistry,
Huazhong University of Science and Technology, China.
1999-2000 Senior Fellow, Department of Physiology and Biophysics,
University of Washington, USA.
1996-1999 Postdoctoral Fellowship,
Max-Planck-Institute for Biophysical Chemistry, Germany.
Selected Publications
1. Wei Ji, Pingyong Xu, Zhengzheng Li, Jingze Lu, Lin Liu, Yi Zhan, Yu Chen, Bertil
Hille, Tao Xu*, And Liangyi Chen*. Functional Stoichiometry of The Unitary
Calcium-Release-Activated Calcium Channel. Proc. Natl. Acad. Sci. (USA).
105(36): 13668–13673. 2008.
2. Zixuan He, Junmei Fan, Lijun Kang, Jingze Lu, Yanhong Xue, Pingyong Xu, Tao
Xu* And Liangyi Chen*. Ca 2+ Triggers A Novel Clathrin-Independent But
Actin-Dependent Fast Endocytosis In Pancreatic Beta Cells. Traffic. 9: 910–923.
2008.
3. Li Jiang, Junmei Fan, Li Bai, Yan Wang, Yu Chen, Lu Yang, Liangyi Chen, And
Tao Xu*. Direct Quantification of Fusion Rate Reveals A Distal Role For As160 In
Insulin-Stimulated Fusion of Glut4 Storage Vesicles. J. Biol. Chem. 283(13):
8508–8516. 2008.
4. Guo Z, Lv C, Yi H, Xiong Y, Wu Y, Li W, Xu T*, Ding J*. A Residue At The

Cytoplasmic Entrance of Bk-Type Channels Regulating Single-Channel Opening
By Its Hydrophobicity. Biophys J. 94(9): 3714-25. 2008.
5. Li Bai, Yan Wang, Junmei Fan, Yu Chen, Wei Ji, Anlian Qu, Pingyong Xu*, David
E. James, And Tao Xu*. Dissecting Multiple Steps of Glut4 Trafficking And
Identifying The Sites of Insulin Action. Cell Metabolism. 5: 47-57. 2007.
6. Ke-Ming Zhou, Yong-Ming Dong, Qian Ge, Dan Zhu, Wei Zhou, Xian-Guang Lin,
Tao Liang, Zheng-Xing Wu*, Tao Xu*. Pka Activation Bypasses The Requirement
For Unc-31 In The Docking of Dense Core Vesicles From C.Elegans Neurons.
Neuron. 56: 657-669. 2007.
7. Zhengzheng Li, Jingze Lu, Pingyong Xu, Xiangyang Xie, Liangyi Chen*, And Tao
Xu*. Mapping The Interacting Domains of Stim1 And Orai1 In
Ca 2+ Release-Activated Ca 2+ Channel Activation. J. Biol. Chem. 282(40):
29448–29456. 2007.
8. Wei Li, Shang-Bang Gao, Cai-Xia Lv, Ying Wu, Zhao-Hua Guo, Jiu-Ping Ding*,
And Tao Xu*. Characterization of Voltage- And Ca 2+ -Activated K + Channels In
Rat Dorsal Root Ganglion Neurons. Journal of Cellular Physiology. 212(2):
348-357. 2007.
9. Lijun Kang, Zixuan He, Pingyong Xu, Junmei Fan, Andrea Bet, Nils Brose, And
Tao Xu*. Munc13-1 Is Required For The Sustained Release of Insulin From
Pancreatic Beta Cells. Cell Metabolism. 3: 463-468. 2006
10. Li Bai, Dan Zhu, Keming Zhou, Wei Zhou, Dongdong Li, Yan Wang, Rongying
Zhang And Tao Xu*. Differential Properties of Gtp- And Ca 2+ -Stimulated
Exocytosis From Large Dense Core Vesicles. Traffic. 7: 416-428. 2006.
11. Xie L, Zhang M, Zhou W, Wu Zx, Ding Jp, Chen Ly*, And Xu T*. Extracellular
Atp Stimulates Cells.�Exocytosis Via Localized Ca 2+ Release From Acidic Stores In
Rat Pancreatic. Traffic. 7: 429-439. 2006.
12. Dongfang Liu, Liang Xu, Fan Yang, Dongdong Li, Feili Gong, And Tao Xu*.
Rapid Biogenesis And Sensitization of Secretory Lysosomes In NK Cells Mediated
By Target-Cell Recognition. Proc Natl Acad Sci U S A. 102(1): 123-127. 2005.
13. Jie Liu, Qunfang Wan, Xianguang Lin, Hongliang Zhu, Kirill Volynski, Yuri
Ushkaryov And Tao Xu*. Α-Latrotoxin Modulates The Secretory Machinery Via
Receptor-Mediated Activation of Protein Kinase C. Traffic. 6: 756- 765. 2005.
14. Zhu H, Hille B, Xu T*. Sensitization of Regulated Exocytosis By Protein Kinase C.
Proc Natl Acad Sci U S A. 99(26): 17055-17059. 2002.
15. Xu T, Bajjalieh SM. SV2 Modulates The Size of The Readily Releasable Pool of
Secretory Vesicles. Nat Cell Biol. 3(8): 691-698. 2001.
16. Xu T, Rammner B, Margittai M, Artalejo AR, Neher E, Jahn R. Inhibition of
SNARE Complex Assembly Differentially Affects Kinetic Components of
Exocytosis. Cell. 99(7): 713-722. 1999.
17. Xu T, Ashery U, Burgoyne RD, Neher E. Early Requirement For Alpha-SNAP And
NSF In The Secretory Cascade In Chromaffin Cells. EMBO J. 18(12): 3293-304.
1999.
18. Xu T, Binz T, Niemann H, Neher E. Multiple Kinetic Components of Exocytosis
Distinguished By Neurotoxin Sensitivity. Nat Neurosci. 1(3): 192-200. 1998.

ABSTRACT
Sequential Activation of Orai1 Subunits by Different Numbers of STIM1
Leads to Multistate Opening of the CRAC Channel
Zhengzheng Li 1, Lin Liu 2, Yongqiang Deng 1, Wei Ji 1, Wen Du 1, Pingyong Xu 1 Liangyi
Chen 1, and Tao Xu 1,2
1National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese
Academy of Sciences, Beijing 100101, P. R. China.
2College of Life Science and Technology, Huazhong University of Science and
Technology, Wuhan 430074, P. R. China.
STIM1 and Orai1 are essential proteins of the Ca 2+ -release-activated Ca 2+ (CRAC)
channel which mediates store-operated Ca 2+ entry. STIM1 is localized at the
endoplasmic reticulum (ER) and functions as the sensor of ER luminal Ca 2+
concentration, and Orai1 is the channel pore subunit at the plasma membrane. STIM1
aggregates underneath the plasma membrane after store depletion and activates the
CRAC channel by directly binding to Orai1. It remains a challenge to determine how
many STIM1 molecules are required to open a CRAC channel. Here, by linking
different numbers of Orai1 subunits with functional cytoplasmic domains of STIM1
(residues 336-485, designated S domains), we provide evidence that each CRAC
channel can be fully activated by eight S domains. Further experiments revealed that
two S domains can fully activate one Orai1 subunit and the CRAC channel can
progressively open as more Orai1 subunits are activated. Our data suggest that full
activation of one CRAC channel complex requires eight STIM1 molecules, and the
activation of the channel is not an “all-or-none” process but undergoes multiple
intermediate states corresponding to different numbers of Orai1 subunits being
activated.

SOLICITED
ABSTRACTS

Chemosensory and Behavior of C.elegans is Regulated by An E.Coli
Noncoding RNA
Jinjing Wu, Jing Ren, Ge Shan
Key Laboratory of Molecular Biophysics, Ministry of Education College of Life Science
and Technology, Huazhong University of Science and Technology, Wuhan 430074,
People’s Republic of China.
It has been known that food as one environmental factor can affect the expression of
genes in animal cells, although we still lack examples of this kind of effect in which the
detailed molecular mechanism is well understood. Here, we found that the expression of
a C. elegans gene, che-2, is down regulated by oxidative stressed E. coli. We
demonstrated that this down regulation is mediated by an E. coli noncoding RNA, OxyS
RNA. Oxidative stress induces the expression of OxyS in E. coli, which through a
mechanism of feeding RNA interference, suppresses the expression of che-2 in C.
elegans with a 17nt complementary sequence between OxyS and che-2 mRNA.
Agonate and Dicer, Key components in the microRNA/ RNA interference pathway in C.
elegans were shown to be required for this regulation of gene expression by E. coli
OxyS RNA. Behavior analysis indicated that oxidative stressed E. coli "utilizes" OxyS
noncoding RNA as a "repellent" to protect itself from bacterial forager such as C.
elegans. These results indicate a co-evolutionary relationship between C. elegans
and a natural food of this animal, E. coli.

Implication of the env Gene of the Human Endogenous Retrovirus W
Family in the Expression of BDNF and DRD3
Shan Li, Honglian Yu, Lijuan Liu, Wang Zheng, Chunlan Liu, Jie Zeng, Yabing Wang,
and Fan Zhu*
School of Basic Medical Science, Wuhan University
It has been reported that HERVs play functional roles in some diseases, such as tumor,
some types of schizophrenia, bipolar disorder, and so on. However, how HERVs
contribute to these diseases is far from clear. We found that HERV-W env could
activate BDNF pathways that might result in the development of schizophrenian. In this
study, U251 Cells were used to study whether HERV env protein could active the
BDNF pathways on the molecular level.

Microvavle Immobilization of Intact C. Elegans for in vivo Calcium
Imaging of Neuronal Activities in Response to External Stimuli
Jingjing Wang, Xiaojun Feng, Wei Du, Bi-Feng Liu*
Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for
Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory,
Department of Systems Biology, College of Life Science and Technology, Huazhong
University of Science and Technology, Wuhan 430074, People’s Republic of China.
Aim To immobilize intact young adult C. elegans for imaging and explore neuronal
activities in response to the changing environments. Methods We adopted a harmless
and innocuous method to immobilize awake behaving C. elegans in a microfluidic
device for imaging in vivo neuronal responses to controllable drug stimuli. A
comb-shaped microfluidic valve was developed for the immobilization of living C.
elegans with minimal influence on its activity for a long period of time. Results Our
microfluidic device achieved the complete immobilization effect of living C. elegans,
similar to the glue-based method. In coupling to a single-drug delivery system, we
investigated the neuronal response and adaptation of the amphid polymodal ASH
sensory neurons to repeated rapid, sharp stimuli such as high concentrations of glycerol.
With a multi-drug delivery system, we sequentially stimulated the same animal with
three different chemical reagents, resulting in successful differentiation between
mutants and wild type. Conclusion The PDMS comb-shaped microvalve demonstrates
the complete immobilization of awake behaving C. elegans. Microfluidic devices also
provide micrometer-sized scales for rapid and sharp change in microenvironment to
observe neuron responses to controllable drug stimuli. Thus, sensitive imaging of
neuronal activities can be conducted in vivo. We expect that microfluidic chips can be
used to study different aspects of nervous system in C. elegans.
Key Words: C. elegans, microfluidic, comb-shaped valve, neuronal activities
*Corresponding author
Email: bfliu@mail.hust.edu.cn Tel: 86-27-87792203

Inhibitory Effect of Ethanol on Gustatory Plasticity in C. Elegans
Mediated by the Serotonin Pathway
Ying Wang, Lichun Tang, Wei Du, Bi-Feng Liu*
Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for
Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory,
Department of Systems Biology, College of Life Science and Technology, Huazhong
University of Science and Technology, Wuhan 430074, People’s Republic of China.
Aim Ethanol can affect the formation of learning and memory in many species.
However, the molecular mechanisms underlying the behavioral effects of ethanol are
still poorly understood. The results from the invertebrate model organisms contribute to
and accelerate the research in higher animal species. So the effect of acute ethanol
exposure on learning and memory in invertebrates yet needs to be studied more in-depth.
Methods In C. elegans, gustatory plasticity is a simple learning paradigm, in which
animals after prolonged pre-exposure to a chemo-attractive salt show chemo-aversion to
this salt. We are using this learning model to study the effect of ethanol on learning and
memory in C. elegans. Results Our results show that ethanol exhibits an acute
inhibitory effect on gustatory plasticity in well-fed C. elegans. Genetic analysis revealed
that mutant animals with defects in serotonin synthetic enzyme tph-1 failed to exhibit
ethanol’s inhibitory effect on gustatory plasticity, however this inhibitory effect could
be restored by expression of tph-1 in the NSM neurons. In addition, tph-1 acts as a
partial downstream target of the BK potassium channel slo-1, regulating acute
intoxicating effects of ethanol in locomotion and egg-laying. Furthermore, we show that
G protein-coupled receptors of serotonin SER-4, SER-7 as well as G-protein α subunits
GPA-3, GPA-11 possibly function in the same genetic pathway to modulate this
response to ethanol. Results of calcium imaging indicate that ethanol directly changes
the activity of ASER neuron that plays a dominant role in chemotaxis to salt.
Conclusion Together, our results demonstrate the distinct role of serotonin pathway in
modulation of acute response to ethanol in gustatory plasticity in C. elegans. Also, the
effect of ethanol on gustatory plasticity explored in our study is a novel example of the
effect of ethanol on learning behavior.
Key Words: Caenorhabditis elegans, ethanol, gustatory plasticity, serotonin pathway
*Corresponding author
Tel: 86-27-87792203 , Email: bfliu@mail.hust.edu.cn

Isoform-Specific Prolongation of KCNQ (Kv7) Potassium Channel
Opening Mediated by a New Drug-Channel Binding Site
Zhaobing Gao1, 5, Tangzhi Zhang2, Meng Wu1, Qiaojie Xiong1, 3, Haiyan Sun1, 4,
Yinan Zhang2, Liansuo Zu2, Wei Wang2,* and Min Li1,*
1Department of Neuroscience, High Throughput Biology Center and Johns Hopkins
Ion Channel Center, School of Medicine, Johns Hopkins University, 733 North
Broadway, Baltimore, MD 21205, 2Department of Chemistry & Chemical Biology,
University of New Mexico, Albuquerque, NM 87131, 3 Current address: Cold Spring
Harbor Laboratory, One Bungtown Road, Cold Spring Harbor, NY 11724, 4 Current
address: Corning Inc. One Science Center Road, Corning, NY 14831, 5 Current address:
Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences,
Chinese Academy of Sciences, Shanghai 201203
Kv7 channels, especially Kv7.2 (KCNQ2) and Kv7.3 (KCNQ3), are key determinants
for membrane excitability. Some chemical modulators for KCNQ are being approved
for therapeutic use as anti-epileptic drugs, such as retigabine ((D-23129,
N-(2-amino-4-(4-fluorobenzylamino)-phenyl). Therefore, a better understanding of
Compound-channel interaction is an area of intense interest. Of particular interest, a
single residue change of W236L in KCNQ2 abolishes its sensitivity to Retigabine. We
have conducted a screen of 20,000 compounds for KCNQ potentiators using rubidium
flux combined with atomic absorption spectrometry (AAS). Here, we report
characterization of a series of new structures with structural similarity to ICA-27243.
We found that they display isoform specificity and induce a marked reduction of
deactivation distinct from that of retigabine. Furthermore, KCNQ2(W236L) remains
fully sensitive to these compounds. This result together with mutagenesis studies and
analyses of combinatorial drug treatments suggests ZTZ compounds confers a new
mode of action presumably by recognizing a new site on channel protein. Our results
argue for a rationale for a deliberate search for channel potentiators with preferential
effects of inhibiting channel close.
Key Words: Potassium Channel, KCNQ, Activator, High Throughput Screening
* To whom the correspondence should be addressed:
Wei Wang, Ph.D., Department of Chemistry & Chemical Biology, University of New
Mexico, Albuquerque, NM 87131. 505-277-2609 (fax). wwang@unm.edu
Min Li, Ph.D., Department of Neuroscience, Johns Hopkins University School of
Medicine, BRB311, 733 North Broadway, Baltimore, MD 21205. 410-614-1001 (fax).
minli@jhmi.edu

Novel USH2A Compound Heterozygous Mutations Cause RP/USH2 in a
Chinese Family
Xiaowen Liu,1,2 Zhaohui Tang,2 Chang Li,2 Kangjuan Yang,3 Guanqi Gan,2 Zibo
Zhang,3 Jingyu Liu,2 Fagang Jiang,1 Qing Wang,2 and Mugen Liu2
1The Union Hospital, Huazhong University of Science and Technology, Wuhan, Hubei,
P.R. China, 2Key Laboratory of Molecular Biophysics of Ministry of Education, College
of Life Science and Technology, Center for Human Genome Research, Huazhong
University of Science and Technology, Wuhan, China, 3Departments of Cell Biology
and Medical Genetics, Yanbian University, Yanji, China
Purpose: To identify the disease-causing gene in a four-generation Chinese family
affected with retinitis pigmentosa (RP). Methods: Linkage analysis was performed with
a panel of microsatellite markers flanking the candidate genetic loci of RP. These loci
included 38 known RP genes. The complete coding region and exon-intron boundaries
of Usher syndrome 2A (USH2A) were sequenced with the proband DNA to screen the
disease-causing gene mutation. Restriction fragment length polymorphism (RFLP)
analysis and direct DNA sequence analysis were done to demonstrate co-segregation of
the USH2A mutations with the family disease. One hundred normal controls were used
without the mutations. Results: The disease-causing gene in this Chinese family was
linked to the USH2A locus on chromosome 1q41. Direct DNA sequence analysis of
USH2A identified two novel mutations in the patients: one missense mutation
p.G1734R in exon 26 and a splice site mutation, IVS32+1G>A, which was found in the
donor site of intron 32 of USH2A. Neither the p.G1734R nor the IVS32+1G>A
mutation was found in the unaffected family members or the 100 normal controls. One
patient with a homozygous mutation displayed only RP symptoms until now, while
three patients with compound heterozygous mutations in the family of study showed
both RP and hearing impairment. Conclusions: This study identified two novel
mutations: p.G1734R and IVS32+1G>A of USH2A in a four-generation Chinese RP
family. In this study, the heterozygous mutation and the homozygous mutation in
USH2A may cause Usher syndrome Type II or RP, respectively. These two mutations
expand the mutant spectrum of USH2A.
Correspondence to:
Dr. Mugen Liu, Huazhong University of Science and Technology Human Genome
Research Center, Wuhan, Hubei, 430074, P.R. China; Phone: 86-27-87794549; FAX:
86-27-87794549; email: lium@mail.hust.edu.cn

UNC-31/CAPS Docks and Primes Dense Core Vesicles in C. elegans
Neurons
Xian-Guang Lin, Min Ming, Mao-Rong Chen, Wei-Pin Niu, Yong-Deng Zhang, Bei Liu,
Ya-Ming Jiu, Jun-Wei Yu, Tao Xu,* and Zheng-Xing Wu*
Key Laboratory of Molecular Biophysics of Ministry of Education, School of Life
Science and Technology, Huazhong University of Science and Technology, Wuhan,
430074 China
UNC-31 or its mammalian homologue, Ca 2+ -dependent activator protein for secretion
(CAPS), is indispensable for exocytosis of dense core vesicle (DCV) and synaptic
vesicle (SV). From N- to the C-terminus, CAPS contains putative functional domains,
including dynactin 1 binding domain (DBD), C2, PH, (M)UNC-13 homology domain
(MHD) and DCV binding domain (DCVBD), the last four we examined in this study.
We employed UNC-31 null mutant C. elegans worms to examine whether UNC-31
functions could be rescued by ectopic expression of full-length UNC-31 vs each of
these four domain-deleted mutants. Full length UNC-31 cDNA rescued the phenotypes
of C. elegans null mutants in response to Ca 2+ -elevation in ALA neurons. Surprisingly,
MHD deletion also rescued UNC-31 exocytotic function in part because the relatively
high Ca 2+ level (pre-flash Ca 2+ was 450 nM) used in the capacitance study could bypass
the MHD defect. Nonetheless, the three other domain-truncation cDNAs had almost no
rescue on Ca 2+ evoked secretion. Importantly, this genetic null mutant rescue strategy
enabled physiological studies at levels of whole organism to single cells, such as
locomotion assay, pharmacological study of neurotransmission at neuromuscular
junction, in vivo neuropeptide release measurement and analysis of vesicular docking,
Using this battery of physiological assays, we found that all four UNC-31 domains are
independently indispensible for UNC-31 to effect its function. The rescue of the MHD
deletion was an in vitro but non-physiological phenomenon which was not recapitulated
in the physiological assays Our results suggest that each of these UNC-31 domains
support distinct sequential molecular actions of UNC-31 in vesicular exocytosis,
including steps in vesicle tethering and docking that bridge vesicle with plasma
membrane, and subsequently priming vesicle by initiating the formation of soluble
N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) core complex.

Cordycepin Protects Against Brain ischemical Reperfusion Injury in
Mice by Inhibition of Matrix Metalloproteinases-3 Expression.
Zhenyong Cheng , Lianjun Guo*,Qing Lv, Xulin Xu
Department of Pharmacology ,Tongji medical college,Huazhong University of Science
and Technology, WuHan, China 430030
Aim: the neuronal death following transient global ischemia - reperfusion in the mouse
takes to occur, Since matrix metalloproteinase-3 (MMP-3) enhances inflammation and
promotes apoptosis in ischemia – reperfusion. providing a potential neuroprotective
effect of cordycepin. Methods: Kun ming male mice (18-25g) were randomly divided
into five groups:control group, brain ischemic- reperfusion injury group, cordycepin
10mg/kg, 20mg/kg and extract of cordyceps militaris(CME) treatmented groups. The
bilateral common carotid artery was occlusion (2VO) for 15 min, and then subjected to
a subsequent 4h period of ischemia reperfusion. The brain protein was extracted and
the expression of matrix metalloproteinases-3(MMP-3) was measured by western blot
analysis. Results: The expression of MMP-3 in the cordycepin- treated group(10mg/kg)
and CME group were decreased by 45.8% and 41.6% contrast with model
group(P

Calcium Signaling in Astrocytes Induced by Photostimulation with
Femtosecond Laser
Yuan Zhao, Yuan Zhang, Wei Zhou, Shaoqun Zeng*
Britton Chance Center for Biomedical Photonics
Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and
Technology,Wuhan, 430074, China
Astrocytes have been proved to actively contribute to brain functions, communicating
with neurons and other brain cells. However, conventional stimulation approaches are
hard to be feasibly utilized to precisely activate astrocytes for delicate investigations.
Here, we developed non-invasive photostimulation with high accuracy to evoke calcium
(Ca 2+ ) signaling in astrocytes. The near infrared (800 nm) femtosecond laser was
focused onto the cell membrane of an astrocyte. Ca 2+ elevation was immediately evoked
in the stimulated cell and subsequently in surrounding cells, forming a radial Ca 2+ wave
throughout the network. Localized transient photoporation was demonstrated to be
induced on the cell membrane by laser irradiation. Extracellular Ca 2+ entry into the
cytoplasm through this tiny poration was necessary for astrocytic Ca 2+ signaling.
Repetitive photostimulation was actually performed on one cell with well-controlled
laser intensity and targeting, resulting in similar responses. Further, different patterns of
Ca 2+ elevation in the stimulated astrocyte were observed by varying femtosecond laser
power. Generally, the amplitude of Ca 2+ response in the stimulated cell was increased
with enhanced stimulating laser power, and concomitantly, the Ca 2+ wave was widened.
Therefore, noncontact photostimulation of astrocytes with femtosecond laser is
demonstrated to be non-disruptive, reproducible, and with high spatiotemporal precision.
Photogenerated Ca 2+ response follows a femtosecond laser-power-dependent manner, so
that distinct Ca 2+ signaling ranges can be feasibly provided for specific studies. This
versatile tool is thus promisingly efficient and helpful for resolving problems that
conventional stimulations are difficult to touch, especially for in vivo studies.
Key Words: astrocyte, Ca 2+ signaling, photostimulation, femtosecond laser,
photoporation
*Corresponding author : Email: sqzeng@mail.hust.edu.cn

ZD7288 Inhibits the Induction Of LTP in an NMDA Independent
Manner at Hippocampal Schaffer Collateral-CA1 Synapses
Wei He, Zhenyong Cheng, Shibin Li, Xulin Xu, Qing Lu, Lianjun Guo*
Department of Pharmacology, Tongji Medical College of Huazhong University of
Science and Technology, Wuhan, 430030, China
Aim: To investigate the effect of ZD7288 on synaptic transmission and high frequency
stimulation (HFS)-induced long-term potentiation(LTP) in the Schaffer collateral-CA1
synapse of mouse hippocampal slices, and identify whether the effect of ZD7288 on the
induction of LTP can be explained by inhibition of N-methyl-D-aspartate (NMDA) type
of glutamate receptors. Methods: Extracellular field excitatory postsynaptic
potentials(fEPSPs) recordings were used to examine the effect of ZD7288 on basal
synaptic transmission and HFS-induced LTP in the Schaffer collateral-CA1 synapse of
mouse hippocampal slices, and measured the evoked depolarisations that by direct
agonist NMDA receptors. Result: Bath application of 10 µM ZD7288 had no effect on
basal synaptic transmission but inhibited the induction of HFS-induced LTP. In addition,
10 µM ZD7288 had no effect on the responses to exogenous NMDA. Conclusion: The
results suggested that ZD7288 has the ability to prevent the induction of LTP at the
Schaffer collateral-CA1 synapse of mouse hippocampus in vitro, and this inhibitory
effect is an NMDA receptor independent manner.
Key Words: ZD7288; synaptic transmission; long-term potentiation; NMDA
Acknowledgments: This work was supported by grants from the National Natural
Science Foundation of China (NSFC, No. 30772559) to Lianjun Guo
First author : Wei He Tel: +86 27 83691762,E-mail: hewei8286291@yahoo.com.cn
* Correspondence author :Tel: +86 27 83691763, E-mail: gljyl@yahoo.com.cn

Isolation, Characterization and Anti-Cancer Activity of SK84, a Novel
Glycine-Rich Antimicrobial Peptide from Drosophila Virilis
Jie Lu*, Mu-ya Shu, Ming-lei Leng, Zheng-wang Chen
Institute of Biophysical & Biochemistry, College of Life Science and
Technology,Huazhong University of Science & Technology, Wuhan, 430074, China
We report herein the isolation and characterization of a novel glycine-rich antimicrobial
peptide purified from the larvae of Drosophila virilis. A range of chromatographic
methods was used for isolation and its antibacterial activity against Bacillus subtilis was
employed to screen for the most active fractions. The peptide, termed SK84 due to its
N-terminal serine, C-terminal lysine and a total of 84 residues, was completed
sequenced using RT-PCR cDNA cloning. SK84 contains a high level of glycine (15.5%)
and a hexaglycine cluster motif in the N-terminal part. SK84 displayed antibacterial
activity against the tested Gram-positive bacteria (Bacillus subtilis, Bacillus
thuringiensis and Staphylococcus aureus), but had no effect on Gram-negative bacteria
(Pseudomonas aeruginosa, Escherichia coli) and fungi (Saccharomyces cerevisiae,
Candida albicans). SK84 had specific inhibitory effects on the proliferation of several
cancer cell lines (Human leukemia THP-1, liver cancer HepG2, and breast cancer
MCF-7 cells), but no hemolytic activity. The results from scanning electron microscopy
observations revealed that SK84 killed THP-1 cells by destroying the cell membranes.
Alignment results show that SK84 is a mature protein processed from the pseudoprotein
GJ19999 from Drosophila virilis, and is very similar to several pseudoproteins from
different Drosophila species. SK84 induces the apoptosis of HepG2 cells by stimulating
the caspase-3 activation. Our results show that SK84 represents a novel glycine-rich
peptide family in Drosophila species with antimicrobial and anti-cancer cell activities.
Key Words:antimicrobial peptide; glycine-rich peptide; Drosophila virilis;
anti-cancer-cell activity;apoptosis.
*Corresponding author to:
Tel: 86-18971142265;Fax: 86-027-87792024;E-mail:lujie.jane@163.com

Foamy Virus as Potential Vectors for Gene Therapy in Nervous Disorders
and Signal Pathway in Neural Cell Transduction
Ying-Ying ZHANG#, Yong-Juan LIU#, Guo-Guo ZHU, Yan-Yan QIU, Biwen PENG,
Jun YIN, Wan-Hong LIU* and Xiao-Hua HE*
School of Medicine, Wuhan University, Wuhan 430071, China
# Equal contribution to this work
Foamy viruses are a type of retrovirus with a wide range of hosts and non-pathogenicity.
Due to their unique structures, safety and the large packaging capacities, foamy viruses
have been developed into an effective vector system for gene therapy. However, it is
still a controversial issue whether foamy virus vectors could be as exogenous gene
conveyance in the nervous system located a large number of growth-arrested cells. In
this review, we focus on this issue and analyze the reasons for their potentiality of
infecting the growth-arrested cells. Several evidences in papers (from 1997 to 2009)
concerning transduction of the growth-arrested cells and neurons by foamy virus vectors
were compared and analyzed. It is demonstrated that foamy virus vectors can transduce
some growth-arrested cells and neurons. Assuredly, further proofs are needed in the
future. However, these facts suggested that foamy viruses may become potential
transfer vectors for gene therapy in nervous disorders and even for signal pathway in
neural cell transduction.
Key Words: foamy virus vectors; growth-arrested cells; neurons; gene therapy;
neuro-signalling transduction
Acknowledgment: This work was supported by National Natural Sciences Foundation
of China (No. 30870856 and 30970145), Program for New Century Excellent Talents,
Wuhan University (No. NCET-07-0630),the Scientific Research Foundation for the
Returned Overseas Chinese Scholars in State Education Ministry, Research Fund for the
Doctoral Program of Higher Education of China (No. 20090141110010) and the
Independent Scientific Research Foundation of Wuhan University (No.3081002).
*Corresponding author
Tel: 86-27-68759985, Fax: 86-27-68759991.
E-mail: liuwanhong@whu.edu.cn or hexiaohua@whu.edu.cn

Role of Heat on the Expression and Function of KCNQ2
Fang Yu#, Yuncui Wang#, Yanlan Liu, Wanhong Liu, Biwen Peng* and Xiaohua,He*
School of Medicine, Wuhan University, Wuhan 430071, China
# Equal contribution to this work
Objective: KCNQ2 (Kv7.2), Voltage-dependent potassium channels, plays a key role in
regulating neuronal excitability. Mutations in neuronal KCNQ2/3 subunits, the
molecular correlates of M current, have previously been linked with benign familial
neonatal epilepsy (BFNC). In this study, we measured the temperature sensitivity of
KCNQ2 channel and determined whether temperature affected the excitability of the
nervous system by the regulation of KCNQ2 channels.Methods: The wild type and
mutant cDNAs of human KCNQ2, R214W and Y284C genes were subcloned into
pEGFP-C1 plasmid, and then were transfected into 293T cells respectively. The
KCNQ2 protein expression at different temperature was detected by fluorescence
confocal microscopy and Western-blotting. While whole-cell patch clamp was
performed to record the effect of temperature electrophysiological activity of KCNQ2
potassium channel. Results: Total fluorescence intensity values of wtKCNQ2,R214W
and Y284C were enhanced with the temperature increasing from 37℃ to 40 ℃
(P

Possible Involvement of TRPV1 in neonatal Febrile Seizure Mice
Yuan Xiao-ran Yu Hong-mei He Xiao-hua and Peng Bi-wen
Department of physiology, School of Basic Medical Science, Wuhan University
Febrile seizures (FSs) are acute symptomatic seizures that occur in response to a fever
in an age-specific manner. Although FS are largely benign, complex FS increase the
risk to develop temporal lobe epilepsy (TLE). TRPV1 is a capsaicin/vanilloid-gated
cation channel expressed predominantly by primary nociceptive neurons and can
alternatively be activated by elevated temperatures (>43℃). Studies involving systemic
or intrahypothalamic capsaicin administration have suggested a role for TRPV1 in body
temperature control. To explore whether TRPV1 is involved in febrile seizure in
immature brain, behavioral responses were observed on neonatal C57BL6/J mice and
TRPV1 -/- mice. Hyperthermia was induced by warm air in 14-day-old mice. P14 pups in
TRPV1 -/- group showed prolonged latency, shorten duration and lower degree of
seizures compared to wild type mice. Y-maze and Open field test did not show much
difference in two groups at the age of P30. Western-blot and immunohistochemistry
were also used to find out the different expression of TRPV1 between the control group
and FS-model group in wild type mice. These results suggest that TRPV1 probably
participate in the generation of fever seizure and thus may provide new leads for
treatment of children at risk for complex FS and TLE.
Key Words: TRPV1; Febrile Seizure; behavior response; neonatal
Acknowledgment: This work was supported by National Natural Sciences Foundation
of China (No. 30970994 and 30770734), Program for New Century Excellent Talents,
Wuhan University (No. NCET-07-0630),the Scientific Research Foundation for the
Returned Overseas Chinese Scholars in State Education Ministry, and Research Fund
for the Doctoral Program of Higher Education of China (No. 20090141110010).
*Corresponding author: Email: pengbiwen@whu.edu.cn

The Kv1.3 Channel Blocker Scorpion Polypeptide has an Therapeutic
effect on EAE Rats with a Molecule Mechanism
Zhi Li1#, Wanhong Liu1#, Song Han2, Liang Li1, Yuanteng Fan1, Zhihao Wang1, Bo
Yang1, Wu Yingliang2*, Xiaohua He1* and Wenxin Li2
1School of Medicine, Wuhan University, Wuhan 430071, China
2College of Life sciences, Wuhan University, Wuhan 430072, China
# Equal contribution to this work
Objective: Experimental autoimmune encephalomyelitis(EAE), an ideal model of
human multiple sclerosis(MS), is a kind of autoimmune disease in central nervous
system .We are to investigate the efficacy of the Kv1.3 channel blocker scorpion
polypeptide on the inhibition of T cell proliferation in vitro and the therapy of EAE rats
in vivo .Furthermore, to study the T cell activation signaling transduction pathways
which are associated with the Kv1.3 channel.Method: Female inbred Sprague
Dawley(SD) rats with age of 8-10weeks were immunized by their own cerebral and
spinal cord homogenate in Freund's complete adjuvant with subcutaneous injection of
0.06g/100g antigen. The administration of polypeptide was 100ug/kg .i.p. for 14days,
rats were observed for 16days and the clinical score of EAE were recorded. The lesion
tissues were Hematoxylin -Eosin(H.E.) stained to observe the inflammatory infiltration
under the microscope. By transmission electron microscopy (TEM),the structure of
loose myelin and the axonal damage of neurons were observed. The cytokine levels of
IL-2,IFN-γ,TNF-α from the rats serum were detected by the ELISA assay. In vitro T
cells proliferation assay, spleen cells from EAE rats were cultured in a 96-well plates
and activated by antigen, treated with different doses of drugs, cells were cultured for
72h and were added with [ 3 H]-thymidine for 18h before being harvested , the cpm
values(the [ 3 H]-thymidine incorporation )were measured by the liquid
scintillation .Result: From the H.E. sections , the polypeptide can efficiently attenuate
the inflammatory infiltration into the central nervous system. From the ultra structure of
the tissues, the myelin and axon from polypeptide treatment group were basically
normal.IL-2 and TNF-α of EAE group were significantly higher than the control
(* P

pathways, and the change of signal molecules after the intervention of scorpion
polypeptides.
Key Words: EAE , Kv1.3 channel, T cells, signaling pathways
Acknowledgment: This work was supported by Program for New Century Excellent
Talents, Wuhan University (No. NCET-07-0630),the Scientific Research Foundation for
the Returned Overseas Chinese Scholars in State Education Ministry, and Research
Fund for the Doctoral Program of Higher Education of China (No. 20090141110010).
*Corresponding author
E-mail: hexiaohua@whu.edu.cn or liwxlab@whu.edu.cn

IGF-1 Promote BPH-1 Cell Proliferation Primarily via Activating
mTOR-dependent Translational Increases in Cyclin D Proteins
Ke Gong, Chao Chen and Wenhua Li*
Department of Cell Biology, College of Life Sciences, Wuhan University, Wuhan
430072, PR China.
Aim: Insulin-like growth factor-1 (IGF-1) and its binding protein (IGFBP-3) play a
crucial role in the initiation and progression of prostate and benign prostatic hyperplasia
(BPH). We have previously report that stromally expressed c-Jun protein modulates the
epithelial proliferation via regulating production and paracrine signals of IGF-1, which
promote the BPH-1 cells proliferation through upregulate cyclin D protein levels.
Methods: Western blots analyses for protein levels and cell cycle array of FACS were
performed. Results: In this study, the exact molecular mechanisms of these regulations
were investigated. We found that IGF-1 increases cyclin D proteins and promotes cell
cycle transition from G0 to S phase, bringing in cell proliferation. IGF-1 almost didn’t
affects cyclin D mRNA levels and it primarily stimulates post-transcriptional increases
in cyclin D proteins. IGF-1 stimulation increases cyclin D protein in translation levels
via activation mammalian target of rapamycin (mTOR) pathway in BPH-1 cells. Two
vital factors of mTOR-downstream, the eukaryotic translation initiation factor-4E
binding protein-1 (4EBP1) and 70-kD ribosomal protein S6 kinase (p70S6K), were
activated after IGF-1 treatment. Blocking mTOR will decrease cyclin D levels and
offset IGF-1 stimulation. Moreover, we found that, in BPH-1cells, activated mTOR by
IGF-1 increases cyclin D protein expression is mediated through activation of the
PI3K/Akt pathways cells, but not MAP kinase pathway. Also, our data indicate that
IGF-1 probably influences the combination of cyclin D and ubiquitin, and then stabilize
the cyclin D protein. Conclution: These results elucidate the molecular transduction
pathways of IGF-1 promote BPH-1 cell proliferation, which will be help to improve our
understanding of BPH and develop new clinical therapy for it.
Key Words: IGF-1; BPH; mTOR; cyclinD
*Corresponding Authors:
Wenhua Li, Email: whli@whu.wdu.cn, Tel: 86-27-68756711

GABAB Receptor-Mediated Rap1 Activation and GB1/Rap1GTP
Interaction in Neuronal System
Zongyong Zhang, Siluo Huang,Ninghua Sun,Lin Wu,Yilei Zhang,Jianfeng Liu, Li Su
Sino-France Laboratory for Drug Screening, Key Laboratory of Molecular Biophysics of
Ministry of Education, School of Life Science and Technology, Huazhong University of
Science and Technology, Wuhan, 430074 ,China
Rap1, a Ras-related small GTPase, binds to either GTP(Rap1GTP active form ) or GDP
(Rap1GDP inactive form) to regulate cell proliferation, adhesion, or migration.
Gamma-aminobutyric acid (GABA) B receptors, which are GB1/GB2 heterodimer, play
an important role in many physiological activities in neuron. We found that activation
of GABAB receptor stimulated with Baclofen or CGP7930, analogs of GABA, activated
Rap1 signaling both in cultured mice cerebellar granule neurons (CGNs) and in
HEK293 cell. This activation can be blocked by depleting of Ca 2+ in the medium with
Ca 2+ chelator BAPTA. We further found the active Rap1 translocated to premembrane
region and interacted directly with GB1 C-terminal via its hypothetical inter-protein
β-sheet structure.
Key Words : Rap1, GABA, GABAB receptor, protein-protein interaction

Therapeutic Effect of Flupirtine on RepetitiveFebrile Seizures Rats
Yanlan Liu, Fang Yu, Biwen Peng, Yuncui Wang, Wanhong Liu, Xiaohua,He*
School of Medicine, Wuhan University, Wuhan 430071, China
Aim:KCNQ2 and KCNQ3 encode subunits of neuronal M-type K + channels,which are
key regulators of brain excitability.This study tests whether Flupirtine , a selective
KCNQ2 activator ,is effective in the experimental model of repetitive febrile seizures.
Methods: Our study includes 40 male Sprague-Dawley (SD) rats at postnatal day 10
(P10) divided into normal、febrile seizures、phenobarbital and flupirtine groups. We
induced repetitive febrile seizures starting at postnatal day 10 (P10) with hot water
bath(45.0 ℃ ) for 8 consecutive days. In phenobarbital and flupirtine groups,
phenobarbital and flupirtine at the dose 30mg/kg b.m. was administered respectively 2
hours before febrile seizures . febrile seizures group and normal group rats received
only normal saline. At P28, we induced febrile seizures again to test the rats'
susceptivity to febrile seizures. At P30, the Morris water maze were applied to measure
the rats’ learning and memory abilities in each group. Thereafter, hippocampal slices
were prepared for histological and morphometric examination.
Results: Both Flupirtine and phenobarbital can significantly increase the latency and
decrease the rate and duration of febrile seizures when administered before FS(P

Rapid Detection of Deletion of the PMP22 Gene from a Kindred in China
with Hereditary Neuropathy with Liability to Pressure Palsy by Real-time
Quantitative PCR
Yuan He1, Qiang Wu2, Qianqian Wang1, Wanhong Liu1* and Xiaohua He1*
1School of Medicine, Wuhan University, Wuhan 430071, China
2Wuhan General Hospital, Wuhan 430072, China
The hereditary neuropathy with liability to pressure palsy (HNPP) is an
autosomal-dominantly inherited peripheral neuropathy, and the deletion of the
peripheral myelin protein (PMP22) gene in chromosome 17p11.2-12 is the main cause
for it, accounting for approximately 70% of all cases. In this study we reported the
clinical phenotypes and electrophysiological findings from one patient in a kindred in
china who was affected by HNPP. At the same time, the deletion of PMP22 gene which
came from two patients and six asymptomatic normal members in the same kindred was
studied by real-time quantitative PCR using SYBR Green I. The copy number of the
PMP22 gene was determined by the comparative threshold cycle method and the
albumin was used as a reference gene. This study disclosed that the PMP22 gene from
two patients did deleted, which was the same as the previous report in other nations, and
six asymptomatic normal members had no deletion except two members. We conclude
the two asymptomatic members with deletion of the PMP22 gene might develop some
relative symptoms one day.
Key Words: HNPP; Peripheral neuropathy; PMP22; Real-time quantitative PCR
Acknowledgment: This work was supported by Program for New Century Excellent
Talents, Wuhan University (No. NCET-07-0630),the Scientific Research Foundation for
the Returned Overseas Chinese Scholars in State Education Ministry, and Research
Fund for the Doctoral Program of Higher Education of China (No. 20090141110010).
*Corresponding author
E-mail: liuwanhong@whu.edu.cn or hexiaohua@whu.edu.cn

Therapeutic Effect of Flupirtine on Repetitive Febrile Seizures Rats
Yanlan Liu, Fang Yu, Biwen Peng, Yuncui Wang, Wanhong Liu, Xiaohua,He*
School of Medicine, Wuhan University, Wuhan 430071, China
Aim:KCNQ2 and KCNQ3 encode subunits of neuronal M-type K + channels,which are
key regulators of brain excitability.This study tests whether Flupirtine , a selective
KCNQ2 activator ,is effective in the experimental model of repetitive febrile seizures.
Methods: Our study includes 40 male Sprague-Dawley (SD) rats at postnatal day 10
(P10) divided into normal、febrile seizures、phenobarbital and flupirtine groups. We
induced repetitive febrile seizures starting at postnatal day 10 (P10) with hot water
bath(45.0 ℃ ) for 8 consecutive days. In phenobarbital and flupirtine groups,
phenobarbital and flupirtine at the dose 30mg/kg b.m. was administered respectively 2
hours before febrile seizures . febrile seizures group and normal group rats received
only normal saline. At P28, we induced febrile seizures again to test the rats'
susceptivity to febrile seizures. At P30, the Morris water maze were applied to measure
the rats’ learning and memory abilities in each group. Thereafter, hippocampal slices
were prepared for histological and morphometric examination.
Results: Both Flupirtine and phenobarbital can significantly increase the latency and
decrease the rate and duration of febrile seizures when administered before FS(P

TEF3(Transcription Enhancer Factor 3)Induced Angiogenesis through
VEGF Pathway
Xin Liu1, Cheng Qiao1, and Dezheng Zhao2,4 Huiyan Zeng2,3,4*,
1.Department of Microbiological and Biochemical Pharmacy, College of Pharmacy ,
Wuhan University, Wuhan 430071, PR China 2 .Departments of Pathologyand
Medicine (Division of Molecular & Vascular Medicine), 3. Center for vascular Biology
Research 4.Beth Israel Deaconess Medical Center and Harvard Medical School, Boston
MA 02215, USA
Aims: Transcription Enhancer Factor-3 (TEF3, RTEF-1, TEAD4) belongs to a family
of transcription factor proteins. Although it has been extensively studied, the signaling
pathways mediating its biological functions during angiogenensis are still not
understood. Recently we identified a novel VEGF targeted gene, Down Syndrome
Candidate Region 1 isoform 1L (DSCR1-1L) that is important for the function of VEGF
in pathological angiogenesis and is expressed in human tumor vasculature, not in
normal vasculature, rendering it a specific target for tumor angiogenesis therapy. TEF3
was required for DSCR1-1L expression through binding to the M-CAT site in its
promoter and could be an attractive target for anti-angiogenesis therapy. It was also
reported that TEF3 expression is induced by hypoxia and TEF3 overexpression
increases VEGF gene transcription and in vitro under hypothexia condition. Ours aims
are to investigate the role of TEF3 in angiogenesis. Methods: We overexpressed TEF3
in HUVEC cell and we used the yeast two-hybrid system to find the proteins interaction
with the TEF3. Results: We reported previously that another DSCR1 isoform,
DSCR1-1L, was also upregulated by VEGF-A 165 in cultured endothelial cells and in
several in vivo models of pathological angiogenesis and that different from DSCR1-4,
DSCR1-1L overexpression alone induced cultured endothelial cell proliferation and
promoted angiogenesis in Matrigel assays. We also demonstrated that TEF3 directly
interacted with the putative M-CAT site in the DSR1-1L promoter in vitro and in vivo.
Finally, Overexpression of TEF3 induces HUVEC proliferation, migration and tube
formation. Conclusion: TEF3 upregulatecd DSCR1-1l expression with the VEGF
stimulation,and TEF3 itself can promote angiogenesis in endothelial cells. Then it was
tested that TEF3 also acts as a novel factor in VEGF-induced angiogenesis signaling
pathway.
Key Words: VEGF; TEF3; angiogenesis
Authors:
Xin Liu ,Email: lx@whu.edu.cn;
Huiyan Zeng, Email: hzeng@caregroup.harvard.edu Tel: 86-27-68759006

Does the Metabotropic Glutamate Receptor 7 (Mglur7) Play a Role in
Schizophrenia?
Yali Zhao, Yu Fan, Qi Xu*, Yan Shen*
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences
Chinese Academy of Medical Sciences and Peking Union Medical College, Tsinghua
University. No. 5, Dong Dan San Tiao, Beijing, China, 100005
Background: The metabotropic glutamate receptor 7 (mGluR7) is a class C G-protein
coupled receptor found in the pre-synaptic elements of various types of neurons, which
plays a critical role in regulating the glutamate transmission. Glutamate is the
predominant excitatory neurotransmitter in the central nervous system. Glutamate
blocking drugs such as phencyclidine can mimic the psychosis symptoms and cognitive
problems associated with schizophrenia, which is a common yet severe psychiatric
disorder and characterized by hallucinations, delusions and cognitive deficits with a
lifetime risk of about 1%. Aim: The purpose of this study is to investigate the
contribution of mGluR7 to schizophrenia susceptibility. We genotyped 14 single
nucleotide polymorphisms (SNPs) located in the functional region of mGluR7 among
549 patients with schizophrenia and 562 controls origining from Chinese utilizing
Sequenom MassARRAY iPLEX Gold platform (Sequenom, San Diego, California).
The differences of allelic and genotypic frequencies between patients and controls were
examined using Unphased software (v 3.0.12). Results: We observed no significant
allelic associations with the disease in any of the 14 SNPs. We also observed no
significant genotypic differences between patients and controls. Conclusion: These
results suggested that mGluR7 may not play a major role in schizophrenia pathogenesis
in the Chinese population as it is a multifactor disease.
Key Words: metabotropic glutamate receptor 7; Glutamate; schizophrenia;
Author: Yali Zhao, Tel: 86-10-65296432; Email: hollyzyl@126.com;
Corresponding author: Qi Xu, Tel: 86-10-65296432; Email: qixu@vip.sina.com

Effects of Yishendaluo Decoction on Axonal Degeneration, Inflammatory
Reaction, and Neurological Function in a Mouse Model of Experimental
Autoimmune Encephalomyelitis
Xiaoling Shang1, Ying Gao2, Ling Yin3, Jintao Zhang4, Shuoren Wang2
1Department of Basic Theory of Traditional Chinese Medicine, Basic Medical College,
Changchun University of Chinese Medicine,Changchun 130117, Jilin Province, China, 2.
Key Laboratory of Chinese Internal Medicine, Ministry of Education, Dongzhimen
Hospital of Beijing University of Chinese Medicine, Beijing, 100700, China, 3.Center for
Neuroinformatics, General Hospital of Chinese PLA, Beijing 100853, China,
4.Department of Neurology, the 88 Hospital of Chinese PLA, Taian 271000, Shandong
Province, China
Aim: To investigate the effects of Yishendaluo decoction on a mouse model of experimental
autoimmune encephalomyelitis. Methods: A total of 96 healthy, female, SJL/J mice, aged 8-12
weeks, were equally and randomly assigned to normal, model, hormone, and Chinese medicine
groups. A total of 0.2 mL antigen preparation, supplemented with 150 μg PLP139-151 and
400 μg H37RA, was subcutaneously injected into the upper abdomen of mice from the model,
hormone, and Chinese medicine groups. Mouse models of experimental autoimmune
encephalomyelitis were established by intravenous injection of 0.1 mL Bordetella pertussis
solution containing 0.6 × 106 Bordetella pertussis at days 1 and 3. Mice from the model,
Chinese medicine, and hormone groups were respectively subjected to 0.2 mL saline, 2 g/kg
Yishendaluo decoction, and 0.078 mg/kg prednisone acetate, once daily for 14 consecutive days.
Mice from the normal group were left intact. Results: A few inflammatory cell infiltration,
nerve fiber breakage and slight demyelination were detected in the central nervous system of
mice from the Chinese medicine and hormone groups compared with the model group.
Expression of β-amyloid precursor protein and p38 protein was significantly diminished in the
central nervous system of mice from the Chinese medicine and hormone groups compared with
the model group (P < 0.05 or P < 0.01), and the decrease was greatest in the Chinese medicine
group. The decrease in mouse weight was not significant, and neurological function scores were
less in the Chinese medicine and hormone groups compared with the model group (P < 0.05 or
P < 0.01). Interferon-γ levels were significantly reduced (P < 0.01), and interleukin-4 levels
were significantly increased (P < 0.01) in the brains of the Chinese medicine and hormone
groups, compared with the model group. Conclusion: Yishendaluo decoction improved
neurological function in mice with experimental autoimmune encephalomyelitis by
downregulating β-amyloid precursor protein expression, resisting axonal degeneration, and
relieving inflammatory reaction. The anti-inflammatory mechanism was regulated by inhibition
of the p38 mitogen-activated protein kinase signal pathway.
Key Words: autoimmune encephalomyelitis; multiple sclerosis; Yishendaluo decoction; β
-amyloid precursor protein

Imaging Neuronal Population in Vitro with Acousto-Optic Deflector
Based Random Access Two-Photon Microscopy
Shaoqun Zeng, Wei R Chen, Qingming Luo
Britton Chance Center for Biomedical Photonics,Wuhan National Laboratory for
Optoelectronics-Huazhong University of Science and Technology, Wuhan 430074,
China
Two-photon microscopy has grown up to be an important technique in biology research
especially in the field of neuroscience for its high penetration depth and
three-dimensional selectivity. However, its imaging rate is limited by the mechanic
scanning mechanism and cannot satisfy the requirement for imaging the encoding
pattern of brunches of dendrite or cell populations in brain tissue. Laser scanning with
two sequential orthogonally oriented (2D) acousto-optical deflectors (AODs) does not
involve actual mechanical movement and thus provides a fast scanning rate, high
precision, and stability. 2D AOD scanning also allows random access to each pixel in
the field of view (FOV). Random scanning in regions of interest can devote dwell time
to pixels of interest and increase the signal-to-noise ratio and the frame-capture rate, and
would provide unique applications in neuroscience research. However this random
scanning two-photon microscopy with femtosecond laser is frustrated by the temporal
and spatial dispersion. With a special dispersion compensation scheme, we have
constructed a random scanning two-photon microscope and are able to track the fast
neuronal activity which is not available with other techniques. In this presentation, we
will show 1) the evolution of the femtosecond laser pulse after passing the AOD
scanner, 2) a technique to compensate the spatial and temporal dispersion
simultaneously and facilitate two-dimensional random access two-photon microscopy,
and 3) a special optimization algorithm which is particularly effective to reconstruct the
firing pattern from calcium signal of low signal to noise ratio. The whole system is
validated with calcium signals recorded from neurons of brain slices.
Key Words: neuronal activity, two-photon microscopy, acousto-optic deflector, firing
pattern
*Corresponding author : Email: sqzeng@mail.hust.edu.cn

Inhibition of Protein Phosphatase 2A Activity Plays a Key Role for Normal
Human Cells to Acquire TRAIL-Sensitive Phenotype during Tumorigenesis
Yansheng Li1, Yajun Xiao1, Xuegang Wang1, Shaoyong Chen2, Siping Zeng1, Changshuai
Shao1, Xiaohui Qian1,Rong Wang1, Xuanyu Chen1, Quansheng Du3, Aria F. Olumi4,
Hongmei Yang5, Xiaoping Zhang1§
1. Department of Urology, Union Hospital, Tongji Medical School, Huazhong University of
Science and Technology, Wuhan 430022, China, 2.Cancer Biology Program,
Hematology-Oncology Division, Department of Medicine, Beth Israel Deaconess Medical
Center, Harvard Medical School, Boston, Massachusetts 02215, USA 3.Cancer Center,
Medical College of Georgia, Augusta, GA 30912, USA4. Department of Urology,
Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
5.Department of Pathogen Biology, Tongji Medical School, Huazhong University of Science
and Technology, Wuhan 430030, China
Aim: Tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) is a promising
anticancer agent because it induces apoptosis in most cancer cells but spares normal cells. The
aim of the study is to determine how normal human cells acquire TRAIL-sensitive phenotype
during the process of malignant transformation. Methods: An experimental cell system was
developed by sequential introduction of hTERT, SV40 LT, SV40 ST and H-Ras into normal
human prostatic epithelial cells. Then these cells were treated with TRAIL by using MTT assay.
After determining which cells became sensitive to TRAIL, the underlying mechanisms were
explored. Results: For the first time, we demonstrate that introduction of SV40 ST into cells
changes TRAIL phenotype from TRAIL-resistant to TRAIL-sensitive, activity inhibition of the
protein phosphatase 2A (PP2A) either by SV40 ST or okadaic acid (OA) sensitizes normal
human prostatic epithelial cells to TRAIL-induced apoptosis during premalignant period of
tumorigenesis and tumor suppressor gene PP2A may exert its antiapoptotic by negatively
regulating c-Fos/AP-1. In addition, low-dose OA treatment sensitizes TRAIL-resistant cancer
cells to TRAIL, which indicates PP2A inhibitors such as OA can potentially used as an
enhancer of apoptosis induced by TRAIL or TRAIL-like agents. Conclusion: Inhibition of
PP2A activity is a key step for normal human cells to acquire TRAIL-sensitive phenotype
during tumorigenesis, and lower PP2A activity might be a major determinant for cancer cells to
be sensitive to TRAIL-induced apoptosis.
Key Words: TRAIL, the protein phosphatase 2A (PP2A), Apoptosis, SV40 ST, Okadaic acid
(OA)
Grant Support: the National Natural Science Foundation of China (NSFC) (30572139),
(30872924) and Program for New Century Excellent Talents in University from Department of
Education of China (NCET-08-0223) to XZ
§ Corresponding Authors:
Xiaoping Zhang:1277 Jiefang Dadao,Wuhan 430022,Hubei Province,China, Department of
Urology, Union Hospital, Email: xiaoping.zhang2008@gmail.com

Phone: 86-27-85351625
A Novel Mutation of PAX3 on a Large Chinese Family with
Waardenburg Syndrome
Juan Liu1, Xiangfu Meng2, Caixia Sun2, Jingmin Wen1, Yong Gao1, Jing Yu Liu1*
1Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life
Science and Technology, Center for Human Genome Research, Huazhong University
of Science and Technology, 430074, Wuan, Hubei, P. R. China;2 People Hospital of
Xiajin, Xiajin, Shandong, 253200, China
Objectives: Waardenburg syndrome (WS) is an autosomal dominant and genetically
heterogeneous disorder characterized by manifests with sensorineural deafness and
pigmentation defects of the hair, skin, and iris. It is classified into four types depending
on the presence or absence of additional symptoms, WS1, WS2, WS3 and WS4. WS1
and 3 are due to mutations in the PAX3 gene. WS2 are associated with mutations of
MITF, SNAI2, and SOX10, in addition, there are two loci, 1p21-p13.3 and 8p23. WS4
can result from mutations in EDNRB and EDN3.Methods: Linkage analysis was used
to identify the chromosomal location of the disease gene, direct DNA sequence analysis
was used for mutation detection and restriction fragment length polymorphism (RFLP)
analysis was checked this mutation in the family. Results: A large five-generation
Chinese family from Shandong Provience with autosomal dominant WS1 have
identified and characterized. Linkage analysis was identified with marker D2S126,
which is located near Pax3 gene. The PAX3 gene is closely linked to the disease of this
family. Mutational analysis of all exons and exon-intron boundaries of PAX3, a novel
mutation c.272A>G in the proband was found. RFLP analysis showed that the mutation
is in all patients, but not in all normal individuals and 200 normal controls. The
mutation occurs at a highly conserved residue in the paired domain (PD) of PAX3,
which is the DNA binding domain. Conclusion: These results indicate that the
c.272A>C mutation of PAX3 gene probably causes WS in the family and expand the
mutation spectrum of PAX3 gene.
Key Words: Waardenburg syndrome, PAX3, Linkage analysis, Mutation analysis
*Correspondence to: Dr. J. Y. Liu, liujy@mail.hust.edu.cn

Identification a Novel MYO6 Mutation Associated with Autosomal
Dominant Non-Syndromic Hearing Impairment in a Large Chinese
Family
Yulei Li1,2, Zhenping Xu1,2, Xiangyang Zhang1, Juan Li1, Jing Yu Liu1*
1Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life
Science and Technology, Center for Human Genome Research, Huazhong University
of Science and Technology, 430074, Wuan, Hubei, P. R. China; 2Department of Life
Science and Technique, Xinxiang Medical University, Xinxiang, 453003, China)
Objectives: Hearing impairment is a disorder of partial or complete hearing loss, it is
caused by organic or functional pathological changes in the human auditory system. Hearing
impairment have an estimated incidence rate of approximately 1 in 500 newborns with bilateral
congenital sensorineural hearing loss≥40dB in developed countries. And the ratio up to 2.7‰
for the age of 5. In the adolescent, this number rises to 3.5‰. Genetic factor is the most
important nosogenesis among all the factors. Most of the inherited cases are non-syndromic,
with the only symptom of auditory dysfunction, not accompanied with other malformation or
disorder. Most of the non-syndromic hearing impairment (NSHI) is caused by monogenic
mutation, and they are genetically heterogeneity. Many autosomal dominant NSHI genes have
been identified. Most of these genes have a high expression or specially expressed in the inner
ear cells. By the molecular basis of these studies, it gives us a further understands of the
physiological mechanism of auditory system and the pathological basis of hearing
impairment.Methods: Linkage analysis was carried out by STR Markers flanking all known
autosomal dominant NSHI genes and direct DNA sequence for the proband was used to
mutation analysis by ABI PRISM 3100 Genetic Analyzer.Results: A large Chinese Han family
with an autosomal dominant non-syndromic hearing impairment have been identified and
characterized. Linkage analysis of genotyping data from the family showed the DFNA22 locus
can not be excluded. There is only one candidate gene MYO6 in this locus which is responsible
for NSHI. Mutational analysis showed a novel missense mutation c.599A>G in exon 8 was
found in proband, it was co-segregated with the affected individuals in the family and did not
exist in the unaffected family members and 100 unrelated normal controls. The mutation results
in a substitution of an asparagine residue by a serine residue at codon 200 (p.N200S). The
p.N200S occurred in the motor domain near the ATP binding site of myosin VI, and the N200
residue is evolutionally highly conserved from flagellum to human.Conclusion: It is presumed
that the mutation p.N200S in the motor domain maybe affect the generation of mechanical force
in myosin VI, and lead to the dysfunction of inner ear hair cell, and eventually result in the
hearing impairment.
Key Words: non-syndromic hearing impairment, mutation analysis, myosin VI, p.N200S,
motor domain
*Correspondence to: Dr. J. Y. Liu, liujy@mail.hust.edu.cn

MAPK Scaffolding by BIT1 in the Golgi Complex Modulates Stress
Resistance
Ping Yi1,2,3*, Duc Thang Nguyên4*, Arisa Higa-Nishiyama1,2, Patrick Auguste2,4,
Marion Bouchecareilh1, Michel Dominguez5, Regula Bielmann4, Sandrine Palcy2,
Jian Feng Liu3 and Eric Chevet1,2,3 ‡
1Avenir, INSERM U889, Bordeaux, France; 2Université Victor Segalen Bordeaux 2, IFR
66, F-33076, Bordeaux, France; 3Key Laboratory of Molecular Biophysics of Ministry of
Education, School of Life Science and Technology, Huazhong University of Science and
Technology, Wuhan, Hubei, China; 4Department of Surgery, McGill University,
Montreal, QC, Canada; 5HyperOmics Farma, Montreal, QC, Canada;
*These authors contributed equally to this work
The endoplasmic reticulum (ER) is an essential organelle whose major functions are to
ensure proper secretory protein folding and trafficking. These mechanisms involve the
activation of specific ER-resident molecular machines, which might be regulated by
their membranous environments. Based on this observation, we aimed to characterize
the proteome of ER-membrane microdomains to identify new components of the ER
that have a role in secretory pathway-associated functions. Using this approach with dog
pancreatic rough microsomes, we found that mitochondrial Bcl-2 inhibitor of
transcription (BIT1) localized in the early secretory pathway and accumulated in the
Golgi complex. Using both a chimeric protein of the luminal and transmembrane
domains of ER-resident TRAPa and the cytosolic domain of BIT1, and silencing of
BIT1 expression, we perturbed endogenous BIT1 oligomerization and localization to
the Golgi. This led to enhanced ERK signaling from the Golgi complex, which resulted
in improved stress resistance. This work provides the first evidence for the existence
of ER microdomains that are involved in the regulation of BIT1 structure and
trafficking, and identifies BIT1 as a negative regulator of the ERK-MAPK signaling
pathway in the Golgi.
Key Words: BIT1, MAPK, Secretory pathway, Stress
‡Author for correspondence: Eric Chevet, eric.chevet@u-bordeaux2.fr

Identification of a Novel Genetic Locus on Chromosome 8p21.1-q11.23
for Idiopathic Basal Ganglia Calcification
Xiaohua Dai,1# Yong Gao,1# Zhenping Xu,1,2# Xiaoniu Cui,1 Juan Liu,1 Yulei Li,1,2
Haibo Xu,3 Mugen Liu,1 Qing K Wang,1,4,* and Jing Yu Liu 1,*
1Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life
Science and Technology, Center for Human Genome Research, Huazhong University
of Science and Technology, 430074, Wuan, Hubei, P. R. China;2Department of Life
Science and Technique, Xinxiang Medical University, Xinxiang, 453003,
China;3Department of Radiology, Union Hospital of Huazhong University of Science
and Technology, Wuhan, 430074, China;4Department of Molecular Cardiology, Lerner
Research Institute, Cleveland Clinic, Cleveland 44195, USA
#Contributed equally to this work.
Objectives:Idiopathic basal ganglia calcification (IBGC) is a neurodegenerative
disorder that is characterized by basal ganglia and extrabasal ganglia calcification, and
usually inherited in an autosomal dominant pattern. To date, two genetic loci for IBGC
were identified on chromosomes 14q and 2q, but further genetic heterogeneity clearly
exists. Methods: Linkage analysis was used to identify the chromosomal location of the
disease gene.Results: A large five-generation Chinese family from Henan Provience
with an autosomal dominant inheritance, family IBGC have identified and characterized.
Linkage analysis excluded the 14q13 and 2q37 loci and other known related genes with
extrapyramidal disease in the nervous system, including ApoE, Tau, ACT, BChE-K,
APP, PS1, PS2 and VLDL-R. Using the family, significant linkage was identified with
markers on chromosome 8p21.1-q11.23 with a maximum LOD score of 4.10. Fine
mapping defined the new genetic locus within a 25.0 cM region between markers
D8S1809 and D8S1833. Interpretation: Future studies of the candidate genes at the
8p21.1-q11.23 locus may lead to identification of a disease-causing gene with IBGC.
Key Words: Idiopathic basal ganglia calcification (IBGC); Linkage analysis; Genetics;
Calcification; LOD score
*Correspondence to:
Dr. J. Y. Liu, liujy@mail.hust.edu.cn
Dr. Q. K. Wang, qkwng@mail.hust.edu.cn

Phosphoinositide-3-Kinase Pathway Activation in PTEN Deficient
Prostate Cancer Cells is Independent of Receptor Tyrosine Kinases and
Mediated by the p110 or p110 Catalytic Subunits
Xinnong Jiang1,2, Sen Chen1, John Asara1, Steven P. Balk1,*
1Division of Hematology-Oncology, Beth Israel Deaconess Medical Center/Harvard
Medical School, Boston, MA, USA
2 Huazhong University of Science and Technology, College of Life Science and
Technology 1037 Luoyu Road, Wuhan, Hubei
Class IA phosphoinositide-3-kinase (PI3K) p110 catalytic subunits are activated upon
SH2 domain mediated binding of their p85 regulatory subunits to tyrosine
phosphorylated pYxxM motifs in receptor tyrosine kinases (RTKs) or adaptor proteins.
The PI3K pathway is activated by PTEN loss in the majority of prostate cancers (PCa),
but the contribution of upstream RTKs that may be targeted therapeutically has not been
assessed. Immunoblotting of p85 associated proteins in serum starved PTEN deficient
LNCaP and C4-2 PCa cells showed a small set of discrete tyrosine phosphorylated
proteins, but these proteins were not recognized by an anti-pYxxM motif antibody and
were not observed in PTEN deficient PC3 PCa cells. LC/MS/MS and immunoblotting
showed that p85 was associated primarily with p110b and p110d. An interaction with
ErbB3 was also detected in serum starved cells LNCaP cells, but was independent of
ErbB3 tyrosine phosphoryation and was not required for basal PI3K activity. Basal
tyrosine phosphorylation of p110 and p110 could be blcoked by c-Src inhibitors,
but this did not suppress basal PI3K activity, which was similarly independent of Ras.
Using siRNA specific for each p110 isoform we found that basal PI3K activity was
mediated by p110 in PC3 cells, and by both p110 �and p110 �in LNCaP cells, while
p110 was required for PI3K activation in response to RTK stimulation by
heregulin- 1. Taken together, these findings indicate that basal PI3K activity in PTEN
deficient PCa cells is RTK independent and mediated by p110 and p110.

Insulin-Like Signaling Pathway Functions in Integrative Response to an
Olfactory and a Gustatory Stimuli in Caenorhabditis elegans
Ya-Ming Jiu1, Zheng-Xing Wu1 , Tao Xu 1,2,
1Key Laboratory of Molecular Biophysics, Ministry of Education, and Institute of
Biophysics and Biochemistry, Huazhong University of Science and Technology, Luoyu
Road 1037#, Wuhan 430074, PR China
2 National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese
Academy of Sciences, Beijing 100101, PR China
Animals integrate various environmental stimuli within the nervous system to generate
proper behavioral responses. However, the underlying neural circuits and molecular
mechanisms are largely unknown. The insulin-like signaling pathway is known to
regulate dauer formation, fat metabolism, and longevity in Caenorhabditis elegans (C.
elegans). Here, we show that this highly conserved signaling pathway also functions in
the integrative response to an olfactory diacetyl and a gustatory Cu 2+ stimuli. Worms of
wild-type N2 Bristol displayed a strong avoidance to the Cu 2+ barrier in the migration
pathway to the attractive diacetyl. Mutants of daf-2 (insulin receptor), daf-18 (PTEN
lipid phosphatase), pdk-1 (phosphoinositide-dependent kinase), akt-1/-2 (Akt/PKB
kinase) and sgk-1 (serum- and glucocorticoid-inducible kinase) show severe defects in
the elusion from the Cu 2+ . Mutations in DAF-16, a forkhead-type transcriptional factor,
suppress the integrative defects of daf-2 and akt-1/-2 mutants. We further report that
neither cGMP nor TGFβ pathways, two other dauer formation regulators, likely plays a
role in the integrative learning. These results suggest that the insulin-like signaling
pathway constitutes an essential component for sensory integration and decision-making
behavior plasticity.
Key Words: C. elegans, insulin signaling pathway, sensory integration, decision
making behavior
Author: Ya-Ming Jiu, Email: hustjym@163.com, Tel: 15110039473

Akt Overexpression Opposes the Beta Amyloid Toxic Injuries to
HEK293/tau Cells
Yin Gang,Zhou xin-wen,Wang jian-zhi
Department of pathology and pathophysiology, the key laboratory of neurological
diseases of the ministry of education ,tongji medical college,huazhong university of
science and technology,wuhan 430030,china
Aim:To study the effect of a direct akt overexpression on Aβ-induced toxic injuries to
neuronal cells.. Methods:To evaluate the effect of Aβ-induced toxic injuries on
neuronal cells,we treated HEK293/tau neurons with several doses of Aβ for 24h.cells
were simultaneously transfected with several doses of a gene akt for 48h. CCK-8 KIT
assay the cell viability,and Reactive oxygen species assay kit assay the level of the level
of the Reactive oxygen species.the apoptosis was detected by flow cytometry.Results:
CCK-8 KIT assay showed that Aβ decreased neuronal cell viability in a
concentration-dependent manner and also that akt overexpression effectively prevented
Aβ-induced neuronal cell death. Aβ treatments increased ROS production in
HEK293/tau cells. Overexpression of Akt but not Akt mutant in the HEK293/tau cells
significantly attenuated Aβ induced ROS production and lipids peroxidation.
Furthermore, overexpression of Akt also attenuated Aβ-induced mitochondrial
dysfunction and apoptosis, and promoted cell survival in HEK293/tau cells.Conclusion:
the neurotoxic effect of Aβ can be partially prevented by akt overexpression.
Key Words: amyloid-beta;akt overexpression; toxic injury;oxidative stress
Author: Yin gang (1977-) was born in xiantao hubei,now I am study in Huazhong
University of Science and Technology for Ph.D, The direction of reseach is the
diagnosis and treatment of alzheimer’s disease. Email: 7706190@163.com, Tel:
13986672302

NMDA Receptors Subunit NR2B Messenger Rnas Expression with
Cognitive Dysfunction after Bilateral Common Carotid Artery Occlusion
in the Adult Rats
Lianjun Guo, Yang Cheng, Bo Zhao
Department of Pharmacology, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan 430030, P.R. China
Aim: To explore whether and how NR2B mRNA may be altered in global incomplete
chronic cerebral ischemic rat model. Methods: Chronic cerebral hypoperfusion in Male
SD rats was performed by permanent occlusion of the bilateral common carotid artery
(2VO) for 30 days. Morris water maze was used to measure spatial learning and
memory performance. Electrophysiological equipments were used to record long-term
potentiation (LTP). In situ hybridization and reverse transcriptase polymerase chain
reaction (RT-PCR) assays were used to investigate NR2B mRNA may be altered in
global incomplete chronic cerebral ischemic rat model. Results: the spatial learning
and memory function of rats attenuated by longer escape latency, shorter time spent in
the target quadrant and impaired long term potentiation (LTP) after chronic cerebral
ischemia. In the in situ hybridization experiment, NR2B declined to 58.82% and
45.60% of the control values in the neocortex and hippocampal region separately after
chronic cerebral ischemia. NR2B mRNA in the hippocampal region and neocortex was
markedly down regulated by ischemia, reaching 43.57% and 30. 05% of the control
values respectively in the semi-quantitative RT-PCR experiment. Conclusions: Our data
suggest that NR2B subunit plays an important role for the spatial learning and memory
function of rats.
Key Words: NR2B messenger RNA; hippocampal neurons; cerebral ischemia
Acknowledgments: This work was supported by grants from the National Natural
Science Foundation of China (NSFC, No. 30772559) to Lianjun Guo
Author: Lianjun Guo , Tel:02783691763, Email: gljyl@yahoo.com.cn

Effects of ZD7288 on Long-Term Potentiation at Perforant Pathway(PP)
Fibers - CA3 Region Synapse Pathway in Rat Hippocampus in Vivo
Xulin Xu1, Min Zheng1, 2 , Lian-Jun Guo1*,
1Department of Pharmacology, Tongji Medical College, Huazhong University of
Science & Technology, 13 Hongkong Road, Wuhan 430030, China.
2 Department of Pharmacology,Xianning College, 88 Xianning Thoroughfare,
Xianning 437100, China.
Aim: To investigate effects of a hyperpolarization-activated cyclic nucleotide-gated cation
(HCN) channel specific blocker ZD7288 to LTP at perforant pathway (PP) fibers-CA3 region
synapse pathway in rat hippocampus. Methods: The field excitatory postsynaptic potentials
(fEPSPs) of PP—CA3 region synapse in rats was recorded by using the extracellular
recording techniques in vivo. Tetanic stimulation (TS; consisted of 50 trains at 0.5 Hz each
composed of 4 pulses at 500Hz) was administrated to observe the induction and maintenance of
LTP. We observed the effects of ZD7288 on the induction and sustain of LTP in PP—CA3
pathway. Simultaneously, the synapse ultrastructural at hippocampal CA3 region was observed
by transmission electron microscope. Results: ①The fEPSPs amplitude persistently stabilized
at 281.8 ± 6.6% of baseline in 90min after giving tetanic stimulation of PP fibers in
LTP-induction group and significantly higher than that in control group(97.4 ±1.8%)(P < 0.01).
② Local administration of ZD7288 100nmol/L before TS, the induction of LTP was obviously
suppressed. The relative fEPSPs amplitudes in each time point were all significantly lower than
that in LTP-induction group (n=10, P 0.05).③Local administration of ZD7288
100nmol/L at 30min after TS, the fEPSPs amplitudes at each time point in 1h after
administration were all significantly lower than LTP-induction group (n=10, P < 0.01), and the
average values were 95.8 ± 6.4% near to control group(P > 0.05). ④ In the LTP induction group,
the cell ultramicrostructure at hippocampus CA3 region showed that there were many synaptic
vesicles in presynaptic endochylema and many synaptic vesicles like thyrsiform occurred in the
presynaptic end in this synapses. ⑤In the ZD7288 -TS group, the cell ultramicrostructure at
hippocampus CA3 region showed that there were no synaptic glomerulus and the synaptic
vesicles were decreased compared with the LTP induction group. Conclusions: It is suggest
that ZD7288 could produce inhibitive effects on the induction and maintenance of long-term
potentiation at PP-CA3 synapse pathway and this action may be associated with altering the
release of neurotransmitter.
Key Words: ZD7288; HCN channel; PP—CA3 pathway ; LTP
Acknowledgments: This work was supported by grants from the National Natural Science
Foundation of China (NSFC, No. 30772559) to Lianjun Guo
Author: Xulin Xu , Email: xulinxu@yahoo.com , Tel: 027-83691762;
* Correspondence author: Tel: +86 27 83691762, E-mail: gljyl@yahoo.com.cn

Effects of Cscl on Synaptic Transmission at Perforant Pathway (PP)
Fibers - CA3 Region of Hippocampus in Rat
Min Zheng1, 2 ,Xulin Xu1, Lian-Jun Guo1*,
1Department of Pharmacology, Tongji Medical College, Huazhong University of Science &
Technology, 13 Hongkong Road, Wuhan 430030, China.
2 Department of Pharmacology,Xianning College, 88 Xianning Thoroughfare, Xianning
437100, China.
Aim: To investigate the contribution of a HCN channel nonspecific blocker CsCl to synaptic
transmission at PP-CA3 pathway in rat hippocampus. Methods: The population spike (PS) of
PP-CA3 region in rats was recorded by using the extracellular recording techniques in vivo.
Tetanic stimulation(TS; consisted of 50 trains at 0.5 Hz each composed of 4 pulses at 500Hz)
was administrated to observe the induction and maintenance of long-term potentiation(LTP). We
observed the effects of CsCl on the induction and sustain of LTP in PP-CA3 pathway.
Simultaneously, the synapse ultrastructural at hippocampal CA3 region was observed by
transmission electron microscope. Results: ①The PS amplitude level persistently stabilized at
281.8 ± 6.6% of baseline in 90min after giving tetanic stimulation of PP fibers in LTP-induction
group and significantly higher than that in control group(97.4 ±1.8%)(n=10, P < 0.01); ②
Local administration of CsCl 5µmol /L before TS, the relative PS amplitudes in each time point
were all significantly lower than that in LTP-induction group (n=10, P

Embryonic Heart Growth is Regulated by Functional Antagonism
Between GRIDLOCK and GATA5 in Zebrafish
Haibo Jia, Sameer Chopra, Mingwei Ni, xiaoling Jiang, Xiaoqun Guan, Terri T. Ni, Sam
Wells and Tao P. Zhong
Departments of Medicine and Cell & Developmental Biology, Vanderbilt Medical
School, Nashville, TN 37232
Embryonic heart growth is revealed primarily by the proliferation of cardiomyocytes
and an increase in their cell size, but the mechanisms that control heart growth during
development are not well understood. In our current study we present evidence that
the Hairy-related bHLH transcription factor Gridlock (Grl) negatively regulates heart
growth in the zebrafish embryo. We show that a reduction in Grl activity by mutation
causes an increase in myocardial gene expression and cardiomyocyte number, resulting
in the development of hyperplastic hearts. Conversely, elevation of Grl activity causes
differentiating cardiomyocytes to fail to divide and grow, thereby reducing the heart size
in the embryo. These Grl-dependent cardiac growth effects are counterbalanced by
Gata5, a transcriptional activator that promotes cardiomyocyte expansion. The
antagonistic relationship between Grl and Gata5 is mediated through protein
interactions that squelch the Gata5 transcriptional activity to inhibit myocardial gene
expression. Hence, the opposing effects of Grl and Gata5 serve to regulate embryonic
heart growth during development

Quantitative Phosphoproteome Analysis Reveals Signaling Events
Involved in Aroma-Stimulated Mouse Olfactory Bulbs
Lei-Ke Zhang1, Fei Liu2, Xiao-Ping Rao2, Fu-Qiang Xu2, Lin Guo1
1College of Life Sciences, Wuhan University, Wuhan 430072, P. R. China
2 State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics,
Wuhan Centre for Magnetic Resonance, Wuhan Institute of Physics and Mathematics,
Chinese Academy of Sciences,Wuhan 430071, P.R.China
Stimulation of odorant molecules is transduced by olfactory receptor neurons (ORNs)
located in the olfactory epithelium of the nasal cavities. Each ORN projects a single
axon to the main olfactory bulb (MOB), where it terminates on the second-order
neurons of the MOB in glomeruli, the initial site for the coding and processing of odor
information. In the glomeruli, ORN axons terminate on the dendrites of mitral/tufted
cells, which in turn project into higher cortical structures. The activity in the MOB is
determined by the input from the SONs, the centrifugal inputs that include regulatory
and feedback fibers from many brain regions, and the nerurocircuits within the MOB.
Finding the proteins that are involved in responding to olfactory stimulation can help to
reveal the mechanism of sensory information processing. Here, signaling events in
odor-stimulated MOBs were explored using quantitative phosphoproteomic analysis.
Our preliminary results showed that a 10 mg protein mixture from the MOB lysate
resulted in the identification and quantification of >1000 unique phosphorylation sites
and ~700 unique phosphoproteins. Further analysis revealed that many up- or
down-regulated phosphorylation events were known having functions in signal
transductuon after odor stimulating. For example, Thr-34 of DCLK1 isoform 1
(accession number: IPI00468380) was up-regulated 8 fold, and Ser-181 of Pgrmc1
(accession number: IPI00319973) was down-regulated 2.13 fold.

Quantitative Phosphoproteomics Analysis of
GABAB Receptor Signaling
Meng FU1, Chanjuan XU2, Jianfeng LIU2,, Lin GUO1,
1College of Life Science, Wuhan University, Wuhan, Hubei, P. R. China, 430072
2College of Life Science and Technology, Huazhong University of Science and
Technology, Wuhan, Hubei, P. R. China, 430074
The G-protein-coupled receptors (GPCRs) play important role in cell-cell interaction.
GABAB receptor, one of the best known GPCR family members, is activated by the
neurotransmitter GABA and is expressed in most neurons. In the current study,
primary cells from cerebellar granule neurons (CGNs) and cerebra cortex were cultured
using SILAC (stable isotope labeling by amino acid in cell culture) method, with
‘SILAC-Light’ as the control group, and ‘SILAC-Heavy’ treated with baclofen, an
agonist specific to mammalian GABAB receptors. After treatment, cell lysate were
mixed 1:1 (protein ratio), and phosphopeptides were enriched by immobilized metal
affinity chromatography (IMAC) method. A hydrophilic interaction chromatography
(HILIC) fractionation step was used before phosphopeptide sequencing and
quantification by liquid chromatography tandem mass spectrometry (LC-MSMS)
analysis. In the preliminary data, we obtained quantitative information for 727
phosphopeptides from cerebra cortex and 254 phosphopeptides from CGNs.
Key wors: Phosphoproteomics, GABAB Receptor, Cell signaling,

Autotaxin Signaling via Lysophosphatidic Acid Receptors Contributes to
LPC-Induced Vascular Smooth Muscle Cell Proliferation and Migration
Bao liang, Zhao Gui-jun, Zhang qian,Yang wen-hua, Yang De-zhi, Li Ying-hui,Alatan
Gaole*
(Department of Biochestry and Molecular Biology ,College ofLife Sciences,Inner
Mongolia University,Hohhot,010021)
Background: Lysophosphatidylcholine (LPC) is a major bioactive phospholipid
component of oxidized low-density lipoprotein (Ox-LDL) and is implicated as a critical
factor in the atherogenic activity of Ox-LDL. LPC is believed to play an important role
in atherosclerosis and inflammatory diseases by altering various functions in a number
of cell-types. We have examined that LPC stimulated the proliferation and migratory
responses of cultured human vascular smooth muscle cells(VSMC),and these actions
were inhibited by Ki16425,a receptor antagonist for the ATX enzymatic product
lysophosphatidic acid (LPA) . We also find that ATX were strongly expressed in the
cells. However, the ATX product mediating the increase in cellular proliferation and
motility remain to be identified. Here,we reported that ATX expressed in the cells
contributes to Lpc-induced VSMC Proliferation and migration .Results: LPC and LPA
stimulated hVSMCs proliferation and migration in a PTX-sensitive manner. These
actions were suppressed by using SiRNA sepecific to ATX and LPA1 receptor,Ki16425.
Furthermore, The LPC- and LPA-induced proliferation of hVSMCs was markedly
inhibited by a potent calcineurin inhibitor cyclosporine A and PD98059,a ERK inhibitor.
Instead, an epidermal growth factor (EGF) receptor tyrosine kinase inhibitor and P38
inhibitor markedly suppressed the migration response to LPC and LPA without having
any significant effect on proliferation of VSMC. .Conclusion: The LPC-induced
stimulation of proliferation in VSMC is mediated by the ATX enzymatic product LPA
through LPA1receptors / Gi/o-proteins/ ERK- MAP kinase /Ca 2+ /calcineurin pathways .
The migration response to LPC and LPA are mediated by the ATX enzymatic product
LPA through LPA1-receptors / Gi/o-proteins/ P38- MAP kinase signaling pathway and
also mediated by transactivation of EGF receptor pathways .
Key words: Autotaxin, Lysophosphatidylcholine, Vascular smooth muscle
cells(VSMC), LPA-receptors signaling pathway
Acknowledge: Supported by the National Science Foundation of China (Grant No.
30760052), was also supported in part by Fostering Talents in Basic Research of the
National Natural Science Foundation of China (Grant No.J0730648 ).
Author*: Alatan gaole, PhD, Professer , Department of Biochemstry and Molecular
Biology , College of Life Sciences , Inner Mongolia University. Adress:No.235,West
University Street, Hohhot, Inner Mongolia, 010021,Tel:0471-4993197, E-mail:
bigaole@imu.edu.cn

Morphine Regulated Thioredoxin-1 Expression
in Neuroblastoma SH-SY5Y Cells
Yuemei Feng, Kui Li, Shengdong Wang, Yanhui Li, Junying Song, Fucheng Luo, Jie
Bai*
College of life science and technology, Kunming University of Science and Technology,
Kunming 650224, China
Aim: Thioredoxin-1 (TRX-1) plays multiple roles in regulation of redox, inhibition of
apoptosis and regulation of transcription factors. Recent studies showed that morphine
could cause oxidative stress in cells and tissues. The aim of current study is to
investigate the expression of TRX-1 under morphine administration in human
neuroblastoma SH-SY5Y cells. Methods: Cell culture, Western blotting. Results: The
expression of TRX-1 was decreased after 1-2h then increased from 4 -24h after
morphine treatment in human neuroblastoma SH-SY5Y. The decreased expression of
TRX-1 by morphine was restored by an inhibitor of GSK3β, lithium chloride. The
increased expression of TRX-1 induced by morphine was suppressed by LY294002,
which is one of the PI3K inhibitor and was suppressed by the antagonist of opioid
receptor, naloxone. The increased expression of TRX-1 recovered to normal level after
morphine withdrawal. These data suggested that TRX-1 was induced through the opioid
receptor and the PI3K/GSK3β pathway. The expression of TRX-1 is dependent on the
morphine administration. Conclusion: Our study suggested that TRX-1 might be
associated with morphine tolerance and abuse.
Key words: thioredoxin-1, morphine, neuroblastoma SH-SY5Y cells
*Author for correspondence: Tel.: +86 13354980126; fax: +86 8713801191, E-mail:
jiebai662001@yahoo.com.cn

Cyclin D1 Promotes Anchorage-independent Cell Survival by Inhibiting
FOXO-Mediated Anoikis
Lu Gan 1, Xiangliang Yang 1, Huibi Xu 1, Haojie Huang 2
1. College of Life Science and Technology, Huazhong University of Science and
Technology, Wuhan 430074, China
2. Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA
Aim: Adhesion to an appropriate extracellular matrix (ECM) is important for normal
cells to survive, and detachment from such supportive matrices usually triggers a
specific type of apoptosis termed anoikis. However, most cancer cells are commonly
resistant to anoikis and can survive in the presence of an inappropriate ECM or in the
absence of anchorage to the ECM. O-class Forkhead box (FOXO) transcription factors
are critical regulators of diverse cellular processes, including apoptosis, cell-cycle arrest,
DNA damage repair and oxidative stress resistance. In this study we determined
whether FOXOs are involved in anoikis and its mechanism. Methods: Annexin V-PE
apoptosis detection kit and soft agar colony formation assay were used to measure the
anoikis in prostate cancer cells. FOXOs activities were determined using a generic
FOXO luciferase reporter gene that contains three copies of forkhead response elements.
Immunoprecipitation was used to determine the endogenous and exogenous interaction
between cyclin D1 and FOXO. In vitro protein binding assay was used to confirm the
interaction between the two proteins. In vitro kinase activity determined whether
FOXO1 was phosphorylated by CDK4/CDK6. Results: FOXO1 and FOXO3a play an
essential role in promoting cell detachment-induced anoikis. In contrast, elevated cyclin
D1 inhibits anoikis. We further show that cyclin D1 interacts with FOXO proteins and
impedes their transcriptional regulatory and anoikis-promoting functions. This effect of
cyclin D1 requires its transcription repression domain but is independent of
cyclin-dependent kinases CDK4 and CDK6. Moreover, we demonstrate that
cancer-derived mutants of cyclin D1 are much more stable than wild-type cyclin D1
under anchorage-independent conditions and possess a greater antagonistic effect on
FOXO-regulated anoikis and anchorage-independent growth of cancer cells.
Conculsion: These data suggest that cyclin D1 may play a critical role in tumorigenesis
and cancer metastasis by inhibiting the anoikis-promoting function of FOXO proteins.
Keywords: Foxo; cyclin D1; Anoikis; protein-protein interaction
Author: Lu Gan,PhD, College of Life Science and Technology, Huazhong University
of Science and Technology, Wuhan 430074, China, Tel: 86-27-87792147
E-mail: lugan@mail.hust.edu.cn

Investigate the Role of Rab27 in the Dcvs Docking and Priming Process
in C. elegans
Wan-juan Feng
Key Laboratory of Molecular Biophysics, Ministry of Education, College of Life Science
& Technology, Huazhong University of Science & Technology
Objective Rab27 is a member of a large family of Ras-related small GTPases. Recent
studies have suggested that Rab27 play an important role in dense-core vesicle
exocytosis. However, which step does Rab27 take part in and what is the precise
mechanisms by which the GTPases regulate dense-core vesicle exocytosis are not clear.
Methods We have succeeded in monitoring the docking and fusion of single dense core
vesicles (DCVs) employing total internal reflection fluorescence microscopy (TIRFM)
in primary cultured C. elegans neurons. Results Using these approaches, we provide
direct evidence that Rab27 regulates the docking step of DCVs at the plasma
membrane in C. elegans.
Keywords: Rab27, total internal reflection fluorescence microscopy(TIRFM), dense
core vesicle(DCV)
Author: E-mail: wanjuanfeng@gmail.com

Biophotons as Neural Communication Signals
Yan Sun, Chao-Wang, Jia-pei Dai *
Wuhan Institute for Neuroscience and Neuroengineering, South-Central
University for Nationalities, Wuhan 430074, China
Aim: Cell to cell communication by biophotons has been demonstrated in plants,
bacteria and certain animal cells. Whether such a signal communication way is existed
in neural cells is unclear. Methods: Here by developing a new method, named as in situ
biophoton autography, we investigated the biophotonic activity in rat spinal nerve roots
in vitro. Results: We found that different spectral light stimulations (infrared, red,
yellow, blue, green and white) can generate biophotons that conduct along the neural
fibers. Such an effect can be significantly inhibited by procaine (a neural conduction
blocker) or classic metabolic inhibitors, suggesting that biophotons may serve as neural
communication signals. The mechanism of biophotonic conduction along neural fibers
may be mediated via protein-protein biophotonic interactions. Conclusion: Biophtonic
activities might be an important way for neural information exchange (biophotonic
transmission) besides well-demonstrated electrical and chemical transmission. This
study may provide a new thought for better understanding of the fundamental
mechanisms of neural communications and the functions of nervous system such as
vision, learning and memory, as well as the mechanisms in human neurological
diseases.
Keywords: Biophotons, in situ biophoton autography, neural communication signals,
spinal nerve root.
* Corresponding author
E-mail: jdai@mail.scuec.edu.cn; Tel: 0086-27-67840917

Effects of Cyclic Nucleotide Gated Channel Knockout on the Olfactory
Bulb Analyzed by Quantitative Phospho-Proteomics
Fei Liu 1 , Xiao-Ping Rao, Lei-Ke Zhang 2 , Lin Guo 2 , Fuqiang Xu 1
1State Key Laboratory of Magnetic Resonance and Atomic and Molecular
Physics, Wuhan Centre for Magnetic Resonance, Wuhan Institute of Physics
and Mathematics Chinese Academy of Sciences, Wuhan 430071, P. R. China
2College of Life Sciences, Wuhan University, Wuhan 430072, P. R. China
When odorant molecules enter nasal cavity and are dissolved in the mucus covering the
olfactory epithelium, they bind with odorant receptor proteins in the cilia of the
olfactory sensory neurons, leading to a series of signal transduction processes:
activation of trimeric G-proteins and then nucleotide cyclase, to increase cAMP
concentration, opening of cyclic nucleotide gated (CNG) channel, and at last, generating
action potentials that are sent to the olfactory bulb. CNG channel, mainly expressed in
visual and olfactory systems, is non-selective to cations. It is necessary to lead to
depolarization of the olfactory sensory neurons. Knockout of one of its subunit, cnga2,
give rise to anosmic mice. The transgenic animals have much lower surviving rate,
thinner olfactory epithelium, smaller olfactory bylb and less aggressive behavior. To
investigate the effects of long term absence of peripheral stimulation on the olfactory
bulb, we used quantitative phosphor-proteomics to analyze the change of protein
phosphorylation. We found hundreds of phosphorylated proteins and even more
phosphorylated sites. Generally, proteins with significant changes in phosphorylation
are these related to signal transduction, neuronal transmission, homeostasis and
neurogenesis. For examples, GAD1, a GABA synthase, is decrease to 19%; CAMK2b, a
kinase, to 33%; and Slc4a4, a electrogenic sodium bicarbonate cotransporter, to 17%
of the control, while Hsp90aa1, heat shock protein HSP 90-alpha, increased by a
101%; and Alpl, a tissue-nonspecific alkaline phosphatase by 346%. The exact roles of
these proteins in the olfactory bulb are under investigation.
Key words: olfactory bulbs, transgenic mice, phosphoproteins, IMAC/LC-MS