conference booklet - MDC

International Conference „Development of Somatosensation and Pain 2008“ 

Welcome Address 

Dear Friends and Colleagues, 

It's our pleasure to invite you to the International Berlin Spring Meeting 2008 

"Development and function of somatosensation and pain" in the Max Delbrück 

Communications Center in Berlin-Buch from 14 - 17 May 2008. 

The aim of the meeting would be to bring together researchers interested in the 

development and function of the somatosensory system. Much progress has been 

made in recent years in elucidating the molecular mechanisms responsible for the 

phenotypic differentiation of primary afferent neurons as well as spinal dorsal horn 

neuronal circuits to which sensory neurons connect. We now also understand a lot 

more about the molecules (ion channels and other proteins) that allow sensory 

neurons to detect biologically relevant thermal and mechanical stimuli. 

The aim of this conference would be draw together top researchers working in these 

two related fields. This unique constellation of researchers would allow participants to 

explore the idea that the development of the somatosensory system may in turn help 

us also to understand its functional organization and pathology in human disease. 

The Max Delbrück Center (MDC) Conferences are international symposia dedicated to 

timely topics in Molecular Medicine and Translational Research. The Conference 

Series aims at fostering exchange of scientific information in all areas relevant to 

modern biomedical sciences including cell and developmental biology; molecular 

cardiovascular, neuroscience and oncology research; genetics, genomics and 

bioinformatics; as well as molecular pharmacology. 

You are invited to participate in what promises to become an exciting scientific 

meeting. We look forward to welcoming you in Berlin, an exiting European capital city. 

Yours sincerely, 

The Organizer 

Gary Lewin 

1 Welcome Address

Organizers & Scientific Advisory Board 

Organizers 

• Gary Lewin – Berlin, Germany 

• Carmen Birchmeier – Berlin, Germany 

• Ines Ibanez Talon – Berlin, Germany 

• Michael Schaefer – Berlin, Germany 

• Christoph Stein – Berlin, Germany 

Scientific Advisory Board 

• Carlos Belmonte – Alicante, Spain 

• Patrik Ernfors – Stockholm, Sweden 

• Martyn Goulding – La Jolla, USA 

• Thomas Jessell – New York, USA 

• Qiufu Ma – Boston/Cambridge, USA 

May 14 – May 17, 2008 – Berlin, Germany 

Organizers & Scientific Advisory Board 2


Organization 

Michaela Langer / Jana Droese 

Max Delbrück Center for Molecular Medicine (MDC) Berlin-Buch 

Robert-Rössle-Str. 10 

13125 Berlin 

phone: +49 30 9406 3720 / 4254 

fax: +49 30 9406 2206 / 2170 

email: langer@mdc-berlin.de / jana.droese@mdc-berlin.de 

Conference Office: 

Phone : +49 30 9406 3720 / 4824 

Fax: +49 30 9406 2206 / 2170 

3 Organization

Conference Information 


Venue: 

Max Delbrück Communications Center (MDC.C), Robert-Rössle-Str. 10, 13125 

Berlin-Buch 

Germany 

Contact: 

Conference secretariat (MDC.C), Michaela M. Langer, 

phone: +49 30 9406 3720/4824 / fax: +49 30 9406 2206 / e-mail: langer@mdcberlin.de 

Date: 

From Wednesday, May 14, 2008 afternoon to Saturday, May 17, 2008 3:00 p.m. 

Registration fee (late): 

Full registration fee 470 € 

Trainees 270 € 

Daily registration 100 € 

Conference Dinner 50 € 

The registration fee includes: Access to scientific sessions; Program and abstract 

book; free access to the Internet; Coffee breaks; Working lunches; Welcome reception 

and Evening Buffet. The Conference Dinner is NOT included in the registration fee. 

Posters: Posters will be displayed during the meeting close to the conference hall. 

The size of a single poster should be no more than 1m (horizontally) x 1.20m 

(vertically). 

In this abstract volume, you can find a number for your abstract and according to 

these numbers your poster can be mounted in the exhibition room. 

Mounting material will be available from the registration desk. 

Internet café will be located in the 3rd floor, Seminar room Dendrit 1. WLAN is 

available. 

Social events: 

* Welcome reception 

Wednesday, May 14, 7.15 p.m. – 9.30 p.m. - Venue: Max Delbrück Communications 

Center (MDC.C) 

* Evening Buffet with Poster Session 

Thursday, May 15, 6.00 p.m. – 8.00 p.m. - Venue: Max Delbrück Communications 

Center (MDC.C) 

* Conference Dinner 

Friday, May 16, 7.00 p.m. – downtown – 

Venue: Restaurant “Guy” Am Gendarmenmarkt, Jägerstrasse 59-60, 10117 Berlin 

Information at the Registration desk 

Buses 

Please notice the information at the Registration desk every day! 

Conference Information 4


Table of Contents 

Conference Information 4 

Program 6 - 11 

Speakers Abstracts 12 - 42 

List of Posters 43 - 46 

Poster Abstracts 47 - 74 

List of Participants 75 - 82 

Author Index 83 - 85 

5 Table of Contents

Program 

WEDNESDAY, May 14, Plenary Lectures 

18:00 – 18:10 Introduction 

Gary Lewin 

18:10 – 19:10 Using Pain to Block Pain 

Clifford Woolf 

19:10 Welcome Buffet 


Program 6


THURSDAY, May 15 

7 Program 

Development of Sensory Pathways I 

Chair: Carmen Birchmeier 

09:00 – 09:30 A new mechanoreception assay reveals intrinsic differences 

between sensory subtypes 

Patrik Ernfors 

09:30 – 10:00 Molecular characterization of low-threshold mechanoreceptor subtypes. 

Patrick Carroll 

10:00 – 10:15 The homeodomain factor Lbx1 controls the differentiation of 

sensory relay neurons in the hindbrain 

Robert Storm 

10:15 – 10:45 Coffee Break 

10:45 – 11:00 A cGMP signalling pathway essential for sensory axon bifurcation 

Hannes Schmidt 

11:00 – 11:30 NGF signalling and the control of gene expression in developing 

somatosensory neurons. 

David Ginty 

11:30 – 14:00 LUNCH (Buffet) and Poster session 

Sensory Transduction Mechanisms I 

Chair: Gary Lewin 

14:00 – 14:30 Deconstructing the molecular events responsible for touch and 

temperature sensation in C. elegans 

Miriam Goodman 

14:30 – 14:45 Defining a function for the ion channel TRPA1 

Sandra Zurborg 

14:45 – 15:15 The K2P channels: focus on TREK-1 

Eric Honore 

15:15 – 15.30 SCF/c-Kit is required for normal noxious heat sensitivity 

Alistair Gerratt 

15:30 – 16:00 Modulation of thermo-TRP ion channels by phosphorylation 

Peter McNaughton 

16:00 – 16:30 Coffee Break

Pain Mechanisms I 

Chair: Ines Ibanez Tallon 

16:30 – 17:00 Trk(ing) on from Pain. 

Lorne Mendell 


17:00 – 17:30 Neuronal circuits and receptors involved in spinal cord pain 

processing 

Andrew Todd 

17:30 – 18:00 New insights into mechanisms of spinal LTP and pain memory 

Rohini Kuner 

18:00 – 20:00 Poster Session (with CHEESE and WINE) 

Program 8


FRIDAY, May 16 

09:00 – 09:30 TBA 

Martyn Goulding 

9 Program 

Development of Somotosensory pathways II 

Chair: Alistair Garratt 

09:30 – 10:00 Transcriptional Control in Dorsal Spinal Cord Development: New 

Roles for Old Factors 

Jane Johnson 

10:00 – 10:15 The Novel Estrogen Receptor GPR30 Mediates Estrogen-Induced 

And PKCepsilon -Dependent Mechanical Hyperalgesia In Vitro 

And In Vivo 

Tim Hucho 


10:45 – 11:00 Adaptations in the C Fiber Nociceptors of Naked Mole-Rats 

Render These Animals Insensitive to Specific Air-Borne Irritants 

and CO2-Induced Pulmonary Edema 

Thomas Park 

11:00 – 11:30 Tactile Experience Shapes Behaviour in Etruscan Shrews 

Michael Brecht 

11:30 – 12:00 Specification and differentiation of dorsal spinal cord interneurons. 

Carmen Birchmeier 

12:00 – 14:00 LUNCH (Buffet) and Poster Sessions 

Pain Mechanisms II 

Chair: Christoph Stein 

14:00 – 14:30 The menthol receptor TRPM8 is the principal detector of 

environmental cold 

Jan Siemens 

14:30 – 14:45 Pain attenuation in Tethered-Toxin- transgenic mice due to 

reduced excitability of sensory neurons 

Ines Ibanez-Tallon 

14:45 – 15:15 Acid-Sensing Ion Channels (ASICs) and pain in the central and 

peripheral 

Eric Lingueglia 

15:30 – 16:00 Coffee Break and Poster Session 

15:15 – 15:30 Leukocyte-derived and exogenous opioids acting at peripheral 

opioid receptors control neuropathic pain 

Dominika Labuz


16:00 – 16:30 Synaptic Dis-Inhibition in Pathological Pain States 

Hanns-Ulrich Zeilhofer 

19:00 GET TOGETHER and Congress Dinner (downtown) 

Program 10


SATURDAY, May 17 

11 Program 

Transduction, Development, and Pain 

Chair: Michael Schaefer 

09:00 – 09:30 Mechanosensitive ion channels, stomatin-like proteins and 

molecular tethers essential for touch. 

Gary Lewin 

09:30 – 10:00 Sensory Neuron Mechanotransduction 

John Wood 

10:00 – 10:30 Breathing with Phox2b 

Jean-Francois Brunet 


Transduction, Development, and Pain 

Chair: Robert Schmidt 

11:00 – 11:30 Roles of Runx1 in controlling nociceptor development and pain 

behaviors 

Qiufu Ma 

11:30 – 12:00 Peripheral mechanisms for cold detection in intact and injured 

tissues 

Carlos Belmonte 

12:00 – 12:30 Genetic Determinants of Pain 

Stephen McMahon 

12:30 – 14:00 LUNCH (Buffet)

Speaker Abstracts 


Speaker Abstracts 12


T1 Using Pain to Bock Pain 

Clifford J. Woolf 

Department of Anesthesia and Critical Care, Massachusetts General Hospital and 

Harvard Medical School, Boston USA 

We find that it is possible to selectively block electrical signaling in nociceptors without 

affecting signaling by other neurons (Binshtok et al Nature 449: 607-611, 2007). The 

method is based on introducing permanently charged sodium channel blockers 

through the pore of TRPV1 channels which are present on nociceptors but not motor, 

autonomic, or low threshold mechanosensitive neurons. TRPV1 channels are 

activated by noxious heat and protons, and also by pungent ligands such as 

capsaicin. The pore of the TRPV1 channel is large enough to pass N-methyl-lidocaine 

(QX-314), a positively charged hydrophilic lidocaine derivative that is normally 

ineffective as a local anesthetic because it cannot permeate through the membrane to 

access the lidocaine-binding site on the inner face of voltage-gated sodium channels. 

By co-applying capsaicin and N-methyl-lidocaine, it is possible to introduce the Nmethyl-lidocaine 

selectively into nociceptors, where it blocksTTX-sensitive and 

resistant sodium channels in these neurons to prevent action potential firing. Unlike 

conventional local anesthetics, which block electrical activity in all neurons, this 

produces a local anesthesia without paralysis or numbness, effectively creating a local 

analgesia. Also, the inhibition of pain lasts longer than conventional local anesthetics, 

since the charged sodium channel blocker is effectively trapped inside the nociceptor 

after the two agents are applied and removed. This approach should lead to the 

development of novel combination analgesics comprising agonists of TRP channels to 

activate intrinsic drug delivery systems, and impermeant cationic ion channel blockers 

whose acces to select cell types is determined by the expression profile of the TRPs. 

13 Speaker Abstracts


T2 A new mechanoreception assay reveals intrinsic differences between 

sensory subtypes 


Division of Molecular Neurobiology, Department of Medical Biochemistry and 

Biophysics Scheelesv 1 Karolinska Institute, S-171 77 Stockholm, Sweden 

The somatic sensory system comprises different perceptual modalities. Neurons of the 

dorsal root ganglion mediate tactile sensation by mechanoreceptive stimuli, limb 

proprioceptive sensation elicited by displacement and the static tension of muscles, 

and pain and thermal sensations. .A new functional in vitro assay has been 

developed. Groups of neurons have similar response profiles and different neurons 

respond to different types and levels of mechanical stimuli. Hence, this in vitro assay 

can discriminate between different mechanoreceptors in terms of their stimuli 

response profile. This suggests that inherent differences in vivo is preserved in vitro 

and could thus be a selectable marker for a neuronal subtype. In this assay, nerve 

ending are subjected to pressure in a three dimensional matrix which leads to an 

action potential to the cell body where measurements are taken, and is thus very 

similar to the in vivo situation where mechanosensation is generated in the periphery 

due to local deformation or vibration. 



T3 Molecular characterization of low-threshold mechanoreceptor sub-types. 


INSERM U.583, Physiopathologie et thérapie des déficits sensoriels et moteurs. 

Institut des Neurosciences de Montpellier (INM), Hopital St. Eloi, 80 rue Augustin 

Fliche, BP 74103, 34091 Montpellier cedex 5 

Low-threshold mechanoreceptor neurons of the dorsal root ganglia (DRG) have been 

difficult to study because of the dearth of specific molecular markers of these celltypes. 

In a screen for genes expressed in specific DRG sub-types we found that a 

transcription factor of the large Maf family (Maf-A) is expressed in 8-10% of embryonic 

DRG neurons. By co-localization studies with known markers of sensory neuron subtypes 

MafA was found to be exclusively co-localized with the tyrosine kinase receptor 

c-Ret. Double Maf-A+/Ret+ neurons are observed in the DRG at early developmental 

stages (E11 – E13) before the appearance of c-Ret in nociceptors at late embryonic 

stages. Analysis of the peripheral and central projections of these neurons at P0 

suggests that they are slowly-adapting mechanoreceptors. Preliminary 

characterization, using mutant mice, of the roles MafA and c-Ret in these 

mechanoreceptors will be presented. 



T4 The homeodomain factor Lbx1 controls the differentiation of sensory relay 

neurons in the hindbrain 

Robert Storm, Thomas Müller, Martin Sieber, Carmen Birchmeier 

Max-Delbrück-Centrum für Molekulare Medizin (MDC) 

The homeodomain factor Lbx1 is expressed in postmitotic neurons in the ventral alar 

plate of the developing hindbrain extending from rhombomere 2 into the spinal cord. 

Lbx1 expression distinguishes two major classes of postmitotic neurons: class A 

(Lbx1-negative) and class B (Lbx1-positive). Here, we provide a classification of the 

neuronal subtypes emerging in the alar plate of the hindbrain according to their 

transcription factor profiles. Genetic lineage tracing allowed us to follow Lbx1 

derivatives throughout embryonic development in control and Lbx1 mutant mice. We 

show that most of the Pax2+, GABAergic neurons in the caudal hindbrain and Lmx1b+ 

somatosensory neurons of the spinal trigeminal nucleus are Lbx1 derivatives. In Lbx1 

mutant mice, dorsal Pax2+ neurons are not specified and the spinal trigeminal nucleus 

is not formed. Instead, the viscerosensory nucleus of the solitary tract (NTS) as well 

as the inferior olive (IO) are enlarged. In normal development the NTS and the IO are 

formed by dA3 and dA4 neurons, respectively. We conclude that Lbx1 is a major 

determinant in sensory neuron development, which chooses a somatosensory over a 

viscerosensory and climbing fiber developmental program in the caudal hindbrain. 



T5 A cGMP SIGNALING PATHWAY ESSENTIAL FOR SENSORY AXON 

BIFURCATION 

Hannes Schmidt 1 , Agne Stonkute 1 , René Jüttner 1 , Susanne Schäffer 1 , 

Jens Buttgereit 2 , Robert Feil 3 , Franz Hofmann 3 , Fritz G. Rathjen 1 

1 Max Delbrück Center for Molecular Medicine, Developmental Neurobiology Group, 

Berlin, Germany, 2 Max Delbrück Center for Molecular Medicine, Molecular Biology of 

Peptide Hormones Group, Berlin, Germany, 3 Technical University of Munich, 

Department of Pharmacology and Toxicology, Munich, Germany 

The complex wiring pattern of the mature nervous system is shaped during 

development by the phenomena of axonal guidance and branching. The intracellular 

signal transduction mechanisms underlying these processes are only poorly resolved. 

We identified a signalling axis in sensory neurons essential for one form of axonal 

branching. The central trajectories of dorsal root ganglion axons display at least two 

types of ramifications when they enter the spinal cord: (1) bifurcation at the dorsal root 

entry zone (DREZ) and (2) interstitial branching from stem axons to generate 

collaterals that penetrate the grey matter. We report a cGMP signaling cascade 

critically involved in the establishment of the highly stereotyped pattern of T-shaped 

axon bifurcation of sensory axons at the DREZ of the spinal cord. Single axon labeling 

using DiI revealed that embryonic mice with an inactive receptor guanylyl cyclase 

Npr2 or deficient for cGMP-dependent protein kinase I (cGKI) lack the bifurcation of 

sensory axons at the DREZ, i.e. the ingrowing axon either turns rostrally or caudally 

instead. Cross-breeding experiments of these mutant mice with a mouse line 

expressing EGFP in sensory neurons under control of the Thy-1 promoter 

demonstrate that the bifurcation error is maintained to mature stages. In contrast, 

interstitial branching of collaterals from primary stem axons remains unaffected. At a 

functional level, the distorted axonal branching is accompanied by reduced synaptic 

input, as revealed by patch clamp recordings of neurons in the superficial layers of the 

cord. Hence, our data demonstrate that a cGMP signaling cascade including Npr2 and 

cGKI is essential for axonal bifurcation at the DREZ and influences neuronal 

connectivity in the dorsal spinal cord. 



T6 NGF signaling and the control of gene expression in developing 

somatosensory neurons. 

S. Rasika Wickramasinghe, Rebecca S. Alvania, Narendrakumar Ramanan, 

Kenji Mandai, David D. Ginty 

The Solomon H. Snyder Department of Neuroscience, The Howard Hughes Medical 

Institute, The Johns Hopkins University School of Medicine, 725 N. Wolfe St, PCTB 

1015, Baltimore, MD 21205. 

Nerve Growth Factor (NGF) controls survival, maturation and target innervation by 

cutaneous sensory neurons. We report that Serum Response Factor (SRF), a 

prototypic transcription factor that mediates stimulus-dependent gene expression, is a 

critical mediator of NGF signaling, axonal growth, branching, and target innervation by 

embryonic DRG sensory neurons. Conditional ablation of the murine SRF gene in 

DRGs results in no deficits in sensory neuron viability or differentiation, but causes 

dramatic defects in extension and arborization of peripheral axonal projections in the 

target field in vivo, a phenotype also observed in mice lacking NGF. Moreover, SRF is 

both necessary and sufficient for NGF-dependent axonal outgrowth in vitro, and NGF 

regulates SRF-dependent gene expression and axonal outgrowth through activation of 

both MEK/ERK and MAL signaling pathways. Finally, we found that NGF is essential 

for the expression of several SRF-dependent cytoskeletal genes in embryonic DRG 

neurons in vivo. Together, our findings suggest that SRF is a major effector of both 

MEK/ERK and MAL signaling by NGF, and that SRF is a key mediator of NGFdependent 

target innervation by embryonic sensory neurons. 



T7 Deconstructing the molecular events responsible for touch and temperature 

sensation in C. elegans 


Stanford University, Stanford, CA 94305 USA 

Sensation guides behavior, providing critical information about the environment. The 

senses of touch and vibration are critical for daily activities like standing and walking, 

while temperature sensation is essential for efficient thermoregulation. We seek to 

understand the molecular events that give rise to touch and temperature sensation 

using the nematode Caenorhabditis elegans as a model system. Powerful tools in 

classical and molecular genetics and the ability to record electrical responses to 

sensory stimuli in living animals make this simple roundworm a nearly perfect animal 

for these studies. Our work focuses on the six non-ciliated touch receptor neurons 

(TRNs) that detect touch applied to the body wall and a pair of thermosensory neurons 

(AFD) that detect changes in ambient temperature. Genetic analyses have shown that 

TRN function depends on degenerins encoded by the mec-4 and mec-10 genes, while 

AFD function depends on cGMP-gated ion channels encoded by tax-2 and tax-4. We 

are using in vivo whole-cell patch-clamp recording in concert with genetic dissection to 

investigate the biophysics and molecular basis of touch and temperature transduction. 

We have used this approach to show that MEC-4 and MEC-10 form the pore of native 

mechanotransduction channels and that such channels rely on the MEC-2 and MEC-6 

auxiliary subunits for full functionality in vivo (Nat Neurosci 8: 43). Now, we are 

investigating the mechanism by which these channels are activated in vivo. We have 

also used in vivo whole-cell patch clamp recording to characterize the biophysical and 

molecular basis of temperature-sensitivity in AFD neurons. I will present evidence that 

the AFD neurons detect cooling and warming by modulating the activity of the cGMPgated 

ion channel encoded by tax-4 and tax-2. 


T8 Defining a function for the ion channel TRPA1 


Sandra Zurborg, Brian Yurgionas, Ombretta Caspani, Paul A. Heppenstall 

TRPA1 is a non-selective ion channel belonging to the transient receptor potential 

(TRP) family. The channel is expressed by a subset of pain sensing neurons in the 

peripheral nervous system and can be activated by many pain eliciting compounds 

e.g. acrolein and mustard oil. Its endogenous function remains unclear, although it is 

generally accepted that TRPA1 is important in the pain pathway. In my PhD. project I 

have working on defining a function for TRPA1 at the cellular and whole animal level. 

In initial studies we demonstrated that TRPA1 is directly gated by intracellular Ca2+ 

and can be activated by several stimuli which raise intracellular Ca2+ levels in sensory 

neurons. We identified an EF-hand domain in the N-terminus of TRPA1 and 

demonstrated that it is responsible for the calcium sensitivity of the channel. More 

recently we have been investigating the expression of TRPA1 in mouse models of 

inflammatory pain because changes in the expression of TRPA1 might be a critical 

mechanism for modifying excitability of painsensing neurons after injury. In order to 

model inflammatory pain we injected Complete Freund's Adjuvant (CFA) into the hind 

paw of mice. We assessed TRPA1 expression by means of calcium imaging of 

dissociated dorsal root ganglia (DRG) neurons and in situ hybridization of DRG 

sections after inflammation. We saw no change in the TRPA1 expression when we 

compared untreated mice and inflamed animals. However we did observe significantly 

larger mustard oil evoked responses in CFA-treated mice compared to control mice. 

This suggests that TRPA1 might contribute to enhanced excitability and 

hypersensitivity during inflammation. 



T9 The K2P channels: focus on TREK-1 

Eric Honore 

Institut de Pharmacologie Moléculaire et Cellulaire, CNRS UMR 6097, Université de 

Nice-Sophia Antipolis, 660 route des Lucioles, 06560 Valbonne, France. 

Two-pore-domain K+ (K2P) channel subunits are made up of four transmembrane 

segments and two pore-forming domains that are arranged in tandem and function as 

either homo- or heterodimeric channels. This structural motif is associated with 

unusual gating properties including background channel activity and sensitivity to 

membrane stretch. Moreover, K2P channels are modulated by a variety of cellular 

lipids and pharmacological agents, including polyunsaturated fatty acids and volatile 

general anesthetics. Recent in vivo studies have demonstrated that TREK-1, the most 

thoroughly studied K2P channel, has a key role in the cellular mechanisms of 

neuroprotection, anaesthesia, pain and depression. 


T10 SCF/c-Kit is required for normal noxious heat sensitivity 


Nevena Milenkovic 1 , Christina Frahm Frahm 2 , Carola Griffel 2 , Bettina Erdmann 3 , 

Carmen Birchmeier 2 , Gary R. Lewin 1 , Alistair N. Garratt 2 

1 Molecular Physiology of Somatic Sensation, 2 Developemental biology/Signal 

Transduction, 3 Max-Delbrück Center for Molecular Medicine 

Sensory neurons in the dorsal root ganglion (DRG) transduce diverse sensory 

sensations like touch, heat and pain. C fibers represent about 60% of DRG cell 

population; most of them are polymodal responding both to noxious thermal and 

mechanical stimuli. Neurotrophins are known to be involved in determining the final 

phenotype of sensory neurons. NGF is a main mediator of inflammatory hyperalgesia 

and also critical for the development of normal noxious heat sensitivity. Lower noxious 

heat sensitivity and lower expression level of c-Kit after neonatal NGF deprivation 

make c-Kit a good candidate as mediator of noxious heat sensitivity. The receptor 

tyrosine kinase c-Kit was functionally characterized. In DRG c-Kit is predominately 

expressed in small diameter cells expressing TrkA and CGRP. We showed that 

SCF/c-Kit signaling system has a key role in setting the thermal threshold for 

activation of heat sensing nociceptive neurons. Mice lacking a functional c-Kit receptor 

displayed profound thermal hypoalgesia attributable to a marked elevation in the 

thermal threshold and reduction in spiking rate of CMH nociceptors. Activation of c-Kit 

by SCF resulted in a reduced thermal threshold and profound potentiation of heatactivated 

currents in 50% of isolated heat sensitive small diameter neurons. Acute 

application of SCF induced thermal hyperalgesia in mice and this action required the 

TRP-family cation channel TRPV1. In addition, lack of c-Kit signaling during 

development resulted in hypersensitivity of discrete mechanoreceptive neuronal 

subtypes to mechanical stimulation. The reduced noxious heat sensitivity was 

observed in mice treated with c-Kit blocker, Imatinib. The ability of SCF to potentiate 

capsaicin induced Ca2+ intracellular concentration in isolated DRG neurons is 

currently used to investigate the SCF/c-Kit signaling pathway. Thus c-Kit, can be be 

grouped into a small family of receptor tyrosine kinases, including c- Ret and TrkA, 

that control the transduction properties of distinct types of sensory neuron to thermal 

and mechanical stimuli. 



T11 Modulation of thermo-TRP ion channels by phosphorylation 

Xuming Zhang, Peter McNaughton 

Dept of Pharmacology, University of Cambridge 

The ability of vertebrates to detect and avoid damaging extremes of temperature 

depends on activation of ion channels belonging to the thermo-TRP family. Injury or 

inflammation lowers the threshold for detection of painful levels of heat, a process 

known as heat hyperalgesia. A wide range of inflammatory mediators, amongst which 

the best-studied are bradykinin, prostaglandin E2 and nerve growth factor (NGF), are 

liberated by inflammation or injury and are able to cause heat hyperalgesia. 

Inflammatory mediators activate at least three distinct intracellular signalling pathways 

which lower the threshold of the heat-activated ion channel TRPV1. Bradykinin 

activates PKC-epsilon and phosphorylates two serine residues on TRPV1, while 

prostaglandin E2 activates PKA and phosphorylates a partially overlapping set of 

residues. Both these mechanisms enhance the open probability of TRPV1 channels 

already located in the surface membrane. NGF, on the other hand, activates the 

tyrosine kinase Src to phosphorylate a single tyrosine in the N-terminal domain and so 

to promote movement of new channels from a vesicular store to the surface 

membrane. Some work had suggested that a reducution in membrane PIP2 was a 

major mechanism for modulation of TRPV1, but recent work in our lab and elsewhere 

has now discounted this possibility. 

In recent work we have shown that modulation of the sensitivity of TRPV1 by the 

protein kinases PKA and PKC, and by the phosphatase calcineurin, depends on the 

formation of a signalling complex between the scaffolding protein AKAP79/150 and 

TRPV1. We have identified a critical AKAP79/150 binding region in the TRPV1 Cterminal 

domain. If binding is prevented then sensitisation by both bradykinin and 

PGE2 is completely abrogated. AKAP79/150 is therefore a final common element, 

vital for heat hyperalgesia, on which the effects of multiple pro-inflammatory mediators 

converge. 


T12 Trk(ing) on from Pain 

J. Petruska, V. Boyce, L. M. Mendell 


Department of Neurobiology and Behavior, SUNY- Stony Brook, Stony Brook, NY, 

11794, USA 

Previous work from this laboratory has demonstrated that the neurotrophins NGF and 

BDNF can acutely sensitize peripheral and central components of the nociceptive 

pathway (Mendell, Handbook of the Senses, 2008). Further experiments revealed that 

another neurotrophin, NT-3, acutely sensitizes the monosynaptic spindle- evoked 

EPSP via trkC receptors (Arvanov et al., J. Neurophysiol. 2000). Additional work 

demonstrated that long lasting NT-3 administration to axotomized nerves or to the 

spinal cord during development strengthens synapses made by muscle spindle 

afferent fiber (group Ia) fibers on motoneurons (Mendell et al., J. Neurosci. 1999; 

Arvanian et al., J. Neurosci. 2003). Step training in spinal rats transected as neonates 

improves stepping performance. It also increases the monosynaptic EPSP in ankle 

extensor motoneurons (Petruska et al., J. Neurosci., 2007) which should improve hind 

limb weight bearing and facilitate initiation of the swing phase (“stepping off”). 

Because neurotrophin levels, including NT-3 are elevated in the spinal cord of step- 

trained spinal rats, we have investigated whether direct administration of NT-3 elicits 

effects similar to those of step training in neonatally transected rats. In the present 

work we made use of AAV-NT-3 viral constructs to administer NT-3. When injected 

into peripheral tissues they are transported to the spinal cord and DRG in an AAV 

serotype- dependent manner. NT-3 levels are substantially increased in the spinal 

cord for months after a single administration of the appropriate viral particles into 

hindlimb muscles. We have found electrophysiological changes in treated transected 

rats that are qualitatively similar to those observed after step training. Furthermore, 

these rats show improved stepping ability compared to untreated rats, in general 

agreement with findings in the cat (Boyce et al., J. Neurophysiol., 2007). This work 

may have translational potential. We are using vectors and neurotrophins delivered in 

a manner that is consistent with clinical application. Here we demonstrate that these 

agents elicit useful behavioral effects that may be explained at least in part by the 

electrophysiological changes we measure in the spinal cord. Drs. R. Ichiyama and 

V.R. Edgerton (UCLA) carried out behavioral aspects of this work. Drs. B. Kaspar and 

F. Gage (Salk Institute) designed and prepared the engineered AAV viruses. 

Supported by the Christopher and Dana Reeve Foundation, and NIH (2 RO1 NS 

16996). 



T13 Neuronal circuits and receptors involved in spinal cord pain processing 

Andrew Todd 

Spinal Cord Group, West Medical Building, University of Glasgow, University Avenue, 

Glasgow G12 8QQ, U.K. 

The spinal dorsal horn receives sensory information from primary afferents, and this is 

transmitted to the brain through projection neurons. It also contains numerous 

excitatory and inhibitory interneurons, and receives descending modulatory inputs. 

Despite its importance in pain mechanisms, we still know little about the neuronal 

circuitry within this region. Projection neurons are concentrated in lamina I and 

scattered throughout deeper laminae. Lamina I neurons in lumbar cord project to the 

lateral parabrachial area, periaqueductal grey matter and caudal medulla. There are 

few lamina I spinothalamic neurons in lumbar segments, although these cells are 

common in the cervical enlargement. The targets of lamina I spinothalamic neurons 

include the posterior triangular nucleus, which projects to second somatosensory and 

insular cortices. Most lamina I projection neurons express the neurokinin 1 receptor 

(NK1r). These cells, together with projection neurons in laminae III-IV that also 

express this receptor, are densely innervated by substance P-containing primary 

afferents. This provides a powerful route linking nociceptors with brain regions 

involved in pain. We have also identified a population of large lamina I projection cells 

that lack the NK1 receptor and have a high density of inhibitory synapses. Glutamate 

is the main excitatory transmitter in the dorsal horn and acts on several types of 

receptor, including AMPA receptors (AMPArs), which play a major role in perception of 

somatosensory stimuli. AMPArs are made up from four subunits (GluR1-4), and 

subunit composition determines receptor properties. Synaptic AMPArs are difficult to 

detect with immunocytochemistry because protein cross-linking at synapses following 

fixation. However, they can be revealed with antigen retrieval, and this can be used to 

determine the laminar distribution of different subunits and the subunit composition of 

synaptic AMPArs on different types of neuron. We have found that lamina III/IV NK1rexpressing 

neurons and the large lamina I cells that lack NK1r both have a high 

density of GluR4-containing synapses. Cells in the latter population receive numerous 

synapses from axons that contain vesicular glutamate transporter 2 (VGLUT2), and 

these probably originate from local excitatory interneurons. This suggests that GluR4containing 

AMPArs in the spinal cord may play an important role in pain mechanisms. 



T14 New insights into mechanisms of spinal LTP and pain memory 

Rohini Kuner 

Pharmakologisches Institut, Universität Heidelberg, Im Neuenheimer Feld, 

Heidelberg, 69120 Germany 

The second messenger cGMP is a key mediator of sensitization at spinal synapses, 

which constitutes a cellular basis for chronic pain. Because cGMP can activate several 

ion-channels and enzymes pre- as well as post-synaptically, the precise identity of 

critical signaling molecules, their mechanisms of action and their locus in the spinal 

circuitry have remained unclear. I will present some results which indicate that the 

cGMP-dependent protein kinase 1 (PKGI) localized presynaptically in nociceptor 

terminals mediates long term potentiation at spinal synapses by enhancing 

presynaptic vesicular release. In nociceptors, we observed that PKGI produces 

activity-dependent phosphorylation of proteins, which directly regulate intracellular 

calcium signaling and vesicular release and set the excitation thresholds of painsensing 

nerves. Our results indicate that PKGI localized presynaptically in nociceptors 

is the most important spinal target of cGMP and represents a key convergence point 

of the NMDA-NO-soluble guanylyl cyclase pathway and the natriuretic peptidemembrane 

guanylyl cyclase pathway in mediating nociceptor sensitization and chronic 

pain. 



T16 Transcriptional Control in Dorsal Spinal Cord Development: New Roles for 

Old Factors 

Jane Johnson 

University of Texas Southwestern Medical Center 

The dorsal horn of the spinal cord contains the neuronal circuitry that modulates 

sensory input from the periphery. The formation of this circuitry, including the balance 

of inhibitory and excitatory neuronal subtypes, is initially generated through 

specification mechanisms controlled by transcription factors, particularly members of 

the homeodomain (HD) and the basic helix-loop-helix (bHLH) families. bHLH 

transcription factors such as Ascl1 (previously Mash1), Atoh1 (previously Math1) and 

Neurog1/2 (previously Ngn1/2) are in a balance with Notch signaling to regulate the 

number of progenitor cells undergoing neuronal differentiation as determined by cell 

cycle exit, movement out of ventricular zones, and expression of neuronal specific 

markers. In the developing dorsal spinal cord, these same bHLH factors, and related 

family members, function in neuronal subtype specification as well. For example, Ptf1a 

is essential for the generation of GABAergic inhibitory neurons in multiple regions of 

the nervous system. In the absence of Ptf1a, dorsal horn GABAergic neurons are misspecified 

to glutamatergic neurons. Probing deeper into the mechanism of Ptf1a 

function, the existence of a novel DNA binding complex was uncovered. This complex 

contains Ptf1a and its E-protein partner, plus Rbpj—the transducer of the Notch 

signaling pathway. We demonstrate the Ptf1a--Rbpj interaction is required in vivo for 

specification of the GABAergic neurons, a function that cannot be substituted by the 

classical form of the bHLH heterodimer with E-protein or Notch signaling through Rbpj. 

Thus, this unique Ptf1a-Rbpj complex controls the balanced formation of inhibitory and 

excitatory neurons in the developing spinal cord, and reveals a novel Notch 

independent function for Rbpj in nervous system development. The interplay between 

bHLH transcription factors and Notch signaling components occurs at multiple levels 

and is key in directing both neuronal differentiation and neuronal subtype specification. 



T17 The Novel Estrogen Receptor GPR30 Mediates Estrogen-Induced And 

PKCepsilon -Dependent Mechanical Hyperalgesia In Vitro And In Vivo 

Julia Kuhn 1 , Olayinka Dina 2 , Jon Levine 2 , Tim Hucho 1 

1 Max Planck Institut für molekulare Genetik, 2 University of California, San Francisco 

The epsilon isoform of protein kinase C (PKCepsilon) is an important second 

messenger in models of acute as well as chronic mechanical sensitization. Only few 

receptors have been identified to lead to activation of PKCepsilon (beta2-adrenergic 

receptor, bradykinin receptor). Recently, we reported estrogen to activate PKCepsilon 

in primary sensory neurons as well as induce PKCepsilon-dependent sensitization in 

male rats. Which of the known estrogen receptors (estrogen receptor alpha (ERalpha), 

estrogen receptor beta (ERbeta), G-protein coupled receptor 30 (GPR30)) mediates 

this effect remained unknown. Here we show, that the agonist of the novel estrogen 

receptor GPR30, G-1, induces rapid PKCepsilon-translocation in primary nociceptive 

neurons. Also, ICI 182 780, which acts as an agonist of GPR30 while blocking the 

classical estrogen receptors ERalpha and ERbeta, leads to fast activation of 

PKCepsilon. In contrast, the specific agonists of ERalpha, PPT, and ERbeta, DPN, do 

not have any effect. RT-PCR studies show, indeed, GPR30 to be expressed in rat 

dorsal root ganglia. Also, in behavioural experiments intradermal injection of G-1 into 

the dorsum of the paw of male rats induces strong mechanical hyperalgesia in a 

concentration dependent manner. These data indicate an involvement of the novel 

estrogen receptor GPR30 in the modulation of nociceptive signaling pathways by sex 

hormones. 



T18 Adaptations in the C Fiber Nociceptors of Naked Mole-Rats Render These 

Animals Insensitive to Specific Air-Borne Irritants and CO2-Induced 

Pulmonary Edema 

Thomas Park 1 , Gary Lewin 2 

1 Laboratory of Integrative Neuroscience, Department of Biological Sciences, 

University of Illinois at Chicago, Chicago, Illinois, USA., 2 Max-Delbrück Center for 

Molecular Medicine, Robert-Rössle-Str. 10, Berlin-Buch D-13092 Germany and 

Charité-Universitätsmedizin, Berlin. 

African Naked mole-rats have a unique ecology in that they are fully subterranean and 

they live in very high numbers. Hence, the air in their poorly ventilated living space has 

chronically high concentrations of CO2 and ammonia. In the respiratory tract these 

chemical irritants activate C fiber nociceptors (also referred to as chemoreceptors or 

irritant detectors). We have identified two putative adaptations in the C fiber 

nociceptors of naked mole-rats that are consistent with evolving under the challenging 

conditions this species faces. First, their C fibers lack the neuropeptides Substance P 

(SP) and calcitonin gene related peptide (CGRP), and second, the fibers are 

unresponsive to low pH (acidosis). We hypothesized that these adaptations would 

render the mole-rats behaviorally insensitive to chemical irritants that act on trigeminal 

C fibers (high CO2, ammonia, capsaicin), and physiologically resistant to the effects of 

lung acidosis associated with breathing high levels of CO2 (neurogenic inflammation 

and pulmonary edema). Here we report that naked mole-rats do not respond to 

capsaicin applied to the nasal cavity while the same concentration of capsaicin 

induces vigorous nose wiping in mice. In an avoidance test, we found that the naked 

mole-rats do not avoid fumes from ammonia or acetic acid. They do, however, avoid 

fumes from nicotine which acts on trigeminal Aδ fibers, not C fibers. Laboratory rats 

avoided all three irritants. To test the effects of high levels of CO2 on the lungs, we 

exposed naked mole-rats and mice to a variety of CO2 concentrations while holding 

O2 constant. Mice showed significant pulmonary edema at concentrations of 15% 

CO2 and higher. The mole-rats showed no edema even at 50% CO2. These results 

support the idea that the anomalies found in naked mole-rat C fibers represent 

adaptations for surviving in chronically high levels of CO2 and ammonia. Experiments 

that exploit the unique features of the naked mole-rat, like those presented here, are 

helping us better understand basic trigeminal and pulmonary physiology as well as 

adaptations to challenging environments. 


T19 Tactile Experience Shapes Behavior in Etruscan Shrews 

Michael Brecht, Farzana Anjum 

Bernstein Center for Computational Neuroscience, Berlin 


A crucial role of tactile experience for the maturation of neural response properties in 

the somatosensory system is well established, but little is known about the role of 

tactile experience in the development of tactile behaviors. Here we study how tactile 

experience affects prey capture behavior in Etruscan shrews, Suncus etruscus. Prey 

capture in adult shrews is a high-speed behavior that relies on precise attacks guided 

by tactile Gestalt cues. We studied the role of tactile experience by three different 

approaches. First, we analyzed the hunting skills of young shrews right after weaning. 

We found that prey capture in young animals is most but not all aspects similar to that 

of adults. Second we performed whisker trimming for three to four weeks after birth. 

Such deprivation resulted in a lasting disruption of prey capture even after whisker regrowth: 

attacks lacked precise targeting and had a lower success rate. Third, we 

presented adult shrews with an entirely novel prey species, the giant cockroach. The 

shape of this roach is very different from the shrew’s normal (cricket) prey and the 

thorax – which is the preferred point of attack in crickets – is protected a heavy cuticle. 

Initially shrews attacked giant roaches the same way they attack crickets and targeted 

the thoracic region. With progressive experience, however, shrews adopted a new 

attack strategy targeting legs and underside of the roaches and only rarely other body 

parts. Speed and efficiency of attacks improved. These data suggest that tactile 

experience shapes prey capture behavior. 



T20 Specification and differentiation of dorsal spinal cord interneurons. 

Thomas Müller 1 , Hendrik Wildner 1 , Domenique Bröhl 1 , Mathias Treier 2 , 

Carmen Birchmeier 1 

1 Max-Delbrück-Center for Molecular Medicine, Berlin, Germany, 2 EMBL, Heidelberg, 

Germany 

The dorsal horn of the spinal cord receives sensory information from the periphery, 

processes this information and relays it to higher brain centers and to motor neurons 

in the ventral spinal cord. Interneurons of the dorsal horn are organized in laminae, 

which receive sensory input of characteristic modalities. For instance, nociceptive 

sensory fibers synapse in superficial laminae, whereas proprioceptive sensory fibers 

project into the deep dorsal horn. Interneurons in the dorsal horn have diverse 

physiological characteristics. Their molecular characteristics are only now beginning to 

be understood. We have focused on the analysis of molecular mechanisms that are 

responsible for the specification of different neuron types of the dorsal horn, and 

analyzed a number of transcription factors that control development of dorsal neurons. 

Recent data on this work will be discussed. Hori, K., J. Cholewa-Waclaw, et al. (2008). 

"A nonclassical bHLH Rbpj transcription factor complex is required for specification of 

GABAergic neurons independent of Notch signaling." Genes Dev 22(2): 166-78. 

Zechner, D., T. Muller, et al. (2007). "Bmp and Wnt/beta-catenin signals control 

expression of the transcription factor Olig3 and the specification of spinal cord 

neurons." Dev Biol 303(1): 181-90. Sieber, M. A., R. Storm, et al. (2007). "Lbx1 acts 

as a selector gene in the fate determination of somatosensory and viscerosensory 

relay neurons in the hindbrain." J Neurosci 27(18): 4902-9. Wildner, H., T. Muller, et 

al. (2006). "dILA neurons in the dorsal spinal cord are the product of terminal and nonterminal 

asymmetric progenitor cell divisions, and require Mash1 for their 

development." Development 133(11): 2105-13. Muller, T., K. Anlag, et al. (2005). "The 

bHLH factor Olig3 coordinates the specification of dorsal neurons in the spinal cord." 

Genes Dev 19(6): 733-43. Zechner, D., Y. Fujita, et al. (2003). "beta-Catenin signals 

regulate cell growth and the balance between progenitor cell expansion and 

differentiation in the nervous system." Dev Biol 258(2): 406-18. Muller, T., H. 

Brohmann, et al. (2002). "The homeodomain factor lbx1 distinguishes two major 

programs of neuronal differentiation in the dorsal spinal cord." Neuron 34(4): 551-62. 



T21 The menthol receptor TRPM8 is the principal detector of environmental cold 

Jan Siemens 1 , Diana M. Bautista 1 , Josh Glazer 2 , Pamela R. Tsuruda 1 , 

Allan I. Basbaum 3 , Cheryl L. Stucky 2 , Sven-E. Jordt 4 , David Julius 1 

1 Departments of Physiology and Cellular & Molecular Pharmacology, University of 

California, San Francisco, CA 94158, USA, 2 Department of Cell Biology, Neurobiology 

and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA, 

3 Departments of Anatomy and Physiology and W.M. Keck Center for Integrative 

Neuroscience, University of California, San Francisco, CA 94158, USA, 4 Department 

of Pharmacology, Yale University School of Medicine, New Haven, CT 06520, USA 

Sensory nerve fibers can detect changes in temperature over a remarkably wide 

range, a process that has been proposed to involve direct activation of 

thermosensitive excitatory TRP ion channels. One such channel, TRPM8 or CMR1, is 

activated by chemical cooling agents (such as menthol) or when ambient 

temperatures drop below ~26°C, suggesting that it mediates the detection of cold 

thermal stimuli by primary afferent sensory neurons. However, some studies have 

questioned the contribution of TRPM8 to cold detection or proposed that other 

excitatory or inhibitory channels are more critical to this sensory modality in vivo. Here 

we show that cultured sensory neurons and intact sensory nerve fibers from TRPM8deficient 

mice exhibit profoundly diminished responses to cold. These animals also 

show clear behavioral deficits in their ability to discriminate between cold and warm 

surfaces, or to respond to evaporative cooling. At the same time, TRPM8 mutant mice 

are not completely insensitive to cold as they avoid contact with surfaces ≤10°C, albeit 

with reduced efficiency. Thus, our findings demonstrate an essential and predominant 

role for TRPM8 in thermosensation over a wide range of cold temperatures, validating 

the hypothesis that TRP channels are the principal sensors of thermal stimuli in the 

peripheral nervous system. 



T22 PAIN ATTENUATION IN TETHERED-TOXIN TRANSGENIC MICE DUE TO 

REDUCED EXCITABILITY OF SENSORY NEURONS 

Annika Stuerzebecher 1 , Jing Hu 2 , Gary R. Lewin 2 , Inés Ibañez-Tallon 1 

1 Molecular Neurobiology, Max-Delbrück-Centrum, Germany, 2 Molecular Physiology of 

Somatic Sensation, Max-Delbrück-Centrum, Germany 

Toxins derived from venomous animals have been widely employed in neuroscience 

research because of their ability to bind and modulate specific ion channels. However, 

the detailed characterization of the structure and function of receptors in neuronal 

circuits is often hampered by the fact that the action of soluble neurotoxins cannot be 

targeted to specific cells. Our group has developed a new strategy using peptide 

toxins that are tethered to the membrane via a GPI anchor and retain their specific 

action on ion channel subtypes while acting in a cell-autonomous manner. Since 

alterations in the function of voltage gated sodium channels (VGSCs) lead to 

hyperexcitability that causes chronic and neuropathic pain, we wanted to address 

whether this strategy could be used to specifically manipulate these channels in vivo. 

For this purpose, we generated transgenic mice using the bacterial artificial 

chromosome (BAC) of the Nav1.8 VGSC to drive expression of the MrVIA conotoxin. 

This toxin is a potent blocker of this tetrodoxin resistant (TTX-R) channel, which plays 

a major role in pain transduction. Using whole cell patch clamp recording from isolated 

sensory neurons a significant reduction of VGSC currents in nociceptive neurons was 

observed in these mice while mechanoreceptors where not affected, demonstrating 

the cell subtype specificity of this approach. This block was restricted to TTX-R 

currents and not compensated by upregulation of TTX-S currents as was observed in 

Nav1.8 knockout mice (Akopian et al). The Nav1.8 gene is predominantly expressed 

in non-peptidergic nociceptors (IB4+) and consistent with this we observed an 

enhanced block in this cell subpopulation. Behavioral studies revealed a remarkable 

lower sensitivity to noxious cold stimuli in the transgenic toxin mice. Thus these 

studies provide the first proof of function of the BAC transgenic tethered toxin 

approach in mammals and its suitability for developing new therapeutic mouse models 

for pain research. (Supported by the DFG SFB 665) 



T23 Acid-Sensing Ion Channels (ASICs) and pain in the central and peripheral 

nervous system 


Institut de Pharmacologie Moléculaire et Cellulaire (IPMC) CNRS/UNSA UMR 6097, 

660 Route des Lucioles, 06560 Valbonne-Sophia Antipolis, France. 

Acid-sensing ion channels (ASICs) are neuronal voltage-insensitive cationic channels 

activated by extracellular acidification, which can trigger membrane depolarization in 

response to local acidosis. They are expressed in the central nervous system and in 

peripheral sensory neurons, where they have been proposed to sense painful tissue 

acidosis that occurs for instance in ischemic, damaged or inflamed tissue. ASICs have 

been associated with a number of different sensory processes including nociception, 

visual transduction, sour taste perception, hearing functions, and mechanoperception. 

In the recent years, we have been interested in the role of these channels in pain 

perception and modulation. We have shown that peripheral ASIC expression and 

activity are largely increased during inflammation. We have identified several ASICassociated 

proteins and regulatory mechanisms. We have characterized ASICs in the 

spinal cord and we have developed and used a specific toxin blocker to demonstrate 

the implication of the ASIC1a isoform in pain modulation in the central nervous system 

through the opioid system. All these data support a role for ASICs in nociception both 

at the central and peripheral level. Supported by CNRS, INSERM, AFM, ANR and 

ARC/INCa 



T24 Leukocyte-derived and exogenous opioids acting at peripheral opioid 

receptors control neuropathic pain 

Dominika Labuz, Anja Schreiter, Yvonne Schmidt, Alexander Brack, Heike Rittner, 

Halina Machelska 

Anaesthesiologie, Charité - Campus Benjamin Franklin, Berlin, Germany 

Background and aims: Neuropathic pain results from nerve injury that can lead to 

inflammation. Challenging the current view that immune cells act predominately as 

generators of neuropathic pain here we investigate analgesic effects of opioids 

derived from leukocytes in response to application of corticotropin-releasing factor 

(CRF) in neuritis. We also assess whether opioid delivery relative to the primary site of 

nerve injury is critical for efficient peripheral analgesia in neuropathic pain. Methods: 

At 14 days after chronic constriction injury (CCI) we examined the expression of opioid 

receptors, CRF receptors and opioid peptides by immunohistochemistry, quantified 

opioid-containig leukocytes by flow cytometry, and measured nociceptive thresholds 

with von Frey test in wild type, beta-endorphin knockout (END KO) and in mice with 

severe combined immunodeficiency (SCID). CRF was injected at the CCI site alone or 

together with antibodies against opioid peptides, antagonists of CRF- and opioidreceptors, 

or after intraperitoneally injected antibody against intercellular adhesion 

molecule-1 (ICAM-1). Mu-, delta- and kappa-opioid receptor selective agonists 

(DAMGO, DPDPE and U50,488H, respectively) were injected at the CCI site or into 

the paw innervated by the ligated nerve. Results: Opioid receptors and opioidcontaining 

leukocytes co-expressing CRF receptors accumulated at the site of nerve 

injury. Wild type, END KO and SCID mice developed mechanical allodynia in paws 

supplied by the ligated nerve. In wild type mice, but not in END KO mice, CRF injected 

at the CCI site produced local antinociception that was reversed by antibodies against 

beta-endorphin, met-enkephalin and dynorphin, by CRF receptor antagonist, by 

selective mu-, delta- and kappa-opioid receptor antagonists, by a peripheral opioid 

receptor antagonist, and by anti-ICAM-1-mediated attenuation of opioid-containing 

leukocyte accumulation at the injured nerve. In SCID mice, which lack T lymphocytes, 

CRF antinociception was significantly decreased as compared with wild type animals. 

Mu-, delta- and kappa-receptor selective agonists injected either at the CCI site or into 

the paw innervated by the injured nerve dose-dependently attenuated mechanical 

allodynia. However, the antinociceptive potency and efficacy of opioids was greater 

when they were injected at the site of nerve injury than into the paw innervated by the 

ligated nerve. Conclusions: Activation of opioid peptide-containing leukocytes that 

accumulate at the site of nerve injury is critical for alleviation of neuropathic pain. 

Unspecific immunosuppression may limit opioid-mediated beneficial effects of 

neuroinflammation. Further, peripheral opioid antinociception in neuropathic pain is 

more prominent when opioid receptors are activated at the site of inflammatory 

reaction associated with nerve injury. Supported by DFG/KFO 100/2. 


T25 Synaptic Dis-Inhibition in Pathological Pain States 

Hanns Ulrich Zeilhofer 


Institute of Pharmacology and Toxicology, University of Zurich, and Institute of 

Pharmaceutical Sciences, ETH Zurich 

Inflammatory diseases and neuropathic insults trigger signaling cascades, which 

frequently lead to intense and long-lasting pain syndromes in affected patients. Such 

pain syndromes are characterized not only by an increased sensitivity to painful stimuli 

(hyperalgesia), but also by a qualitative change in the sensory perception of other, 

tactile stimuli (allodynia) and the occurrence of spontaneous pain in the absence of 

any sensory input. Long-term potentiation (LTP)-like changes in synaptic transmission 

between nociceptive C-fibers and spino-periaqueductal grey projection neurons as 

well as a loss of inhibitory control by GABAergic and glycinergic spinal dorsal horn 

neurons have repeatedly been proposed as underlying principles. While considerable 

evidence supports a significant contribution of C-fiber LTP to hyperalgesia, such 

monosynaptic plasticity can hardly explain the occurrence of allodynia and 

spontaneous pain. Here, I will focus on mechanisms of synaptic dis-inhibition in 

inflammatory and neuropathic pain and show that pathologically heightened pain 

sensitivity can be reversed by restoring synaptic inhibition with drugs that target 

specific spinal GABA-A receptor subtypes. 



T26 Mechanosensitive ion channels, stomatin-like proteins and molecular 

tethers essential for touch. 

Gary Lewin 

Department of Neuroscience, Max-Delbrück Center for Molecular Medicine 

The skin is our largest sensory surface, which we use to contact the outside world 

through our sense of touch. The molecular mechanisms of mechanoreception in the 

skin are poorly understood. Molecules have been identified in flies and worms that are 

necessary for mechanosensation, but similar molecules have yet to be identified in 

mammals. We have characterized the function of stomatin-like protein 3 (SLP3) a 

protein that is expressed in all dorsal root ganglion neurons. The SLP3 amino acid 

sequence is highly homologous to the original member of this family stomatin and to 

the C.elegans protein MEC-2. Genetic and electrophysiological evidence has 

demonstrated that MEC-2 is essential for the function of mechanotransduction 

channels in the touch receptor neurons in the worm. It has also been postulated that 

MEC-2 forms an integral and essential part of the worm mechanotransduction channel 

complex, which also includes two members of Deg/ENaC family Mec-4, and MEC-10. 

Using SLP3 mutant mice, we find that SLP3 is required for the normal function of a 

large number of skin mechanoreceptors. Thus in SLP3-/- mice around 35% of the 

myelinated sensory afferents in the skin show no mechanosensitivity. We also used 

whole cell patch clamp techniques to directly measure mechanosensitive currents that 

can be activated by sub-micron displacement of the neurites of sensory neurons 

acutely isolated in culture. We found that again in around 30% of the cells recorded 

from SLP3-/- mice no mechanosensitive current could be found. By reintroducing the 

SLP3 gene into SLP3-/- mutant sensory neurons we could rescue the function of ion 

channels that underlie the mechanosensitive current. We also found that when SLP3 

is expressed heterologously in cells together with ASIC channels the gating of ASIC 

by protons was inhibited. Thus SLP3 can interact with and modulate the gating of 

ASIC channels. We have recently found that the mechanosenosesnitive currents that 

can be recorded in isolated sensory neurons are highly dependent on the extracellular 

matrix. I will provide evidence that a molecular tether exists that links 

mechanosensitive channels to the extracellular matrix. The molecular nature of this 

tether is at this point still unclear. In summary mammalian somatic sensory neurons 

probably possess a range of mechanotransduction complexes of varying composition 

and the nature of the proteins involved is at the present time still only poorly 

understood. Supported by the DFG. 


T27 Sensory Neuron Mechanotransduction 

John Wood, Francois Rugiero, Ramin Raouf 

UCL London 


Light tough, a sense of muscle position, and the responses to tissue-damaging levels 

of pressure all involve mechanosensitive sensory neurons that originate in the dorsal 

root or trigeminal ganglia. A variety of mechanisms of mechanotransduction have 

been proposed. These range from direct activation of mechanically-activated channels 

at the tips of sensory neurons to indirect effects of intracellular mediators, or chemical 

signals released from distended tissues or specialized mechanosensory end organs 

We have used mammalian sensory neurons in culture to characterise mechanicallygated 

currents, in an attempt to identify their molecular structure. We will present data 

on potential binding targets of the conopeptide NMB-1 (noxious mechanoblocker-1), 

which blocks slowly adapting mechanosensitive currents in DRG neurons, as well as 

the perception of pressure-evoked pain. 



T28 Breathing with Phox2b 

Jean-François Brunet 

Ecole normale supérieure, Paris, France. 

Visceral sensation —as opposed to somatosensation, the main focus of this 

meeting— conveys to the brain inputs that are mostly unconscious and directly 

relevant to homeostasis: arterial pressure, gaz contents of the blood, chemical 

composition of the alimentary bolus or the presence of toxins in the gut or 

bloodstream. We have shown that most of the neurons involved, in the central and 

peripheral nervous systems, develop under the control of the homeodomain 

transcription factor Phox2b. The anatomical and cellular locus for one of the key 

visceral sensory modalities —the detection of elevated CO2 in the blood or 

hypercapnia— has remained contentious for many years. I will present our 

contribution to the recent evidence that this essential component of the “drive to 

breathe” is insured by a group of neurons in the hindbrain, also depending on Phox2b. 



T29 Roles of Runx1 in controlling nociceptor development and pain behaviors 

Qiufu Ma 

Department of Neurobiology, Harvard Medical School and Department of Cancer 

Biology, Dana-Farber Cancer Institute 

In mammals, the perception of pain is initiated by the stimulation of a heterogeneous 

population of nociceptive sensory neurons, or nociceptors. However, how individual 

subgroups of nociceptors control pain perception is still largely unclear. Here we 

present data suggesting that nociceptors undergoing transient Runx1 expression 

might play a critical role for thermal pain and neuropathic pain. Runx1 is a runt domain 

transcription factor that is initially expressed in most, if not all, embryonic nociceptors, 

but its expression is progressively extinguished in about 50% of nociceptors, most of 

which are peptidergic neurons. Genetic studies show that Runx1 is required for the 

expression of many ion channels/receptors and neurotrophin receptors, including TRP 

class thermal receptors, Na+-gated and ATP-gated channels, Mrgpr class G-protein 

coupled receptors, and the Ret receptor tyrosine kinase. Behavioral studies 

demonstrated that mice lacking Runx1 exhibit severe deficits in thermal and 

neuropathic pain. In this presentation, I will show that Runx1-dependent genes are 

divided into two categories, A and B. Category A genes are expressed in nociceptors 

with persistent Runx1 expression, including TRPM8, TRPA1 (weak Runx1 

expression), TRPC3, MrgprD, SNS2, P2X3, Ret and others. Category B genes are 

expressed in nociceptors with transient Runx1 expression, including TRPV1 (high 

level), MrgprA/B/C, and others. A series of genetic manipulations then suggest that 

category B, but not category A, Runx1 targets may be required for 

thermal/neuropathic pain, implying that nociceptors with transient Runx1 expression 

might play a prominent role for these types of pain. 



T30 Peripheral mechanisms for cold detection in intact and injured tissues 


Instituto de Neurociencias de Alicante Universidad Miguel Hernández-Consejo 

Superior de Investigaciones Científicas, Campus de San Juan, Apdo. correos 18, 

03550 Sant Joan d'Alacant, Alicante (Spain) 

Cold-sensitive nerve terminals of primary sensory neurons detect changes in external 

temperature values within an ample range. Moderate temperature reductions evoke 

cooling sensations whereas higher low temperature values elicit sensations of 

unpleasant cold or pain. Cold-sensitive neurons exhibit specific active and passive 

membrane properties and are equipped with different ionic conductances. TRPM8 and 

background K+ currents, TRPA1, IKD and Ih are variably expressed in the soma and 

peripheral endings of primary sensory neurons and determine the final sensitivity to 

cold and the firing characteristics of specific and non-specific cold-sensitive sensory 

endings. Peripheral axotomy of cold receptor fibers innervating the cornea of the eye 

alters their sensitivity and firing response to cold, suggesting that damage to 

peripheral cold-sensitive nerve terminals modifies the peripheral sensory message 

evoked by cold, thus contributing to cold dysesthesias. 


T31 Genetic Determinants of Pain 



Wolfson Centre for Age-Related Diseases, King’s College London, UK 

Pain sensitivity is known to vary considerably between people. This is true both for 

experimental pain, for instance where a standard force is applied to normal healthy 

tissue, but also for disease-related such as osteoarthritis, where it has long been 

recognised up to 50% of individuals with radiographic indications of disease do not 

report pain, and there is a poor correlation between pain and degree of radiographic 

change. The influence of heritable (genetic) factors on pain sensitivity will be reviewed 

in this talk. Mutations in a few individual genes have recently been demonstrated to 

have a dramatic effect on pain appreciation. The best known example is mutations in 

the trkA gene, a number of which lead to the development of congenital insensitivity to 

pain with anhydrosis (also know as hereditary sensory neuropathy type IV). These 

mutations lead to a failure of small diameter nociceptive neurones to develop. More 

recently mutations in a sodium channel gene normally expressed selectively in 

peripheral pain sensitive neurones (so-called Nav 1.7) have also been found to lead to 

both loss of function (congenital analgesia) and gain of function (erythromyalgia) 

phenotypes. We have recently undertaken a classical twin study to evaluate the 

relative contributions of genetic and environmental factors on responses to painful 

stimuli in human volunteers to a wide variety of pain traits. Statistically significant 

genetic components (varying between 22-55%) were seen for the responses to the 

majority of painful stimuli including sensitivity to heating the skin, areas of secondary 

hyperalgesia brush evoked allodynia following a burn injury and iontophoresis of acid 

solutions. Our study demonstrates the importance of genetic factors in determining 

human experimental pain sensitivity, and opens the way for its use as a phenotype in 

gene discovery. Since experimental pain sensitivity is known to be a predictor for 

pathological pain, our data imply that genetic factors have an important aetiological 

contribution towards clinical pain states. 



43 List of Posters 

List of Posters


P1 DEVELOPMENT OF GENETIC TOOLS FOR SELECTIVE AND REVERSIBLE 

SILENCING OF ION CHANNELS USING NATURAL TOXINS 

Sebastian Auer, Rene Jüttner, Ines Ibanez-Tallon 

P2 Consequences of mPGES-1 deletion on spinal cytokine synthesis and COX- 

2 expression 

Christian Brenneis, Ovidiu Coste, Carlo Angioni, Gerd Geisslinger, Klaus 

Scholich 

P3 Neurod genes are essential for neuropeptide expression and diversification 

of GABAergic neurons in the dorsal spinal cord 

Dominique Bröhl, Michael Strehle, Hagen Wende, Kei Hori, Ingo Bormuth, 

Klaus-Armin Nave, Thomas Müller, Carmen Birchmeier 

P4 Auxiliary subunits and the biophysics of single C. elegans force 

transduction channels 

Austin Brown, Zhiwen Liao, Miriam Goodman 

P5 THE CONTRIBUTION OF TRPA1 AND TRPM8 TO COLD ALLODYNIA AND 

NEUROPATHIC PAIN 

Ombretta Caspani, Sandra Zurborg , Paul Heppenstall 

P6 Relevance of kinin receptors in painful processes: study of B1/B2 knockout 

mice 

Cecile Cayla, Michael Bader, Dominika Labuz, Michael Schäfer, Christoph Stein 

P7 Role of Extracellular Matrix in Sensory Mechanotransduction 

Li-Yang Chiang, Jing Hu, Bettina Erdmann, Manuel Koch, Gary R Lewin 

P8 The transcription factor Rbp-J is required for specification of GABAergic 

neurons in dorsal spinal cord. 

Justyna Cholewa-Waclaw, Hendrik Wildner, Benedetta Martarelli, Carmen 

Birchmeier 

P9 ANTAGONISTIC FUNCTIONAL INTERACTION BETWEEN TRPM8 AND Kv1 

POTASSIUM CHANNELS DETERMINES NOXIOUS COLD DETECTION 

Elvira de la Peña, Rodolfo Madrid, Donovan-Rodriguez Tansy, Carlos Belmonte, 

Felix Viana 

P10 The role of the Tshz1 gene in development of GABA-ergic circuits in the 

central nervous system 

Alistair Garratt 

P11 TRPV4 Biochemically And Functionally Interacts With The Cytoskeleton 

Chandan Goswami, Tim Hucho 

P12 Stimulation of formyl peptide receptor on PMN mediates peripheral opioid 

analgesia 

Dagmar Hackel, Dominika Labuz, Alexander Brack, Heike Rittner 

List of Posters 44


P13 Experimental skin inflammation is impaired by axotomy but promotes the 

regeneration of skin nerves 

Sven Hendrix, Björn Picker, Frank Siebenhaar, Marcus Maurer, Eva M. Peters 

P14 A tether link required for touch 

Jing Hu, Li-Yang Chiang, Bettina Erdmann, Gary Lewin 

P15 Molecular interactions between stomatin-like proteins and acid-sensing ion 

channels 

Julia Jira, Paul Heppenstall 

P16 The zinc finger transcription factor Bcl11a/Ctip1 is essential for neuronal 

differentiation and sensory circuit formation in dorsal spinal cord 

development 

Anita John, Heike Brylka, Pentao Liu, René Jüttner, E. Bryan Crenshaw III, 

Nancy A. Jenkins, Neal G. Copeland, Carmen Birchmeier, Stefan Britsch 

P17 Sequential maturation of sensory neuron mechanotransduction during 

embryonic development 

Stefan G. Lechner, Rui Wang, Henning Frenzel, Gary R. Lewin 

P18 Characterization of the peripheral Osmoreceptor 

Soeren Markworth, Silke Frahm, Ines Ibanez-Tallon, Jens Jordan, Gary R. Lewin 

P19 Analysis of Opioid receptor/K+ channel coupling in sensory neurons 

D. Nockemann, C. Stein P. A. Heppenstall 

P20 Direct inhibition of nociceptors by mu-opioid receptor agonist in 

neuropathic pain 

Yvonne Schmidt, Paul A. Heppenstall, Shaaban A. Mousa, Halina Machelska 

P21 Prevention of opioid peptide degradation for pain control in peripheral 

inflamed tissue 

Anja Schreiter, Carmen Gore, Shaaban A. Mousa, Bernard P. Roques, 

Christoph Stein, Halina Machelska 

P22 TRPC5 channel is activated by osmotically induced membrane stretch 

Sergio Soriano, Carlos Belmonte, Félix Viana, Ana Gomis 

P23 Opioid withdrawal increases TRPV1 activity in a PKA dependent manner 

Viola Spahn, Christian Zoellner 

P24 A molecular dissection of TRPV1 sensitisation using the naked mole-rat 

Ewan St. John Smith, Gireesh Anirudhan, Gary Lewin 

P25 Analysis of phosphorylation targets of cGMP-dependent kinase I involved 

in sensory axon bifurcation 

Agne Stonkute, Hannes Schmidt, Fritz G. Rathjen 

45 List of Posters


P26 New In Vitro mechano-assay reveals distinct intrinsic mechanosensitive 

subtypes in cultured sensory neurons 

Dmitry Usoskin, Per Uhlen, Patrik Ernfors 

P27 Function of Cav3.2 in Sensory neuron mechanosensitivity 

Rui Wang, Gary R. Lewin 

List of Posters 46


47 Poster Abstracts 

Poster Abstracts


P1 DEVELOPMENT OF GENETIC TOOLS FOR SELECTIVE AND REVERSIBLE 

SILENCING OF ION CHANNELS USING NATURAL TOXINS 

Sebastian Auer, Rene Jüttner, Ines Ibanez-Tallon 

Max-Delbrueck-Center for Molecular Medicine, Cellular Neuroscience, Berlin, 

Germany, 

Toxins derived from venomous animals have been widely employed in neuroscience 

research because of their ability to manipulate specific ion channels. However, the 

detailed characterisation of the structure and function of receptors in neuronal circuits 

is often hampered by the fact that the action of soluble neurotoxins cannot be targeted 

to specific cells. Thus we constructed recombinant toxins for the cell-autonomous 

manipulation of neuronal receptors by tethering them to the cell membrane via the GPI 

anchor of the endogenous prototoxin lynx1. In former studies we showed that 

recombinant conotoxins as well as bungarotoxins were not dispersed in solution and 

retained their high specificity for target ion channels, indicating that this approach can 

be used to restrict the site of neurotoxin action to genetically targeted cells. Here, we 

extended our studies to various other toxins derived from snails, spiders, scorpions 

and sea anemones by characterising their action on voltage- and ligand-gated ion 

channels. Furthermore we optimized the cell membrane attachment of the toxins by 

exchanging the GPI anchor with a transmembrane domain fused to the green 

fluorescent marker gene GFP, thus allowing the easy monitoring of the recombinant 

molecules. In a next step we set up both a constitutive as well as an inducible lentiviral 

vector expression system for the efficient in vitro and in vivo transduction of neuronal 

cells with the optimized constructs, allowing the detailed structure-function analysis of 

the ion channels of interest. To sum up, these novel tethered toxins are now optimized 

for in vivo use, thus offering new possibilities for investigations regarding the 

physiology of neuronal circuits. 

Poster Abstracts 48


P2 Consequences of mPGES-1 deletion on spinal cytokine synthesis and COX-2 

expression 

Christian Brenneis, Ovidiu Coste, Carlo Angioni, Gerd Geisslinger, Klaus Scholich 

Institute of Clinical Pharmacology, pharmazentrum frankfurt, ZAFES, Klinikum der 

Johann Wolfgang Goethe-Universität, Theodor-Stern-Kai 7, Frankfurt, Germany 

Cyclooxygenase-2 (COX-2)-dependent prostaglandin (PG) E2 synthesis in the spinal 

cord plays a major role in the development of inflammatory hyperalgesia and 

allodynia. In contrast to its pro-nociceptive effects, PGE2 also limits its own synthesis 

by inhibiting the release of pro-inflammatory cytokines from microglia. Our previous 

work showed that microsomal PGE2 synthase-1 (mPGES-1) -deletion shifts spinal 

COX-2-mediated prostaglandin synthesis from PGE2 to other prostaglandins. Using 

primary spinal cord cultures we demonstrate here that only PGE2 and not PGD2, 

PGF2alpha or the stable prostacyclin-receptor-agonist cicaprost inhibits LPS induced 

tumour necrosis factor alpha (TNFalpha) synthesis, COX-2 and mPGES-1 expression 

and COX-2 activity. This PGE2 effect can be mimicked by EP-2- and EP-4-receptor 

agonists. A multi epitope ligand cartographie (MELC) showed strong EP-2-receptorexpression 

on spinal microglia. Next we determined in an in vivo model whether a 

deletion of mPGES-1 can increase LPS stimulated spinal cytokine synthesis and 

COX-2 expression. The intrathecal injection of LPS resulted in a time-dependent 

increase of TNFalpha concentrations as well as COX-2 expression and activity in 

lumbal spinal cords. In this in vivo model mPGES-1-/- mice exhibited a 80% reduced 

spinal PGE2 synthesis. However, most surprisingly, mPGES-1-/- mice had no 

differences in spinal TNFalpha concentration as well as COX-2-expression and - 

activity as compared to their littermates. The results suggest that in spinal cells 

exogenously applied PGE2 is a potent inhibitor of cytokine synthesis, while in vivo 

mPGES-1 derived PGE2 does not have this effect. 

49 Poster Abstracts


P3 Neurod genes are essential for neuropeptide expression and diversification 

of GABAergic neurons in the dorsal spinal cord 

Dominique Bröhl 1 , Michael Strehle 1 , Hagen Wende 1 , Kei Hori 2 , Ingo Bormuth 3 , 

Klaus-Armin Nave 3 , Thomas Müller 1 , Carmen Birchmeier 1 

1 Max-Delbrück-Centrum for Molecular Medicine, 2 University of Texas Southwestern 

Medical Center, 3 Max Planck Institute of Experimental Medicine 

Neurons in the dorsal horn of the spinal cord receive somatosensory information from 

the periphery, integrate the information and transmit it to higher brain centers. These 

neurons express many different neuropeptides that modulate sensory perception like 

the sensation of pain. Inhibitory neurons of the dorsal horn derive from postmitotic 

neurons that express Pax2, Lbx1 and Lhx1/5, and diversify during maturation. In 

particular, fractions of maturing inhibitory neurons express various neuropeptides. We 

show here that a co-ordinate molecular mechanism determines inhibitory and 

peptidergic fate in the developing dorsal horn. A bHLH factor complex that contains 

Ptf1a acts as upstream regulator and initiates the expression of several downstream 

transcription factors in the future inhibitory neurons, of which Pax2 is known to 

determine the neurotransmitter phenotype. We demonstrate that dynorphin, galanin, 

NPY, nociceptin and enkephalin expression depends of Ptf1a, indicating that these 

neuropeptides are expressed in inhibitory neurons. Furthermore, we show that 

Neurod1/2/6 and Lhx1/5, which act downstream of Ptf1a, control distinct aspects of 

petidergic differentiation. In particular, the Neurod1/2/6 factors are essential for 

dynorphin and galanin expression, whereas the Lhx1/5 factors are essential for NPY 

expression. We conclude that a transcriptional network operates in maturing dorsal 

horn neurons that co-ordinately determines transmitter and peptidergic fate. 

Poster Abstracts 50


P4 Auxiliary subunits and the biophysics of single C. elegans force transduction 

channels 

Austin Brown, Zhiwen Liao, Miriam Goodman 

Dept. of Molecular and Cellular Physiology, Stanford University 

The stomatin-related protein MEC-2 and the paraoxonase-related protein MEC-6 are 

members of the C. elegans touch receptor neuron MEC 4/MEC 10 

mechanotransduction ion channel complex. Coexpression of these auxiliary subunits 

with MEC-4 and MEC-10 in Xenopus oocytes leads to a dramatic and synergistic 

increase in whole-cell current. Using single-channel recordings in outside-out patches, 

we demonstrate that this increase cannot be explained by an increase in open 

probability of active channels or single-channel conductance. We also show that 

coexpression of MEC-2 or MEC-6 does not increase in the amount of MEC-4 protein 

in the plasma membrane. We conclude that in the absence of MEC-2 and MEC-6 the 

vast majority of channels occupy a non-conducting state despite correct expression at 

the plasma membrane. We also are investigating the role of cholesterol in the ion 

channel complex, since MEC 2 exhibits cholesterol-binding activity required for 

behavioral response to touch in vivo. 

51 Poster Abstracts


P5 THE CONTRIBUTION OF TRPA1 AND TRPM8 TO COLD ALLODYNIA AND 

NEUROPATHIC PAIN 

Ombretta Caspani, Sandra Zurborg, Paul Heppenstall 

Clinic for Anesthesiology and Operative Intensive Care, Charté Universitätsmedizin 

Berlin, Germany 

Cold allodynia is a common symptom of neuropathic pain. However the underlying 

mechanisms of this enhanced sensitivity to cold are not known. TRPM8 and TRPA1 

are non-selective cation channels expressed by sensory neurons that have been 

proposed as candidates for cold transducers. We have investigated the role of these 

ion channels in cold allodynia by examining their expression and function following 

nerve injury. We used a chronic constriction injury of the sciatic nerve to model 

neuropathic pain in mice. We dissected lumbar dorsal root ganglia (DRG) at 7 and 14 

days post surgery and used quantitative RT-PCR, in situ hybridization, 

immunohistochemistry and calcium microfluorimetry to examine the expression and 

function of TRPM8 and TPRA1 after nerve injury. With the expression analysis we 

detected decrease in the percentages of the both subpopulations of neurons, TRPA1 

positive and TRPM8 positive, after nerve injury. We then examined functional 

properties of cold transduction using calcium imaging. Our experiments revealed a 

down-regulation of TRPA1 in the DRG after nerve injury, although no change in the 

number of cold responsive neurons. 

Poster Abstracts 52


P6 Relevance of kinin receptors in painful processes: study of B1/B2 knockout 

mice 

Cecile Cayla 1 , Michael Bader 2 , Dominika Labuz 1 , Michael Schäfer 1 , Christoph Stein 1 

1 Anaesthesiologie, Charite, Campus Benjanin Franklin, Berlin, 2 Max-Delbrück-Center, 


Kinins are peptides rapidly produced during inflammation that play a critical role in the 

initiation of pain and the development of hypersensitivity. Kinins act through two 

distinct G-protein-coupled receptors, constitutively expressed B2 and injury-induced 

B1. The B2 receptor has been preferentially associated with acute and the B1 receptor 

with chronic phases of inflammation and pain. To progress in our understanding of the 

function of the kinin system, we aimed to generate mice lacking both B1 and B2 

receptors (B1B2-/-) and to study their nociceptive behaviour. Because of the close 

chromosomal proximity of the B1 and B2 receptor genes, this model could not be 

obtained by simple breeding of single-defective lines. By homologous recombination 

of the B1 receptor gene directly in embryonic stem cells isolated from B2-deficient 

animals we generated the mouse model. B1B2-/- mice are viable, morphologically 

normal and fertile. The deletion of the two genes was verified by Southern blot and the 

absence of receptor mRNAs evidenced in all organs tested. In contrast to wild-type 

(WT) mice, an intraplantar injection of bradykinin did not induce nociceptive responses 

in B1B2-/- mice. We next evaluated the response to intraperitoneal administration of 

0.6% acetic acid, which elicited a significant number of abdominal constrictions in WT, 

but a reduced response in the double knockout line. Mechanical and thermal 

sensitivities in healthy B1B2-/- animals were also tested as well as the development of 

hyperalgesia induced by inflammation or nerve injury. 

53 Poster Abstracts

P7 Role of Extracellular Matrix in Sensory Mechanotransduction 


Li-Yang Chiang 1 , Jing Hu 1 , Bettina Erdmann 1 , Manuel Koch 2 , Gary R Lewin 1 

1 Molecular Physiology of Somatic Sensation Group, Max Delbrück Center for 

Molecular Medicine, Berlin, 2 Center for Biochemistry and Department of Dermatology, 

Medical Faculty, University of Cologne 

Genetic screens carried out in both Caenorhabditis elegans (C. elegans) and 

Drosophila Melanogaster have suggested that extracellular proteins might be essential 

for the transducton of body touch and fly bristle movement respectively (Chung et al. 

2001. Neuron 29: 415-28; Du et al. 1996. Neuron 16: 183-94; Ernstrom & Chalfie. 

2002. Annu Rev Genet 36: 411-53). There is no direct evidence that extracellular 

factors are crucial for the gating of somatic mechanotransduction channels in 

vertebrates. In order to investigate this, we have established co-cultures of sensory 

neurons and skin-derived keratinocytes or fibroblasts. We have asked whether 

different extracellular environments modify the transduction of mechanical stimuli by 

DRG neurons. In this project, we have found the molecular nature of the extracellular 

environment can profoundly modulate mechanosensitive channels in DRG neurons. 

Matrix derived from keratinocytes has profoundly inhibitory effect on mechanosensitive 

RA currents. It has been shown Laminin5 is exclusively expressed in Keratinocytes. 

We found Laminin5 containing matrix can reproduce the inhibitory properties of 

keratinocytes matrix on DRG mechanosensitivity. Future experiments with laminin5 

deficient matrix will allow us to determine if Lamnin5 itself is sufficient for the inhibitory 

effects of keratinocyte matrix. 

Poster Abstracts 54


P8 The transcription factor Rbp-J is required for specification of GABAergic 

neurons in dorsal spinal cord. 

Justyna Cholewa-Waclaw, Hendrik Wildner, Benedetta Martarelli, Carmen Birchmeier 

Max-Delbruck-Centrum for Molecular Medicine, 13125 Berlin-Buch, Germany 

dILA and dILB neurons, the major neuronal subtypes of the dorsal spinal cord, arise in 

a salt-and-pepper pattern from a broad progenitor domain that expresses Mash1 and 

p48/Ptf1a transcription factors. dILA and dILB differentiate into inhibitory and 

excitatory neurons. Specification of dILA neurons depends on the bHLH transcription 

factor p48/Ptf1a. Furthermore, dILA neurons are produced by asymmetric progenitor 

cell division. To address the role of Notch signaling in the specification of dILA 

neurons, we employed two genetic approaches in mice: (i) expression of a dominantnegative 

variant of Mastermind-like1 (DNMAML) in an inducible manner; Mastermindlike1 

is a transcriptional co-activator that is thought to be essential to activate the 

transcription of Notch-dependent target genes of Rbp-J; (ii) conditional mutagenesis of 

Rbp-J, the major transcriptional mediator of Notch signals. Induction of expression of 

the dominant-negative Mastermind-like1 variant and the introduction of the Rbp-J 

mutation were regulated via cre-recombinase (Pax3Cr or Pax7CreERT2 alleles). 

Compared to control mice, we observed in Pax3Cre/DNMAML mutants a partial 

depletion of the progenitor domain in the dorsal spinal cord at E12.5, consistent with a 

reduction of Notch signaling. However, there was no significant alteration in the 

proportions of dILA and dILB neurons generated. In contrast, we found a significant 

change in the proportions of dILA and dILB neurons in Pax7CreERT2 Rbp-Jflox/flox 

mice. In the pancreas, Rbp-L is known to form a transcription factor complex together 

with p48/Ptf1a. Our results indicate that Rbp-J can function independently of Notch as 

a component of Ptf1a complex in determining the identity of GABAergic neurons. 

Supporting results were obtained from analysis of a mutant of p48/Ptf1a 

(Ptf1aW298A) that encodes a variant with a single change in the amino acid 

sequence. This mutation precludes an interaction between Ptf1aW298A and Rbp-L or 

Rbp-J. Ptf1aW298A is a phenocopy of the Ptf1anull mutant, and inhibitory dILa 

neurons in the dorsal spinal cord are not formed in either of the two mutant mouse 

strains. 

55 Poster Abstracts


P9 ANTAGONISTIC FUNCTIONAL INTERACTION BETWEEN TRPM8 AND Kv1 

POTASSIUM CHANNELS DETERMINES NOXIOUS COLD DETECTION 

Elvira de la Peña, Rodolfo Madrid, Donovan-Rodriguez Tansy, Carlos Belmonte, 

Felix Viana 

Instituto de Neurociencias. Universidad Miguel Hernández-CSIC. Campus de San 

Juan. Alicante-España. 

Detection of temperature decreases in the innocuous range is carried out by a specific 

functional class of peripheral sensory nerve endings named cold thermoreceptors. 

The somas of cold-sensitive primary sensory neurons cultured in vitro present very 

different temperature thresholds (range 34─20ºC) and can be separated into low 

threshold (LT) and high threshold (HT) types. TRPM8, a non-selective, calciumpermeable, 

cation channel of the transient receptor potential superfamily, that is 

activated by cooling and menthol is the principal transduction channel for cold 

temperatures in primary sensory neurons. However, additional experimental evidence 

has shown that various potassium conductances also exert a strong influence on cold 

sensing. We investigated the influence of TRPM8, a cold- and menthol-sensitive TRP 

channel, and IKD, a slowly-inactivating, 4-AP-sensitive transient potassium current on 

temperature-evoked responses in cold thermoreceptors. We performed calcium 

imaging and patch-clamp recordings from neonatal mice cultured trigeminal neurons. 

Temperature response threshold was measured on the cold-evoked intracellular 

calcium elevation. Expression of TRPM8 was quantified by the amplitude of the 

menthol-evoked current (Imenthol). We found that Imenthol was significantly larger in 

LT CS neurons while the opposite was true for IKD. Using pharmacological tools we 

demonstrate that these two conductances exert opposite influences on temperaturedependent 

excitability in trigeminal thermoreceptors. One current, IKD, acts as a brake 

while the other, TRPM8, promotes cold-induced activity. Alterations in the balance 

between ITRPM8 and IKD may be an important factor in the development of cold 

allodynia symptoms, characteristic of many neuropathic conditions. Supported by 

funds from the Spanish Ministry of Education and Science, projects SAF2004-01011 

to F.V , BFU2005 08741 to C.B and Consolider 2007 CSD2007-0023. Fundación 

Marcelino Botín. 

Poster Abstracts 56


P10 The role of the Tshz1 gene in development of GABA-ergic circuits in the 

central nervous system 


Department of Neurosciences, Max-Delbrück-Center for Molecular Medicine, Berlin 

We first characterized the three Tshz genes as being highly expressed in the 

substantia gelatinosa, the first relay station for pain sensation. We chose to analyse 

the function of the Tsh1 gene, which we had shown also to be expressed in cells of 

the outer granule cell layer of the embryonic olfactory bulb. The early development of 

the olfactory bulb is still poorly understood. Our aim was to determine the function of 

Tsh1, a potential integrator of signaling downstream of cell surface receptors (e.g. 

Wnt/beta-catenin signaling) and a partner for Hox-family transcription factors in 

olfactory bulb development. The methods employed were classical mutagenesis in the 

mouse, together with immunohistochemistry, in situ hybridisation and microarray 

analysis. Tsh1 was expressed in an early emigrating population of GABA-ergic 

interneurons, generated in the rostral telencephalon around E11-E12. In Tsh1 

homozygous null embryos, mutant cells clumped together and failed to distribute 

radially within the olfactory bulb, a phenotype associated with changes in Semaphorin 

signaling. In addition, the Tsh1 mutant cells failed to activate expression of GABAergic 

markers such as GAD67 and GABA. Conclusions: Tsh1 is essential for the 

correct radial migration and differentiation of a pioneer population of GABA-ergic 

granule cell neurons within the olfactory bulb. Tsh1 mutant cells failed to express 

Semaphorin 3c and a signaling mediator cypin, which may account for the aberrant 

clumping of mutant cells within the center of the olfactory bulb. 

57 Poster Abstracts


P11 TRPV4 Biochemically And Functionally Interacts With The Cytoskeleton 

Chandan Goswami, Tim Hucho 

Max Planck Institute of molecular Genetics 

TRPV4 and the cytoskeleton have been reported to influence the development of 

mechanical hyperalgesia. If and how these molecules interact is unknown. We now 

describe TRPV4 to form a Ca2+-sensitive complex with components of the tubulin-, 

actin- and neurofilament-cytoskeleton and and CamKII. The C-terminus of the 

nociceptive signaling molecules PKCε TRPV4 is sufficient for complex formation. The 

interaction with soluble and filamentous tubulin and actin is direct. Actin and tubulin 

bind with high affinity and compete for binding. The presence of TRPV4 strongly 

increases the amount of microtubules formed even in presence of microtubule 

depolymerising agents. Accordingly, expression of TRPV4 induces striking 

morphological changes such as formation of lamellipodia, filopodia, as well as neuritelike 

structures. These changes are not restricted to neuron-derived cells but also occur 

in transfected HaCat, ChoKI, and HeLa cells. TRPV4 co-localizes with actin and 

tubulin both in fixed as well as live cells and stabilizes microtubules at membranous 

regions. Activation of TRPV4 induces rapid depolymerization of microtubules. This is 

accompanied by elongation of filopodial structures and by a transition of lammelipodial 

to filopodial structures. Therefore, cell boundaries and growth cone structures rapidly 

retract. Accordingly, rat primary sensory neurons do not extend neurites if cultured in 

the presence of low doses of TRPV4 agonists. This phenotype is restricted to a subset 

of nociceptive neurons, which binds to isolectin B4. 

Poster Abstracts 58


P12 Stimulation of formyl peptide receptor on PMN mediates peripheral opioid 

analgesia 

Dagmar Hackel, Dominika Labuz, Alexander Brack, Heike Rittner 

Department of Anesthesiology and Intensive Care, Berlin, Germany 

Hyperalgesia can be elicited by tissue destruction e.g. in inflammation. Both proalgesic 

(e.g. cytokines, protaglandins, protons) and analgesic (e.g. opioid peptides) 

mediators are secreted during the inflammatory process. Opioid peptides bind to 

opioid receptors on peripheral nerve endings and thereby elicit potent analgesia. 

Polymorphonuclear cells (PMN) are the major source of opioid peptides and are 

important for endogenous analgesia especially during early inflammation. PMNs 

express the formyl peptide receptor recognizing formylated peptides from bacteria or 

mitochondria. The presented work shows that formylated peptides and their receptor 

influence release of opioid peptides in vitro and in vivo. Intraplantar injection of fMLP 

(N-formyl-MET-LEU-PHE), an agonist of FPR, cause an analgesic effect in rats with 

two hours complete Freund's adjuvant (CFA) induced inflammation in the hind paw. 

This analgesia abolishes after depletion of PMNs or after additional intraplantar 

injection of either the opioid receptor antagonist naloxone or antibodies against opioid 

peptides. Moreover PMNs release opioid peptide after fMLP treatment in vitro. The in 

vivo and in vitro effects of fMLP are blockable with t-BocPLPLP (tert-butoxycarbonyl- 

Phe-Leu-Phe-Leu-Phe) or Cyclosporin H which are competitive antagonists of FPR. 

These results reveal that the FPR on PMNs is important for the release of opioid 

peptides and endogenous analgesia during the inflammatory processes. This work 

was supported by the DFG (German Research Foundation). 

59 Poster Abstracts


P13 Experimental skin inflammation is impaired by axotomy but promotes the 

regeneration of skin nerves 

Sven Hendrix 1 , Björn Picker 1 , Frank Siebenhaar 2 , Marcus Maurer 2 , Eva M. Peters 3 

1 Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité – 

Universitätsmedizin Berlin, Germany , 2 Dept. of Dermatology, Charité – 

Universitätsmedizin Berlin, Germany, 3 Neuroscience Research Center, Charité – 

Universitätsmedizin Berlin, Germany 

Purpose: Previously, we have demonstrated that neuronal plasticity in skin is 

substantially influenced by inflammatory mediators. Here, we tested the hypothesis 

whether experimental inflammation may be a useful tool to promote the regeneration 

of skin nerves after lesion. Methods: Skin was denervated by axotomy of all skin 

nerves, which innervate the dorsal back skin of mice. Sham-operated mice without 

axotomy were used as controls. Experimental dermatitis was induced either by 

capsaicin or by sensitizing mice with ovalbumin and complete Freund’s adjuvant to 

provoke a contact dermatitis-like skin inflammation or with ovalbumin and aluminium 

hydroxide to provoke an atopic dermatitis-like skin inflammation. Passive cutaneous 

anaphylaxis (PCA) was induced by specific IgE anti-albumin-dinitrophenol followed by 

dinitrophenol-HAS. Mast cell activation was determined by measuring ear swelling and 

Evans blue extravasation. The numbers of immune cells and nerve fibers were 

determined by standard immunohistochemistry: Mast cells (FITC-avidin, GIEMSA), 

macrophages (F4/80), T cells (CD3), nerve fibers (PGP9.5, GAP-43). Results: Skin reinnervation 

in all compartments was detectable at day 28 after axotomy. Capsaicininduced 

inflammation and PCA was significantly reduced after axotomy. Injection of 

inhibitors of the neuropeptides Substance P and Calcitonin gene-related peptide also 

substantially reduced mast cell-dependent inflammation. Axotomy also significantly 

reduced T cell and macrophage infiltration in atopic and contact dermatitis skin. 

However, in contact dermatitis the remaining inflammation was sufficient to increase 

significantly the number of regenerating nerve fibers in the epidermis, dermis and 

subcutis as well as in selected hair follicle compartments. Conclusions: These data 

suggest that mast cell-dependent dermatitis is suppressed by axotomy, while mast 

cell-independent skin inflammation is sufficient to promote the regeneration of skin 

innervation after lesion. 

Poster Abstracts 60


P14 A tether link required for touch 

Jing Hu, Li-Yang Chiang, Bettina Erdmann, Gary Lewin 

MDC 

Genetic analysis of touch sensation in C. elegans and research on mechanosensitive 

hair cell in inner cochlea have suggested the existence of an extracellular link which 

tethers the mechanosensitive ion channels to matrix and transduces mechanical force 

to the channels (Ernstrom & Chalfie. 2002. Annu Rev Genet 36: 411-53; Syntichaki & 

Tavernarakis. 2004. Physiol Rev 84(4) 1097-153). So far there is no evidence 

demonstrating that an extracellular link is essential for mechanotransduction (Lewin & 

Moshourab. 2004. Neurobiol 61(1): 30-44). Recent evidence suggests that 

mechanotransduction components and mechanisms may be conserved between C. 

elegans and mammals (Wetzel, Hu et al. 2007. Nature 445(7124): 206-9). To address 

the question whether an extracellular link is required in mouse mechanosensation, we 

have established co-cultures of sensory neurons with mouse 3T3 cells (mouse 

embryonic fibrbroblast cell line). We found that stimulating the fibroblast cell adjacent 

to the neurite almost always (94%) evoked fast mechanically gated currents with 

latencies and activation kinetics indistinguishable from those found by stimulating the 

neurite directly. This indicates that there is likely a physical attachment between ion 

channels in the neurite and the fibroblast cell that is required for normal gating of the 

mechanically sensitive current. We have also found that one type of mechanically 

activated current-the rapidly adapting current (RA) is exclusively abolished by a serine 

endopeptidase-Subtilisin. This ablation of mechanosensitivity could be totally 

recovered after further incubating the cells for 30 hrs in culture medium. Using 

transmission electron microscopy (TEM) to visualize the ultrastructure of the 

attachment between cultured neurons and the underlying substrate, we identified a 

novel link protein (length 104.1 ± 3.1nm) tethering sensory neuronal neurites to 

extracellular matrix. After Subtilisin treatment we have found this tether link was 

missing. And it reappeared after 30 hours incubation coincident with the reappearance 

of RA mechanosensitive currents. This is the first evidence that an extracellular link 

required for vertebrate mechanotransducer function exists. We conclude that a long 

extracellular tether link plays an essential role in mouse sensory 

mechanotransduction. 

61 Poster Abstracts


P15 Molecular interactions between stomatin-like proteins and acid-sensing ion 

channels 

Julia Jira, Paul Heppenstall 

Klinik fuer Anaesthesiologie, Charite CBF 

Stomatin-like proteins (SLPs) are widely expressed across species and tissues. 

However, the function of this family of membrane-associated proteins remains to be 

elucidated. Some SLPs bind cholesterol as well as actin filaments and are therefore 

considered to have a scaffolding function, assembling protein complexes at the 

plasma membrane. Moreover, there is evidence of SLPs directly interacting with ion 

channels. In the nematode worm C.elegans, the stomatin homolog MEC-2 plays an 

essential role in mechanosensation by anchoring the transduction channel MEC-4 to 

the cytoskeleton. We are interested in defining the nature of the interactions between 

mammalian counterparts of the C.elegans proteins, which have been implicated in 

mechanotransduction as well. To this end, we have performed co-immunoprecipitation 

and FRET experiments to examine interactions between mouse stomatin-like protein 3 

(mSLP3) and members of the acid-sensing ion channel (ASIC) family. These are 

sodium channels that are homologous to the C. elegans MEC-4 channel and comprise 

short intracellular N- and C-termini, two transmembrane domains and a large 

extracellular loop. All ASIC subunits (ASIC1a, 1b, 2a, 2b, 3 and 4) and mSLP3 were 

epitope tagged and transiently transfected into two cell lines (COS-7 and CHO) that 

are thought to contain no endogenous ASICs. We found that mSLP3 did 

immunoprecipitate with all ASICs. Interestingly, a splice variant of ASIC3, which 

misses the C-terminus and the second transmembrane domain, also interacted with 

mSLP3. This suggests that the interaction site is located more towards the N-terminus 

of the ASIC channels. 

Poster Abstracts 62


P16 The zinc finger transcription factor Bcl11a/Ctip1 is essential for neuronal 

differentiation and sensory circuit formation in dorsal spinal cord 

development 

Anita John 1 , Heike Brylka 1 , Pentao Liu 2 , René Jüttner 3 , E. Bryan Crenshaw III 4 , 

Nancy A. Jenkins 5 , Neal G. Copeland 5 , Carmen Birchmeier 3 , Stefan Britsch 1 

1 Institute for Molecular and Cellular Anatomy, Ulm University, 89081 Ulm, Germany, 

2 The Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK, 3 Max Delbrück Center 

for Molecular Medicine (MDC), 13125 Berlin-Buch, Germany, 4 Mammalian 

Neurogenetics Group, Center for Childhood Communication, The Children's Hospital 

of Philadelphia, PA 19104, USA, 5 Mouse Cancer Genetics Program, Center for 

Cancer Research, National Cancer Institute, Frederick, Maryland 21702, USA 

There is emerging evidence that common regulatory mechanisms are utilized in the 

control of neurogenesis and the development of the lympho-hematopoetic system. 

The zinc finger transcription factor Bcl11a (Evi9, CTIP1) is expressed in 

lymphohematopoetic tissues and in the nervous system. Bcl11a is essential for normal 

lymphoid development, and Bcl11a null mice lack mature B-lymphocytes. Here we 

demonstrate that Bcl11a is essential for neuronal development as well. Conditional 

ablation reveals that dorsal spinal neurons require Bcl11a for terminal differentiation 

and morphogenesis. Moreover, TrkA positive sensory afferents depend in their ability 

to grow into the dorsal horn and to provide synaptic input on the expression of Bcl11a 

in postsynaptic spinal target neurons. A genome-wide screen for transcriptional 

targets down-regulated in Bcl11a mutant spinal neurons identified genes linked to the 

regulation of cytoskeletal dynamics, as well as secreted factors with established 

functions in differentiation and guidance of neurons. Together, the genetic analysis of 

Bcl11a reveals, for the first time, essential functions of this factor in central nervous 

system development. Our data suggest that Bcl11a is required to orchestrate key 

developmental events, which ultimately lead to the establishment of functional 

neuronal circuits. 

63 Poster Abstracts


P17 Sequential maturation of sensory neuron mechanotransduction during 

embryonic development 

Stefan G. Lechner, Rui Wang, Henning Frenzel, Gary R. Lewin 

Max Delbrück Centrum f. Molekulare Medizin 

We have reported the presence of three types of mechanically activated currents in 

adult mouse dorsal root ganglion (DRG) neurons. These currents can readily be 

distinguished by their inactivation kinetics and are thus classified into rapidly adapting 

(RA-type), intermediately adapting (IA-type) and slowly adapting (SA-type) currents. In 

the present study we asked when during embryonic development do sensory neurons 

acquire these mechanosensitive currents. Our data shows that mechanosensitive 

currents emerge in three major waves that coincide with already well-described 

epochs in the development of the sensory ganglia. First, at E13.5, mechanoreceptors 

and proprioceptors acquire RA-type currents, followed by nociceptors which start to 

acquire RA-currents at E15.5. The third wave of mechanosensitivity occurs just after 

birth when a large proportion of nociceptors acquire an SA-type current. In addition we 

investigated the electrical properties of developing sensory neurons. We found that 

both, mechanoreceptors and nociceptors, become electrically excitable immediately 

after they are born, at E11.5 and at E12.5-E13.5, respectively. Thus the acquisition of 

electrical excitability precedes that of mechanosensitivity by approximately two days. 

Moreover, we found that the emergence of mechanically gated currents in nociceptors 

correlates with significant changes in the electrical excitability of these neurons. Since 

the first two waves coincide with the innervation of peripheral targets by 

mechanoreceptors (E13.5) and nociceptors (E15.5), we next asked whether target 

derived factors are required for the acquisition of mechanosensitivity. Therefore DRG 

neurons from non-mechanosensitive stages were differentiated in-vitro in the 

presence of different neurotrophins. These experiments strongly suggest that the 

acquisition of mechanosensitive currents by nociceptors is induced by target derived 

NGF, whereas the acquisition of RA-currents by mechanoreceptors may be a timedependent 

process which is independent of the presence of neurotrophic factors. 

Supported by SFB 665 

Poster Abstracts 64


P18 Characterization of the peripheral Osmoreceptor 

Soeren Markworth 1 , Silke Frahm 1 , Ines Ibanez-Tallon 1 , Jens Jordan 2 , Gary R. Lewin 1 

1 Molecular Physiology of Somatic Sensation, Department of Neuroscience, 

Max-Delbrück Center for Molecular Medicine, Robert-Rössle Str. 10, Berlin-Buch, 

2 Franz-Volhard Centrum für Klinische Forschung, Haus 129, Charité Campus Buch, 

Wiltbergstr. 50, Berlin-Buch 

Osmosensitive fibers are believed to be located in the liver and/or the hepatic branch 

of the portal vein and project to the CNS via hepatic nerves. Peripheral osmosensitive 

fibers have not yet been visualized or functionally characterized, indeed the location 

and sensory origin of such fibers is not known. Here we have studied the origin as well 

as the activation mechanism of hepatic osmosensitive nerve fibers. We show that in 

rats and mice drinking water leads to a decrease of blood osmolality in the portal vein 

by 25mOsm. We visualized activation of hepatic sensory fibers by removing the tissue 

shortly after the stimulus and immunostaining for phosphorylated ERK (extra-cellularsignal-related-kinase). 

The osmotic stimulus led to a robust induction of pERK in a 

sub-population of Isolectin B4-negative sensory nerve fibers. We made acute cultures 

of dorsal root ganglion neurons from the thoracic ganglia (T7-T13) a proportion of 

which innervate visceral tissues including the liver. Of these cells 31% show a robust 

increase in intracellular calcium following stimulation with hypotonic solutions (Fura-2 

based calcium imaging). Sensory neurons taken from ganglia that do not innervate 

viscera had a significantly smaller number of osmosensitive cells. The Transient 

Receptor Potential Vanniloid 4 (Trpv4) is believed to play a role in osmosensation. 

Here we show using Ca2+-Imaging and patch clamp techniques a significant decrease 

in osmosensitive cells of Trpv4 -/- animals compared to WT animals only in the 

thoracic, but not in the cervical and lumbar region. We found that only small to 

medium sized cells of Trpv4 -/- animals have lost their ability to respond to osmotic 

challenges. We also examined a transgenic mouse in which the alpha3 subunit of the 

nicotinic acetylcholinreceptor is fused to GFP (green fluorescent protein). We show, 

that ERK becomes activated in this subpopulation of neurons after water intake. The 

percentage of GFP+ cells showing osmosensitivity in ca-imaging experiments was 

much higher than GFP- cells in thoracic ganglia. This animal model to study the 

activation of peripheral osmoreceptors may prove a valuable tool to dissect the 

physiological relevance of this sensory pathway. 

65 Poster Abstracts


P19 Analysis of Opioid receptor/K+ channel coupling in sensory neurons 

D. Nockemann, C. Stein, P. A. Heppenstall 

Opioids are the most effective and widely used drugs in the treatment of severe pain. 

They act through G-protein-coupled receptors located on central and peripheral 

sensory neurons. Three classes of opioid receptors (mu, delta, kappa) have been 

identified. Several mechanisms are involved in opioid analgesia including G-proteingated 

inwardly rectifying K+ (GIRK) channel activation and inhibition of voltage-gated 

Ca2+ channels. Interestingly, in peripheral sensory neurons Ca2+ channel inhibition 

has been the most studied mechanism and GIRK channels have not been considered 

in any detail. The GIRK channels form homo- or heterotetrameric complexes, and are 

known to support the inhibitory effects of opioids in the central nervous system. The 

subunits GIRK1 and 2 have been found to be expressed within the spinal cord dorsal 

horn, the cerebellum, the hippocampus and the cortex. We are interested in whether 

GIRK channels are present and whether opioids are coupled to GIRKs on peripheral 

sensory fibers. Furthermore, we will investigate the role of GIRK/opioid receptor 

coupling in a mouse model of inflammatory and neuropathic pain. To approach these 

questions, we are currently performing quantitative RT-PCR, western blot and patch 

clamping experiments. We found no significant expression of all 4 GIRK subunits in 

sensory neurons using RT-PCR and western blot. No change in the expression level 

could be observed after inflammation of the mouse hindpaw. We also investigate 

functional coupling of GIRK channels and opioid receptors in peripheral sensory 

neurons using Patch clamping. We found no evidence for inward rectifying currents in 

mouse DRG neurons, either before or after treatment with GIRK agonist DAMGO. Our 

data indicate that GIRK channels are not present in sensory neurons and do not 

contribute to peripheral analgesia under normal and inflamed conditions. 

Poster Abstracts 66


P20 Direct inhibition of nociceptors by mu-opioid receptor agonist in 

neuropathic pain 

Yvonne Schmidt, Paul A. Heppenstall, Shaaban A. Mousa, Halina Machelska 

Anaesthesiologie, Charité - Campus Benjamin Franklin, Berlin, Germany 

After nerve injury (e.g. compression, limb amputation) normally innocous tactile 

stimulation (e.g. skin to clothes contact) can result in painful sensation. Currently used 

centrally acting opioids are often associated with CNS-mediated side effects. These 

can be avoided by selective activation of peripheral opioid receptors without affecting 

their analgesic properties. Here we investigate the expression pattern of peripheral 

mu-opioid receptors and the effects of their activation on nociceptor thresholds to 

mechanical stimuli in a mouse model of neuropathic pain. In a chronic constriction 

injury (CCI) of the sciatic nerve we examined the expression of mu-opioid receptor in 

lumbar (L4-L5) dorsal root ganglia (DRG) with immunohistochemistry and a coexpression 

of mu-opioid receptors with different sensory neuronal markers in DRGs 

and in the ligated nerve by double immunofluorescence. Effects of mu-receptor 

agonist DAMGO on von Frey-induced mechanical stimulation were tested in an in vitro 

skin-saphenous nerve preparation. All experiments were performed 2 weeks after 

CCI, sham-surgery or in animals without operation. We found that the number of DRG 

cells expressing mu-opioid receptors did not significantly change after CCI. In DRG 

and in the ligated nerves, mu-opioid receptors were co-expressed with calcitonin 

gene-related peptide and to a lesser degree with isolectin B4 or neurofilament 200, 

markers of peptidergic C and A fibers, non-peptidergic C fibers, and A fibers, 

respectively. The conduction velocity and the threshold to mechanical stimuli in 

sensory afferent fibers between ligated and unligated nerves did not differ significantly. 

In the injured nerve about 50% of Aä and C fibers, but not Aâ fibers, were DAMGOsensitive, 

resulting in an increased mechanical threshold after DAMGO application to 

their receptive field. This effect was reversed by CTOP, suggesting a selective muopioid 

receptor-mediated action. Our results suggest that peripheral mu-opioid 

receptors can be directly activated in cutaneous C and Aä fibers to attenuate their 

excitability after nerve injury. This might constitute a basis for the peripheral opioidmediated 

blockade of mechanical allodynia in neuropathic pain. 

67 Poster Abstracts


P21 Prevention of opioid peptide degradation for pain control in peripheral 

inflamed tissue 

Anja Schreiter 1 , Carmen Gore 1 , Shaaban A. Mousa 1 , Bernard P. Roques 2 , 

Christoph Stein 1 , Halina Machelska 1 

1 Anaesthesiologie, Charité - Campus Benjamin Franklin, Berlin, Germany., 

2 CNRS UMR, Paris, France. 

Background and aims. Degradation of opioids by peptidases such as aminopeptidase 

N (APN) and neutral endopeptidase (NEP) has been extensively examined in the 

central nervous system. In peripheral inflamed tissue immune cells liberate opioid 

peptides, which bind to opioid receptors on sensory nerve terminals to inhibit pain. Our 

aim was to investigate the prevention of opioid peptide degradation by blockade of 

APN and NEP in leukocytes to enhance analgesic activity of opioids in inflammatory 

pain. We used classical inhibitors of NEP (thiorphan) and of APN (bestatin) as well as 

RB3008, a novel dual inhibitor of NEP and APN. Methods. Experiments were 

performed at 4 days after injection of Freund’s adjuvant into one hind paw in rats. 

Analgesic effects of peptidase inhibitors were assessed with the paw pressure test. 

Expression of APN, NEP and opioid peptides in inflamed tissue was analyzed by 

immunohistochemistry. Enzymatic activity of APN and NEP was assessed using 

membranes prepared from immune cells isolated from inflamed paws, and was 

measured spectrometricaly or with radioimmunoassay. Results. Co-injection of 

thiorphan and bestatin or application of RB3008 into inflamed paws produced dosedependent 

analgesia. APN, NEP, ß-endorphin and Met-enkephalin were expressed in 

leukocytes in inflamed tissue. In in vitro immune cell membrane assay APN and NEP 

cleaved their specific exogenously added substrates (Ala-ß-naphthylamin and Suc- 

Ala-Ala-Phe-p-nitroanilin, respectively) that could be fully blocked with single and dual 

peptidase inhibitors. Importantly, degradation of exogenously applied Met-enkephalin 

and Leu-enkephalin, but not of ß-endorphin and dynorphin, was prevented by 

thiorphan + bestatin or by RB3008. Conclusions. We show for the first time that 

degradation of opioids can be prevented by blockade of APN and NEP expressed on 

immune cells in painful inflammation. Enkephalins appear as preferred substrates for 

NEP and APN. Inhibiting the enzymatic degradation of opioids in peripheral inflamed 

tissue offers a promising strategy for the pain control without adverse side effects. 

Finally, the use of dual inhibitors has the additional advantage of avoiding different 

pharmacokinetics and bioavailability when two “single” inhibitors are employed. 

Poster Abstracts 68


P22 TRPC5 channel is activated by osmotically induced membrane stretch 

Sergio Soriano, Carlos Belmonte, Félix Viana, Ana Gomis 

Instituto de Neurociencias de Alicante. UMH-CSIC. Av. Ramón y Cajal s/n. 03550 

Sant Joan d'Alacant. Alicante. Spain 

Mammalian canonical transient receptor potential (TRPC) genes encode a family of 

nonselective cation channels that are activated following stimulation of G-proteincoupled 

membrane receptors linked to phospholipase C. TRPCs are molecular 

candidates for Ca2+ entry channels in brain since they are highly expressed in the 

nervous system, form Ca2+-permeable channels in recombinant expression systems 

and are activated by agonist that induce intracellular Ca2+ release. However, their 

mechanisms of activation and endogenous regulators remain poorly understood. We 

have been focused our studies on the TRPC5 channel. We used a combination of 

cellular Ca2+ imaging and patch-clamp techniques. We show that hypoosmotic stimuli 

(210mOsm), that cause membrane stretch, produce an increase in the intracellular 

calcium concentration in HEK293 cells transfected with TRPC5. During whole-cell 

recordings, the application of hypotonic stimulus activates the characteristic doubly 

rectifying currents mediated by TRPC5. The activation of TRPC5 is promoted with the 

co-expression of the histamine receptor type I (H1). Activation of inositol 

polyphosphate 5-phosphatase IV (5ptase IV), which reduce membrane 

phosphatidylinositol 4,5-bisphosphate (PIP2) levels, abolished hipotonically-evoked 

activation of TRPC5. This response was recovered by increasing intracellular PIP2 

levels through the patch pipette. We also show the expression of TRPC5 in sensory 

neurons suggesting that the activation of TRPC5 by osmotically induced stretch may 

play a role in sensory transduction. 

69 Poster Abstracts


P23 Opioid withdrawal increases TRPV1 activity in a PKA dependent manner 

Viola Spahn, Christian Zoellner 

Charite, Klinik fuer Anaesthesiologie und operative Intensivmedizin 

Vanilloid receptor type 1 (TRPV1) is a ligand-gated ion channel expressed on sensory 

nerves that responds to noxious heat, protons, and chemical stimuli such as 

capsaicin. TRPV1 plays a critical role in the development of tissue injury, inflammation 

or nerve lesions. Opioids such as morphine have been used widely for the treatment 

of many types of acute and chronic pain. Application of morphine leads to a 

dissociation of G-proteins and causes a reduced activity of adenylylcyclases (AC), 

resulting in a lower amount of cAMP. However, opioid withdrawal following chronic 

activation of the ƒÊ opioid receptor induces AC superactivation and subsequently an 

increase in cAMP and Protein Kinase A (PKA) activity. In the current project we 

investigated wether an increase in cAMP during opioid withdrawal increases the 

activity of TRPV1. In whole cell patch clamp and calcium imaging experiments opioids 

significantly increase capsaicin induced TRPV1 activity in a nalaxone and pertussis 

toxin sensitive manner. The role of different PKA phosphorylation sites at TRPV1 was 

investigated using site-directed mutagenesis. In summary, our results demonstrate 

that opioid withdrawal can increase the activity of TRPV1. These observations show a 

new mechanism underlying hyperalgesia during opioid withdrawal. 

Poster Abstracts 70


P24 A molecular dissection of TRPV1 sensitisation using the naked mole-rat 

Ewan St. John Smith, Gireesh Anirudhan, Gary Lewin 

Max-Delbrueck Centrum, Berlin-Buch, Germany 

Tissue inflammation leads to pain and behavioural sensitisation to thermal and 

mechanical stimuli called hyperalgesia. However, the African naked mole-rat (NMR, 

Heterocephalus glaber) has total behavioural insensitivity to capsaicin and acid. 

Additionally, nerve growth factor (NGF) fails to induce thermal hyperalgesia, a 

phenomenon dependent upon TRPV1. We aimed to discover why NGF fails to induce 

thermal hyperalgesia in NMRs. To investigate if NGF sensitises NMR TRPV1, wholecell 

patch clamp recordings were made from isolated DRG neurones. In mouse the 

isolectin B4 (IB4) labels DRG neurones that lack the NGF receptor TrkA and 

immunocytochemistry demonstrated the same pattern in NMR DRG cells. NGF 

sensitised the capsaicin-gated current in mouse DRG neurones that were IB4-ve (n = 

11), but failed to do so in either IB4+ve or IB-ve NMR DRG neurons (n = 14). NMR 

TRPV1 was subsequently cloned and found to possess a tyrosine at position 200 that 

in other species is fundamental for NGF-induced sensitisation in TRPV1. Furthermore, 

when expressed in CHO cells cloned NMR TRPV1 is activated by heat, capsaicin, pH 

and voltage in a similar fashion to TRPV1s from other species. We introduced NMR 

TRPV1 into DRG neurones from TRPV1-/- mice and in this context NGF caused 

potentiation of capsaicin gated currents in IB4-ve neurones (n = 8), but not in IB4+ve 

neurones (n = 3). This data suggests that there is something about NMR neurones or 

NMR TrkA that is responsible for the lack of sensitisation observed in NMR DRG 

neurones. To determine if the cellular environment is critical for NGF induced 

sensitisation, a new fibroblast cell line was established from NMR kidney tissue. Four 

clones were isolated and have reached population doublings in excess of 50, a formal 

criterion that indicates that these cell clones have been immortalized. We are using 

these cells to test the hypothesis of whether the cellular environment of NMR cells is 

responsible for the lack of NGF induced sensitisation. In summary we find that NGF 

fails to sensitise the NMR TRPV1 ion channel. This finding may account for the lack of 

NGF-induced thermal hyperalgesia in NMRs. Evidence suggests that there is a 

difference in NMR TrkA or downstream signalling that underlies the lack of 

sensitisation and these mechanisms are under investigation. 

71 Poster Abstracts


P25 Analysis of phosphorylation targets of cGMP-dependent kinase I involved 

in sensory axon bifurcation 

Agne Stonkute, Hannes Schmidt, Fritz G. Rathjen 

Max Delbrück Center for Molecular Medicine, Developmental Neurobiology Group, 


Our lab identified a cGMP signaling pathway via Npr2 (natriuretic peptide receptor 2) 

and cGMP-dependent protein kinase I (cGKI) that is important for axonal pathfinding 

and connectivity of sensory neurons. Upon arrival at the dorsal root entry zone 

(DREZ) of the spinal cord sensory axons bifurcate and grow further in both rostral and 

caudal directions. However, sensory axons of mice lacking cGKI do not bifurcate, they 

turn instead and grow in either caudal or rostral direction. This axonal pathfinding error 

has physiological implications. Electrophysiological studies showed that the absence 

of bifurcation leads to a loss of functional connectivity between sensory neurons and 

second order neurons within the dorsal horn. These initial studies raised the question 

which phosphorylation targets of cGKI mediate the process of axon bifurcation. In our 

search for downstream components of the cGMP signaling cascade in sensory 

neurons we analysed mice deficient for established phosphorylation substrates of 

cGKI. As a result we can exclude the Ena/Vasp family members Mena (mammalian 

Ena) and VASP (vasodilator-stimulated phosphoprotein) as candidate-proteins that 

could play a role in axon bifurcation. Furthermore, we screened for novel 

phosphorylation targets of cGKI using an antibody recognizing a phosphorylation 

consensus motif of cGKI. Western blots with the phospho-sequence specific antibody 

revealed several proteins that become phosphorylated upon stimulation of cGKI in 

dorsal root ganglia (DRG) or in the DRG-derived cell line F11. Currently we focus on 

the enrichment of these components to enable mass spectrometry analysis. 

Poster Abstracts 72


P26 New In Vitro mechano-assay reveals distinct intrinsic mechanosensitive 

subtypes in cultured sensory neurons 

Dmitry Usoskin, Per Uhlen, Patrik Ernfors 

Mol.Neurobiology /MBB, Karolinska Institute, Stockholm, Sweden 

We developed a novel in vitro assay for studying intrinsic mechanosensitive properties 

of sensory neurons. Dissociated sensory neurons are plated in specially developed 

compartmentized chamber. After several days in culture axons grow to another 

compartment where they are massively mechanically stimulated in a controlled way. 

Cell response is monitored using alive Ca2+ imaging of somas. Several functional 

subtypes could be reproducibly distinguished based on response profile in this assay. 

Some of subtypes have correlation with expressed neuronal marker, soma diameter 

and responsiveness to capsaicin. 

73 Poster Abstracts

P27 Function of Cav3.2 in Sensory neuron mechanosensitivity 

Rui Wang, Gary R. Lewin 


Molecular physiology and somatic sensation Group, Department of Neuroscience, 

Max-Delbrück-Center for Molecular Medicine Berlin-Buch, Germany 

Aim of investigation: Voltage-gated Ca2+channels mediate calcium influx in response 

to membrane depolarization and regulate intracellular processes such as contraction, 

secretion, neurotransmission, and gene expression. T-type Ca2+currents are 

activated by weak depolarization and are transient. They were first described in 

sensory neurons and also expressed in a wide variety of other cell types, where they 

were involved in shaping the action potential and controlling repetitive firing behavior. 

Previous findings from our lab demonstrated that the cDNA encoding the T-type 

Cav3.2 calcium channel was almost exclusively expressed by D-hair receptors in the 

DRG (Shin et al., 2003). D-hair receptors are very sensitive mechanoreceptors and we 

provided pharmacological evidence that the T-type current is required for normal 

excitability of this mechanoreceptor. In contrast several other groups have proposed 

that Cav3.2 is expressed by nociceptive neurons and that it may play a role in acute 

and chronic pain. In the present study we would like to determine whether Cav3.2 

channels contribute to the normal function of identified mechanoreceptors and 

nociceptors. Methods: In the current study we used Cav3.2 null mutant mice and 

tested the physiological properties of cutaneous sensory neurons with the in vitro skin 

nerve preparation. Results: Our results showed that mechanical sensitivity and 

thresholds of nociceptors (AM and C-fibers) were not changed in Cav3.2 null mutant 

mice compared to wild type. Among the mechanoreceptors the physiological 

properties of slowly adapting receptors (SA) were also not dramatically changed in 

Cav3.2 mutant mice. We noted a small increase in the mechanical latency of rapidly 

adapting mechanoreceptors (RA), mechanical latency is a measure of the threshold 

for activation. D-hair receptors have less mechanical sensitivity and higher mechanical 

thresholds for activation in Cav3.2 mutant mice. Conclusion: Our results demonstrate 

that Cav3.2 channels are directly or indirectly involved in setting the 

mechanosensitivity of mechanoreceptors but do not play an appreciable role in 

nociceptive sensory neurons. Acknowledgement: Supported by the DFG 

Poster Abstracts 74


List of Participants 

Sebastian Auer 


13125 Berlin 

Germany 

Fon: 030-9406-3415 

auer.sebastian@mdc-berlin.de 


Av. Ramón y Cajal, s/n 

03550 Sant Joan d´Alacant, Alicante 

Spain 

Fon: +34 965919530 

Fax: +34 965919549 

carlos.belmonte@umh.es and 

in.angelines@umh.es 

Carmen Birchmeier 

Robert Roessle Str. 10 

13125 Berlin 

Germany 

Fon: 030 9406 2403 

cbirch@mdc-berlin.de 

Steeve Bourane 

80 rue Augustin Fliche -BP74103 

34091 Montpellier 

France 

Fon: (33) (0)4 99 63 60 50 

Fax: (33) (0)4 99 63 60 20 

s.bourane@wanadoo.fr 

Michael Brecht 

Philippstr. 13 Haus 6 

10115 Berlin 

Germany 

Fon: 030 20936772 

Fax: 030 20936771 

michael.brecht@bccn-berlin.de 

Dominique Bröhl 

Robert-Rössle-Strasse 10 

13125 Berlin 

Germany 

Fon: +49-30-9406 3359 

d.broehl@mdc-berlin.de 

75 List of Participants 

Isabelle Bachy 

Sheelesvag 1 

17177 Stockholm 

Sweden 

Fon: +46 8 524 87866 

isabelle.bachy@ki.se 

Yinth Andrea Bernal Sierra 

Robert- Rössle Straße 10 

13125 Berlin 

Germany 

Fon: +493094063789 

yinth-andrea.bernal-sierra@mdcberlin.de 

Walter Birchmeier 


13125 Berlin 

Germany 

Fon: 030 9406 3278 

Fax: 030 949 7008 

wiss.vorstand@mdc-berlin.de 

Janko Brand 

Torstr.104 

10119 Berlin 

Germany 

Fon: +49 30 9406 3275 

Janko.brand@mdc-berlin.de 

Christian Brenneis 

Theodor Stern Kai 7 

60590 Frankfurt 

Germany 

Fon: 0049069630183103 

Fax: 0049069630183378 

C.Brenneis@med.uni-frankfurt.de 

Austin Brown 

Beckman 115, 279 Campus Dr. 

94305 Stanford, Ca 

USA 

Fon: 650-723-8580 

austin.brown@stanford.edu

Jean-François Brunet 

46 rue d'Ulm 

75005 Paris 

France 

Fon: 01 44 32 23 21 

Fax: 01 44 32 23 23 

jfbrunet@biologie.ens.fr 

Ombretta Caspani 

Hindenburgdamm 30 

D-12200 Berlin 

Germany 

Fon: 030 84452131 

ombretta.caspani@charite.de 

Marta Ceko 

Robert-Rössle Str 10 

D 13122 Berlin-Buch 

Germany 

Fon: 0049 1577 198 9404 

marta.ceko@gmail.com 

Justyna Cholewa-Waclaw 

Robert-Rössle-str. 10 

13092 Berlin 

Germany 

Fon: +49 30 9406 3772 

jcholewa@mdc-berlin.de 

Thomas Delanne 

1 avenue Eugene Schueller 

93600 Aulnay sous Bois 

France 

Fon: 33 1 58 31 71 38 

Fax: 33 1 47 56 42 58 

tdelanne@rd.loreal.com 

Usoskin Dmitry 

Scheeles väg 1, A1 

171 77 Stockholm 

Sweden 

Fon: +46-8-52482269 

Fax: +46-8-34 19 60 

dmitry.usoskin@ki.se 



80, rue Augustin Fliche, 

34091 Montpellier cedex 5 

France 

Fon: (33) 4 99636041 

Fax: (33) 4 99636020 

carroll@univ-montp2.fr 

Cecile Cayla 


12200 Berlin 

Germany 

Fon: 01636927046 

cecile.cayla@charite.de 

Li-Yang Chiang 

Robert-Rössle-str. 

13125 Berlin 

Germany 

Fon: +49-30-94063783 

Fax: +49-30-94062793 

li-yang.chiang@mdc-berlin.de 

Elvira De la Peña García 

Campus de San Juan 

03550 San Juan de Alicante. Alicante 

Spain 

Fon: +34965919212 

Fax: +34965919561 

elvirap@umh.es 

Bristol Denlinger 

Avda. Ramón y Cajal s/n 

03550 San Juan de Alicante 

Spain 

Fon: +34965919344 

Fax: +34965919561 

blayne@umh.es 


Scheelesv 1 

S-17177 Stockholm 

Sweden 

Fon: +46 70 3297659 

Fax: +46 8 341960 

patrik.ernfors@ki.se 

List of Participants 76


Axel Fischer 

Augustenburger Platz 1 

13353 Berlin 

Germany 

Fon: +49-30-450 559851 

Fax: +49-30-450 559851 

axel.fischer@charite.de 


Rpbert-Rössle-Strasse 10 

13125 Berlin 

Germany 

Fon: +493094063785 

agarratt@mdc-berlin.de 

Ana Gomis 

Av. Santiago Ramón y Cajal 

03550 Sant Joan d'Alacant. Alicante 

Spain 

Fon: +34 965919597 

Fax: +34 9659549 

agomis@umh.es 

Martyn Goulding 

10010 North Torrey Pines Rd 

CA 92037 La Jolla 

USA 

Fon: +1858 4534100 x1558 

goulding@salk.edu 

Sven Hendrix 

Charitepl. 1 

10117 Berlin 

Deutschland 

Fon: 0176 411 93 272 

sven.hendrix@charite.de 

Jing Hu 

Robert-Rössle-str. 

13125 Berlin 

Germany 

Fon: +49-30-94063783 

Fax: +49-30-94062793 

jinghu@mdc-berlin.de 


Henning Frenzel 

Weichselstrasse 24a 

10247 Berlin 

Germany 

Fon: +491793936630 

henning.frenzel@mdc-berlin.de 

David Ginty 

725 N. Wolfe Street, PCTB 1015 

21205 Baltimore, Maryland 

USA 

Fon: 410-614-9494 

Fax: 410-614-8423 

dginty@jhmi.edu 


279 Campus Dr 

94305 Stanford, CA 

USA 

Fon: 650-721-5976 

Fax: 650-725-8021 

mbgoodman@stanford.edu 

Dagmar Hackel 

Krahmerstr 6-10 /FEM 

12207 Berlin 

Germany 

Fon: 030-84453867 

dagmar.hackel@charite.de 

Eric Honore 

660, route des Lucioles 

06560 Valbonne 

France 

Fon: 33 4 93 95 77 45 

Fax: 33 4 93 95 77 04 

honore@ipmc.cnrs.fr 

Tim Hucho 

ihnestrasse 

14195 Berlin 

Germany 

Fon: 030-8413 1243 

Fax: 030-8413 1383 

hucho@molgen.mpg.de

Ines Ibanez-Tallon 

Robert Rossle 10 

10125 Berlin 

Germany 

Fon: +49-30-9406-3411 

Fax: +49-30-9406-2430 

ibanezi@mdc-berlin.de 

Anita John 

Albert-Einstein-Allee 11 

89081 Ulm 

Germany 

Fon: 0551-397007 

ajohn@uni-goettingen.de 

Alexey Kozlenkov 

Robert-Roessle str. 10 

13125 Berlin 

Germany 

Fon: +49 30 9406 3212 

alexey.kozlenkov@mdc-berlin.de 

Dominika Labuz 


12200 Berlin 

Germany 

Fon: +49 30 84453867 

dominika.labuz@charite.de 

Michaela Langer 

Robert Rössle Str. 10 

13125 Berlin 

Germany 

Fon: +49 30 9406 3720 

Fax: +49 30 9406 2206 

langer@mdc-berlin.de 

Stefan Lechner 

Robert Rössle Str. 10 

13125 Berlin 

Germany 

Fon: 030-94062853 

stefan.lechner@mdc-berlin.de 


Julia Jira 


12200 Berlin 

Germany 

Fon: 03084452131 

julia.jira@charite.de 

Jane Johnson 

5323 Harry Hines Blvd 

75390-9111 Dallas 

USA 

Fon: 214-648-1870 

Fax: 214-648-1801 

jane.johnson@utsouthwestern.edu 

Rohini Kuner 

Im Neuenheimer Feld 366 

69120 Heidelberg 

Germany 

Fon: 06221-548289 

Fax: 06221-548549 

rohini.kuner@pharma.uni-heidelberg.de 

Francois Lallemend 

Scheeles väg 1-A1-plan2 

171 77 Stockholm 

Sweden 

Fon: +46 8524 876 20 

francois.lallemend@ki.se 

Liudmila Lapatsina 

Robert-Roessle str 10 

13125 Berlin 

Germany 

Fon: +49-30-9406-3783 

l.lapatsina@mdc-berlin.de 

Gary Lewin 

Robert-Rössle str 10 

13192 Berlin 

Gemany 

Fon: 030 94062430 

glewin@mdc-berlin.de 




660 Route des Lucioles 

06560 Valbonne - Sophia Antipolis 

France 

Fon: 33 (0)4 93 95 77 20 

Fax: 33 (0)4 93 95 77 04 

lingueglia@ipmc.cnrs.fr 

Halina Machelska 

Krahmerstr. 6 

12207 Berlin 

Germany 

Fon: +49-30-8445 3851 

halina.machelska@charite.de 


Wolfson Wing, Hodgkin Building, Guy's 

Campus 

SE1 1UL London 

UK 

Fon: +44 207 8486270 

Fax: +44 207 848 6165 

stephen.mcmahon@kcl.ac.uk 

Lorne Mendell 

Life Science Building 

NY 11794 Stony Brook 

USA 

Fon: 631 632 8632 

lorne.mendell@sunysb.edu 

Goodman Miriam 

B-111 Beckman Center, 279 Campus Dr 

94305 Stanford, CA 

USA 

Fon: 650-723-3100 

Fax: 650-725-8021 

mbgoodman@stanford.edu 

Thomas Mueller 

Robert-Roessle-Str. 10 

13125 Berlin 

Germany 

Fon: 49 30 9406 2842/3359 

Fax: 49 30 9406 3449 

thomu@mdc-berlin.de 


Qiufu Ma 

1 Jimmy Fund Way 

MA 02115 Boston 

the United States 

Fon: 617 632-4594 

Qiufu_Ma@dfci.harvard.edu 

Sören Markworth 


13125 Berlin 

Germany 

Fon: +49-(0)30-9406-2853 

soeren.markworth@mdc-berlin.de 

Peter McNaughton 

Tennis Court Rd 

CB1 2ET Cambridge 

UK 

Fon: +44 1223 334012 

pam42@cam.ac.uk 

Nevena Milenkovic 

Robert Rossle Str. 10 

13092 Berlin 

Germany 

Fon: +493064063783 

milenkovic@mdc-berlin.de 

Rabih Moshourab 

hindenburgdamm 30 

12200 berlin 

germany 

Fon: 84452733 

rabih.moshourab@charite.de 

Robert Naumann 

Schlegelstr. 5 

10115 Berlin 

Germany 

Fon: 030 21230778 

robert.naumann@bccn-berlin.de

Dinah Nockemann 

Krahmerstr. 6-10 

12207 Berlin 

Germany 

Fon: 030-84452131 

dinah.nockemann@charite.de 

Thomas Park 

840 West Taylor St. 

60607 Chicago 

USA 

Fon: 312 413 3020 

Fax: 312 996 2805 

TPark@uic.edu 

Kate Poole 

Robert-Rössle Str 10 

13125 Berlin-Buch 

Germany 

Fon: (030)94063789 

kate.poole@mdc-berlin.de 

Michael Schäfer 


12200 Berlin 

Germany 

Fon: 030 8445 2731 

micha.schaefer@charite.de 

Robert F. Schmidt 

Röntgenring 9 

97070 Würzburg 

Germany 

Fon: 0931 31 2639 

Fax: 0931 592 10 

rfs@mail.uni-wuerzburg.de 

Mikael Schnizler 

Pauwelsstraße 30 

52074 Aachen 

Germany 

Fon: 0241-80888-06 

mschnizler@physiology.rwth-aachen.de 


Otilia Obreja 

Theodor-Kutzer-Ufer 1-3 

68167 Mannheim 

Germany 

Fon: +49 621 3835619 

Fax: +49 621 3831463 

otilia.obreja@anaes.ma.uniheidelberg.de 

Andrés Parra 

ctra N-332 km 37 

03550 Alicante 

Spain 

Fon: +34 965919344 

aparra@umh.es 

Fritz G. Rathjen 

Robert-Rössle-Str.10 

13092 Berlin 

Germany 

Fon: 49 30 9406 3709 

Rathjen@mdc-berlin.de 

Hannes Schmidt 


13125 Berlin 

FR Germany 

Fon: +49-30-94063583 

hannes.schmidt@mdc-berlin.de 

Yvonne Schmidt 


12203 Berlin 

Germany 

Fon: 030 8445 2131 

yvonne.schmidt@charite.de 

Anja Schreiter 


12203 Berlin 

Germany 

Fon: 0049 30 84453867 

anja.schreiter@charite.de 



Jan Siemens 

600 16th Street 

94158-2517 San Francisco 

USA 

Fon: +415-476-0432 

Fax: +415-502-8644 

jan@cmp.ucsf.edu 

Viola Spahn 

Krahmerstr. 6-10 

12207 Berlin 

Germany 

Fon: 030-8445 2131 

Fax: 030-8339 389 

viola.spahn@charite.de 

Agne Stonkute 

Robert-Roesslestr. 10 

13092 Berlin 

Germany 

Fon: 030 94063316 

stonkute@mdc.berlin.de 

Annika Stuerzebecher 

Robert Rössle Straße 10 

13125 Berlin 

Germany 

Fon: 030/94063415 

annika.stuerzebecher@mdc-berlin.de 

Jean Valmier 

80 augustin Fliche 

34090 Montpellier 

France 

Fon: 0033499636007 

Fax: 0033499636020 

jean.valmier@univ-montp2.fr 

Hong Wang 


12200 Berlin 

Germany 

Fon: 00493084452131 

hong.wang@charite.de 

Dominik Wiemuth 

Pauwelsstraße 30 

52074 Aachen 

Germany 

Fon: 0241-8088814 

dominik.wiemuth@uni-wuerzbug.de 


Anantha Krishna Sivasubramaniam 

coppistr. 14 

10365 Berlin 

Germany 

Fon: 015775769488 

anand987@yahoo.com 

Ewan St. John Smith 

Robert Rössle Strasse 10 

13125 Berlin 

Deutschland 

Fon: 00493094063783 

ewan.smith@mdc-berlin.de 

Robert Storm 


13125 Berlin 

Germany 

Fon: +49 - 30/9406 2842 

rstorm@mdc-berlin.de 

Andrew Todd 

University Avenue 

G12 8QQ Glasgow 

UK 

Fon: 0141 330 5868 

A.Todd@bio.gla.ac.uk 

Daniel Vardeh 

Ladenburgerstr. 19 

60120 Heidelberg 

Germany 

Fon: 06221 9143676 

danv@gmx.de 

Rui Wang 

Robert-Rössle-Str 10 

13092 Berlin 

Germany 

Fon: 03094062853 

ruiwang@mdc-berlin.de 

John Wood 

GowertStreet 

wc1e 6bt London 

UK 

Fon: 02076797800 

j.wood@ucl.ac.uk

Clifford Woolf 

149 13th Street 

02129 Charlestown 

USA 

Fon: 617 724 3622 

cwoolf@partners.org 

Sandra Zurborg 

Krahmerstraße 6-10 

12207 Berlin 

Germany 

Fon: 03084452131 

sandra.zurborg@charite.de 


Hanns Ulrich Zeilhofer 

Winterthurerstrasse 190 

CH-8057 Zurich 

Switzerland 

Fon: -41-44-6355912 

Fax: -41-44-6355988 

zeilhofer@pharma.uzh.ch 



Author Index 

A D 

Alvania, Rebecca S. T6 de la Peña, Elvira P9 

Angioni, Carlo P2 Dina, Olayinka T17 

Anirudhan, Gireesh P24 

Anjum, Farzana T19 E 

Auer, Sebastian P1 

Erdmann, Bettina T10 

B P7 

P14 

Bader, Michael P6 Ernfors, Patrik T2 

Basbaum, Allan I. T21 P26 

Bautista, Diana M. T21 

Belmonte, Carlos T30 F 

P9 

P22 Feil, Robert T5 

Birchmeier, Carmen T4 Frahm, Christina T10 

T10 Frahm, Silke P18 

T20 Frenzel, Henning P17 

P3 

P8 G 

P16 

Bormuth, Ingo P3 Garratt, Alistair T10 

Boyce, V. T12 P10 

Brack, Alexander T24 Geisslinger, Gerd P2 

P12 Ginty, David D. T6 

Brecht, Michael T19 Glazer, Josh T21 

Brenneis, Christian P2 Gomis, Ana P22 

Britsch, Stefan P16 Goodman, Miriam T7 

Bröhl, Domenique T20 P4 

P3 Gore, Carmen P21 

Brown, Austin P4 Goswami, Chandan P11 

Brunet, Jean-François T28 Griffel, Carola T10 

Brylka, Heike P16 

Buttgereit, Jens T5 H 

C Hackel, Dagmar P12 

Hendrix, Sven P13 

Carroll, Patrick T3 

Caspani, Ombretta T8 Heppenstall, Paul T8 

P5 P5 

Cayla, Cecile P6 P15 

Chiang, Li-Yang P7 P20 

P14 Hofmann, Franz T5 

Cholewa-Waclaw, Justyna P8 Honore, Eric T9 

Copeland, Neal G. P16 Hori, Kei P3 

Coste, Ovidiu P2 Hu, Jing T22 

Crenshaw III, E. Bryan P16 P7 

P14 

83 Author Index


Hucho, Tim T17 

P11 

M 

Ma, Qiufu T29 

I Machelska, Halina T24 

P20 

Ibanez-Tallon, Ines T22 P21 

P1 Madrid, Rodolfo P9 

P18 Mandai, Kenji T6 

Markworth, Soeren P18 

J Martarelli, Benedetta P8 

Maurer, Marcus P13 

Jenkins, Nancy A. P16 McMahon, Stephen T31 

Jira, Julia P15 McNaughton, Peter T11 

John, Anita P16 Mendell, L. M. T12 

Johnson, Jane T16 Milenkovic, Nevena T10 

Jordan, Jens P18 Mousa, Shaaban A. P20 

Jordt, Sven-E. T21 P21 

Julius, David T21 Müller, Thomas T4 

Jüttner, Rene T5 T20 

K 

P1 

P16 

N 

P3 

Nave, Klaus-Armin P3 

Koch, Manuel P7 

Kuhn, Julia T17 P 

Kuner, Rohini T14 

P. A. Heppenstall, D. P19 

Nockemann, C. Stein 

L Park, Thomas T18 

Peters, Eva M. P13 

Labuz, Dominika T24 Petruska, J. T12 

P6 Picker, Björn P13 

P12 

Lechner, Stefan G. P17 R 

Levine, Jon T17 

Lewin, Gary T10 Ramanan, Narendrakumar T6 

T18 Raouf, Ramin T27 

T22 Rathjen, Fritz G. T5 

T26 P25 

P7 Rittner, Heike T24 

P14 P12 

P17 Roques, Bernard P. P21 

P18 Rugiero, Francois T27 

P24 

P27 S 

Liao, Zhiwen P4 

Lingueglia, Eric T23 Schäfer, Michael P6 

Liu, Pentao P16 Schäffer, Susanne T5 

Schmidt, Hannes T5 

P25 

Author Index 84


Schmidt, Yvonne T24 Y 

P20 

Scholich, Klaus P2 Yurgionas, Brian T8 

Schreiter, Anja T24 

P21 Z 

Siebenhaar, Frank P13 

Sieber, Martin T4 Zeilhofer, Hanns Ulrich T25 

Siemens, Jan T21 Zhang, Xuming T11 

Soriano, Sergio P22 Zoellner, Christian P23 

Spahn, Viola P23 Zurborg, Sandra T8 

St. John Smith, Ewan P24 P5 

Stein, Christoph P6 

P21 

Stonkute, Agne T5 

P25 

Storm, Robert T4 

Strehle, Michael P3 

Stucky, Cheryl L. T21 

Stuerzebecher, Annika T22 

T 

Tansy , Donovan-Rodriguez P9 

Todd, Andrew T13 

Treier, Mathias T20 

Tsuruda, Pamela R. T21 

U 

Uhlen, Per P26 

Usoskin, Dmitry P26 

V 

Viana, Felix P9 

P22 

W 

Wang, Rui P17 

P27 

Wende, Hagen P3 

Wickramasinghe, S. Rasika T6 

Wildner, Hendrik T20 

P8 

Wood, John T27 

Woolf, Clifford J. T1 

85 Author Index

conference booklet - MDC

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