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Neural Progenitor Cells in the
Huntington's Disease Hurnan Brain
Maurice A. Curtis
A thesis submitted in fulfilment of the requirements for the Degree of
Doctor of Philosophy in Anatomy and Pharmacology at The University of
Auckland, May, 2004
THE UNIVERSITY OF AUCKLAND

" ... tkefunafional spec-ializaiton ef the brain imposes anthe neurones two great lacunoc;
p'rolifetation inqbility and irreversabitity af intrapratoplasmie fficrentiation It is for
this reaaan tkat, iQn:ee the development was ended, the faants af grwth and regeneration
af wcoas and derdriNes dried up irreuocab,Iy. In adult eenters tke nente paths are
somahingfi'Ned, ended, imrnutabrc-. Everything may dle, nothing may he regenerated. lt'h
for the science of thefutwre ta e,hange, if possible, this harsh d,ecree.,,
Santiago,Rarnon Y. Cajal (lgl3)
Nobel prize in Medicine IgA6
(Ramon y Cajal, 1991)

The recent demonstration of endogenous progenitor cells in the adult mammalian brain
raises the excitirrg possibility that these unclifferentiated cells may be able to generare
new neurons fbr cell replacement in diseases such as Huntington's disease (HD).
hevious studies have shown that neural stem cells in the rodent brain subependymal
layer (SEL), adjacent to the caudate nucleus. proliferate and differentiate into neurons
and glial cells but no previous study has characterised the human SEL or shown
neurogenesis in the diseased human brain. In this study. histochemical and
immunohistochemical techniques were used to demonstrate the regional anatomy and
staining characteristics of the normal and HD brain SEL using light and laser scanning
confocal microscopy. The results demonstrated that the normal and HD SEL contained
migrating neuroblasts, glial cells and precursor cells but there were more of each cell type
preseltt in the HD brain, and that the increase in cell numbers correlated with HD
neuropathological grade. The nonnal and HD SEL was stained with a proliferative
marker, proliferating cell nuclear antigen (PCNA), to label divicling cells. The results
showed a significant increase in the number of dividing cells in the HD brain that
correlated with HD grade and with CAG repeat length. Furthermore, the results showed
that neurogenesis had occurred in the SEL as evidenced by co-localisation of pCNA and
the neuronal marker Blll-tubulin. Also. gliogenesis had occumecl in the SEL as evidenced
by the co-localisation of PCNA with the glial marker GFAP. These studies also revea.led
a 2'6 fold increase in the number of new neurons in the HD SEL. PCNA positive cells
were distributed throughout the SEL overlying the caudate nucleus but most notably the
ventral and central regions of the SEL adjacent to the cauclate nucleus contained the
highest number of prolif'erating cells. I examined the SEL fbr mature cell markers and
demonstrated many of the same cell types that are present in the normal striaturn. With
the exception of neuropepticle Y (NPY) neurons, there was a reduction in the number of
mature neurons in the HD SEL. The NPY neurons were more abundant in the HD SEL
suggesting they play a role in progenitor cell proliferation. The results in this thesis
provide evidence of increased progenitor cell proliferation and neurogenesis in the
diseased adult human brain and indicate the regenerative potential of the human brain.
These findings may be of rnajor relevance to the development of therapeutic approaches
in the treatment of neurodegenerative diseases.
llt

ACKNOWLEDGEMENTS
I wish to specifically thank the members of the Anatomy department and a number of
other people who have directly ancl indirectly influenced this phD work throughout the
past 3 years.
First and foremost I would like to thank rny principal supervisor hofessor Richard Faull.
My undergraduate degree was not the prerequisite required to do a Masters degree in
Richard's subject area but Richard successfully took my case before the appropriate
committees and took me on as a Masters student and subsequently as a phD student.
Richard has always been enthusiastic and positive throughout the highs and the lows and
has inspired and mentorecl me in every aspect of this thesis. I will be forever grateful for
the chance you took on me which has changed ancl improved my rife.
I would like to thank Professor Mike Dragunow who, as my co-supervisor. has provided
me with new ideas and fresh ways of looking at olcl problems. Mike has such an
infectious enthusiasm for science that it is virtually impossible to leave Mike's office
without being so fired up that you want to cancel your holidays and spend the time doing
exciting experiments. Thank you for ail your ideas and enthusiasm.
Also, I would like to thank Dr Bronwen Connor (co-supervisor). Bronwen has always
been prompt with her fee

epresented in chapter 5 and has weathered some of my fairly inclement questions
regarding statistics. Thank you for all your assistance. Thanks also to Jocelyn Bullock for
your work cutting sections to obtain Weslern blotting tissues, discussion about interesting
destinations and for lending/giving me quality literature of the adventure variety.
While working toward this thesis I have had the privilige of being surrounded by some
exceptionally bright PhD students who have provided me with scientific ideas and
assistance, social interactions and friendship. I would like to thank the PhD students in
the Depertments of Anatomy and Pharrnacology, namely Rachel Cameron, Palingu
Dhanapala, Carissa Fonseca. Hannah Gibbons. Monica Kam, Andree Pearson, Suzanne
Reid and Doris Thu. Best wishes in your studies and for the future. A special thanks to
Rachel for the much needed. sanity-inducing hot chocolate breaks, proofreading. hearing
my crazy stories and for the listening ear.
To John Klistiansen, Daniel McGowan and Glenn Pennycook, thanks for all the climbing
adventures and CCMC trips to talk about it. You are top friends and have always offered
light relief, hard case conversation and friendship regardless of the situation. I have been
very fortunate to have some supportive church friends and in particular Andrew
Schwamm, Tim Matsis, Jeremy Dixon and Leah Howard. Thank you for your church
outreach, philosophical conversations and for coming sailing with me in gale force winds.
These words poorly articulate the importance your friendship is to me.
The two most supportive people in rny life have always been Ken and Lorraine Curtis
(my parents). They have provided me with unerring and immeasurable love and support.
They have always listened for hours on the telephone while I raved on about my
disasters, my successes and my plans for the future. Thank you both for all you have done
for me.
Finally, I wish to acknowledge and thank the Neurological Foundation of New Zealand
for their very generous financial support of me as a Miller scholar. Also, the Health
Research Council of New Zealand for their support in terms of working expenses and
equipment. I am very grateful for this support.

1. B. connor, w. van Roon-Mom. M.A. curtis, M. Dragunow,
R.L.M Faull (2001)
Stem Cells and Neurodegenerative Diseases
New Zealand Medical Journal. l14:477_479
2. M.A. Curtis, E.B. penney, A.G. pearson, W.M.C. van Roon-
Mom, N.J. Butterworth, M. Dragunow, B. connor, R.L.M. Faull
(2003)
Increased Cell Proliferation and Neurogenesis in the Adult Human
Huntington's Disease Brain
Proceedings of the National Academy of sciences of the united
States of America 100: gO23-gOZ7
3. M.A. Curtis, B. Connor, R.L.M. Faull (2003)
Neurogenesis in the Diseased Aclult Human Brain: New Therapeutic
Strategies for Neurodegenerati ve Diseases
Cell Cycle 2: 4ZB-430
vi

1r1l

2-'4.2Huntingtorr'sd.i'seascgadingseale..........|.r.'.!:r.]rt..ti.i
2'5 The striaturn: cell types and relatisnship to the subependy,mal trayer.-...,.-.,.-.....,.......33
2.6 Therap-e.utic approaches to Huntington' s disease,.,....
2.6.1 Neural transplaimtion..,........... r,..!...d......r,. ...-,_...........-..37
2.6.2 hogenitor eell rep1acement.,,.............. .........40
Chapten 3
ANATOMY OF TI{E SI]BEPENDYMAL LAER
IN TIffi NORIvIAL AND HL]-NTINGTOIN'S
DISEASE HUMAN BF|AIN... .....r...................:42
3.2ltIrtteriaIsandmethodS...'......,...".'.'.rn....i..it.i...i,..t.'l.!...r...
3.2.1 Flur,nan tissue co11ec6on".......... .....r......rr.....:..i...i..i.i:.:.;.i .,.-..,43
3,2.2 rissue proce$sins.......,........ ....--:....:.,::.."".":... ........4$5
3.2.3 Heaurotoxylin & essin arrd luxol fast lue:staininE....... ....,._,......,...4S
3.2,4 knage digitisarion
3.2.5 trnage anal ysi s...., ; d. i.... ii.!..,.,.
3.3.1 The anarom), of the SEL in the normal and
Huntinglon's disease brain..,.,,.... ._r....r.........,. .........,.,....4j
3.3.I.1Histological classification of the cells in the
adult human subependymal lay_e.r......
3,.3.l.?Iocalisation of the differenr cell tSpes in the
subepcnd;ymal layer........ r..{ !...... r. ;,.
vilt

3,3-2 comparison of the thickness of the subependymal layer in
uorural and Huntington's disease brains.._...,..,i,.bj.........!.,. ..................52
3.3.3 Quantification of cells in ttre subependyrnal layer in norural
and, Huntington's tlisease brains.............r,.:.....,.-.. ............i,,...i.,......,.5g
3.3,4 compnrison of the dif-ferent cell types present in the adulr
subependymal layer in normal and Huntington's disease brains...........,,............61
3.4.1 The general anatomy of the subependymal layer-,,,...
342 The Huntington's disease subependy,mal layeris thicler
than,that of the norrnal brain subependymat layer.............,......q...ii...._,,,.........65
3,4.3 The Huntington's disea$e bmin subependynlal layrer contains
mor€:cells than that of the normal subependyr,nal layer".,...,..,........:r....!.............66
3.4.4.ThEre are rnore type A, B and C eells in the
Huntin'gton's disease brain subependymal layer
compared with the normal brain subependymallayer..........................ri .......,,..67
3.6 Conclusion...........
CIrapter 4
ANATOMY OF THE SUBEPENDYMAL LAYER
IN THE NORMAL AND TITINTINGTON'S
D[sEAsEHI.'MANBRAIN....iil.};.'....j.;..i.......:..tt.....'.
4.2 Materials and rne_thods.....--....... ........_..72
4.2.1 Human tissue collection,. ......u........,......r1i...n..r.!- "...........72
lx

4.2 :2 Imrnanohistoaherni sty.... -.......
4.2.2.I Section preparation ...................,.;...........r... ......73
4.2.2Z ptirary antibodies,........_...i.,.r...,.;....,...;.i.r..r .i..._.!.....r.. .,.oT4
4.2,2,3 Secondary antibodiesr. .......r..,....r.r..................J4
4.2.2ATertiaryantibodies..j].......r....i;j.....i....t,i.....r...
4.2.3 Imrnunofluorgscence:i,..!-.............,r.,r..,r..;-..........i.,.i......-. ...................75
4.2.3.1Section Fr€Faratiom..............r..!,......ri.,...--. ......J..!..............25
4.2.3,2himury antibodies.. ..r.ij.,..i............75
4.2.3.3 Fluorochromqs..........,.
4.2.3.AIrnageanalysisandcellcounting.........'......
4.2.4 IVestern blotting..,......i.,....!-.rr..r........,!,...,..t..i..,....i..,r.,. ...................,7V
4.2'4..IProparationofgels..'.......'......'..l..i.
4.2,42 preparation of riunples. ..i!.r....i......!. ..........-".....7g
42,4.3lunrrunob-Iotting..............,..,r.,......j.i!.,.i...,!..,.......... .,.,............g0
4.3.il hogenitor cell proliferation in the hurnan norrnal and
Huntington's disease s_ubependymal 1ayer......,.,. ............g1
4.3.2 comparison,of the numberof proriferating ceils in the
normal and Huntingtoj's disease subependymal layer........,.j-.....*.-...r:.,r;r.ri;;.g2
4.3.3, Comparison of the pathologieal grade of Huntington's
disease with thenumber of proliferating cells in the
subependymal layer......
4.3 4 conrparissn of the number of cAG repeats in the rrr 5
gene in Huntington's direase with the number of Broliferating
cells in the subepe.ndymal |ayer........,.r..-..qi,.,....i;.;!..1............ .........g4
4.3.5 ,{nalysis qf the specificity of the PCNA antibody for
detecting cell proliferatiqn.................--r;.,......,.,;........i,..i ......-.........,..E5
4.3.6 Western blot analysis of the subependymal laye.r...... ,.!.....ij..i..i.!....-.g5

4.5
4.3.7'The lsealisation of proliferating, neuronal and glial
cell rnarkers in the subependymal |ayer......r.......r...:.;n..o.r..!:.......'.i.......',,............86
4.3.8 Neuroggnesis arid gliogenesis in the human subependymal layer......................90
4.3.9 The nrumber of PCNA/Bill=tuhulin double-labe-lled cells in the
normal and Huntington's disease subependym,al 1ayer.......... ".........93
4,4.I lncreased numbers of progenitor cells in the HD subependymal layer.....'..,.....97
4.4.2 Neurogenesis and gLi'ogenesis in the Huntington's disease
subependyrnal layer...... .i...i!....;..'r......,.........98
4.4,3 lncreased neurogenesis in the Hunti4gton's
diseases,ubependyrnalIa er.......... .......;..-...j.;....i;..,;...,.,......,.....'......101
xl

5.24 &ll counting,......:........,..r..........-...i.,...,.... j,...iij.i..!.. .,r.r..r:i.......,,......113
5.2.5Stuisticalanalysis................;!............'ji.i
5'3 Results ............ilT
5.3.1 Cornparison;of pCNA positive cell'umbers in:the
normal and Huntington's disease sub,ependymal layer......
5.3.2The numbEr of pCNA.positive cells in the subependyanal
layer in dorsal, nriddre and ventral areas of normar and
Huntington's disease braing.....-. q..j...ri.?i!...,....,....,.........1 I E
5.3.3 The number sf pcNA positive cells in the zubependral
layer in nostal, cenhar and caudar regions. of the nort'al and
Huntington' s disease brains........
5.3,4 The nurnber of FCNA positive ceils in the subependymal
layer in different roc.'atio''s in the nqrmar and Huntington,s
diileaee brains..,...." ...i..;....i.i...r.._..,.-...,......121
5.4.1 There is a variation in theregional distibution of pCNA
poniti]vecellsinfienorrrra|brainsubepend,yma|Iayetr.,.......'..
5.4.2 Ttnere is a vari.ation in the regional distribution sf pcNA
positive eells in the Huntington's disease brain
su'beperdfmalLlayer......... .,......It:s
5-4.3 There is a variation in rhe disfiburion of pcNApositive
cefls in different locati n of the subependymal layer...... ...........,.,126
xu

Chapten 6
CI,MMICAL ANATOVTY Or
THE SUBEPENDYI\{AL LAYER.....!....IIr r,. t...
6.1 Introduc
6"2 Mabrial
6,2-l Human bgste collection.. ...;i.a,q....i,.!i.!....!:....?.t... ....,,....Lg'z
6,?2Immunohistocherrristry.............---.b.r,.i..r...i..;i!,..,..._.....r
.........".........133
6.2.2.1 Section preparation
6.2.22 himary anribodies. .i..r........r.i.. ....,135
5,2.2,3 Secondary antibodies...i.,...;.rr.!..!... ...........!..i.j.r..",.,..-..........13d
6.2.2.4 Tefli4ry,arrtibodies..i. r.q.t,... r.i...
i5.2.3luu4e digitisation .,,.136
6.3.1 Calbinditr illrnunoreactivit5r in the subependyraal layer......!...........ii.ri.-.........13?
6.3.|.tN:grnral....'.!....-.'l.]...'r;..bi.r'.i.i.t'....'...t.
6.3. N .2 Elunfi ngton' s disease.,.
6'3.2 Enkephalin ininnunoreactivity in the subependyr,nar layer
6.3.2.L Ngnnal.,..,.tri...!.....r.......i..r...i..r.......,....{...rr-..,
-........-......139
6.3.2,,2 Huntington'g dieease... .r,....i.-.......ir...,.i....".,.,..,r,...,.........,."I39
63'3 substane-e P imhunoreastivity in the subepindymal layer
6.3.3.1 Nt0rmal,",,.. ...i,.!,.....,..i....!i!..ir!.....,... ........".....,.. t3g
6.3 .3.2 Huntingtoa' s disease,..
6.3.4 Parvalburnin irmunoreactivity in the subependynral layer
63,4.INormel.'.".'.l........J.a..-.|...iJrri.i|i;.|r..|.l
6 "3.4.2 Hun tington' s drsease...
xiii

6.3.5 eahetinin irnrnunoreactivity in the subeper'rdy-mal layer
6.3.5.1 Norural...... ........"........142
6.3.5.2Huntiugton's disease",...................,.......,.....r.....i..d..i...!ii.i..,.-. ..............L42
6.3.6 Choline-acetyl transferase immunoreactivity in the subependymal layer
6.3,6.lNormal......,:.i.i.i.t.',,........!.!r,.r.j..i.....r.
6.3.6,2 Huntington''s disease... .,....,.....i."...,...............,.143,
6.3.7 fl|.{ourqpsptide Y immur,aoreactivity in the subependymal layer
6.3,7,L Norrn'al........i........'......,.
6.3.7.2.Huntington's disease... .....r.r-1ii.i....,,.."..........".144
6,3.E Glial frbri'llary asidic piotein irnmunoreastivity in the
subependymal layer
6.3.8.1 Norma]..,........-..,......r.. .,....r.......q!r.,.,..............145
6,.3-8.2,[Iuntington's disease-.. .....-..........,145
6.3.9 Vimentin .ir[rnunoreaotivity, in the subependymal layer
6.3.9.1Nonbal'..........!.t...ir!..'i!.!...'.........
6.3-92 Huntington's disease... ......rii!.i........,,......,......147
6.3.10 Ferrifin irii[runoreaetivity in the subependymal layer
6.3.10;1 Nonrral..........i!.!.........r,..:................ ...............148
6.3.10"2 Huntingtoll's disease...... ...,r!.....r.i.....,......,.,..148
6.3.I I,GABAA u,l subunitirnmunoreaetivity in the subependymal layer
6.3.11.1 Normal"..... .,............,149
6.3. 1 1,2 Huntingbn's disease...,..
6.3..12 GABA.A p2, 3 subunit imrnunoreactivity in the subependynal layer
5.3.121 Normal...... r...i.....,..!,............,.....151
6,3.|2.2Huntington'sdisease.............'..'..i........'..|...'..;..|.
xlv

6.3.13 GABAAy2 subunit irinmunsreactivity in the sr,rbependymal laya
6.3.13,1 Nsrlnal,.....!.....'.,...i.!...1 ......,..-.....151
6"3.132 Huntington's disease... ,.,..i..i.....;............,...d,...i..."..,,."...."..152
6.4 Diseussion..............
6.4.1 The adult no'rraral and Huntington"s disease subependymal
layer contain sorne projection neurons ,.."..154
6.4.2T\e adtdt nornial and Huntington's disease subependymal
Iayor contain some interneurons................ .-..,..,..........156
64.3 The adult normal arrd fluntinglon's disease s.ubependymal
laye! contain some glial cells.. .........,....."..158
6:4.4The adult normal and Huntington's disease subependymal
layer contain GABA6 reoeptor subunits..,.-.-.,...?......,r.. ...............160

Chapter 2
LIST OF TABLES AND FIGURES
Table 2.1
Table of antibodies and the cell or cellular component detected by the antibody.............19
Figure 2.1
Stem and progenitor cells during development and in the adult brain. .............5
Figure 2.2
hogenitor cells during in vitro and in vivo expansion, and
during symmetric or asymmetric division .........,...........6
Figure 2.3
The subependymal layer and granule cell layer in the rodent brain.......... ........9
Figure 2.4
Diagrammatic representation of the mouse subependymal layer
(as described by Doetsch et al (1997))...... ...................21
Figure 2.5
Diagrammatic representation of the 3 types of lateral ventricle wall
cytoarchitecture (as described by Rodriguez-Perez. et al (2003)).... ................................24
Figure 2.6
Cell types in the striatum and the relationship of the striatuln to
the subependymal 1ayer.......... ....................36
Chanter 3
Table 3.1
Table of normal and Huntington's disease cases.......... .................44
Figure 3.1
Anatomy of the ependymal and subependymal region in the normal brain......................48
Figure 3.2
Cell types in the subependymal layer of the normal adult human brain............................50
Figure 3.3
Comparison of the thickness of the subependymal layer in normal and
Huntington's disease brains........ ...............54

Figure 3.4
Thickness of the subependymal layer in normal
and Huntington's disease brains........ .......55
Figure 3.5
Thickness of the subependymal layer in normal
and Huntington's disease (grade l-3) brains. ..............56
Figure 3.6
The subependymal layer in normal and Huntington's
disease (grades I and 3) brains........ ..........57
Figure 3.7
Number of cells/mm in the subependyrnal layer of normal
and Huntington's disease brains........ ........59
Figure 3.8
Number of cells/mm in the subependymal layer of normal and Huntington's
disease (grades l-3) brains. ......60
Figure 3.9
Cell types in the subependymal layer and ependymal layer of the
normal and Huntington's disease brain.......... ..............62
Figure 3.10
Comparison of the numbers of the different cell types present in the
normal and Huntington's disease (grades l-3) subependymal Iayer.................................63
Chapter 4
Table 4.1
Normal and huntington's disease cases used for Western blotting analysis......................79
Figure 4.1
The number of proliferating cells in the Huntington's disease subependymal
layer increase as the grade and CAG repeat length increase..... .....................83
Figure 4.2
Western blot analysis of normal and Huntington's disease
subependymal layer homogenates................. ..............87
Figure 4.3
Localisation of proliferating, neuronal and glial cells in the
Huntington's disease subependymal layer...... .............89
xvll

Figure 4.4
Neurogenesis and gliogenesis occur in the subependyrnal
layer in Huntington's disease... ...,.,..:......... ..................91
Figure 4.5
A. The number of fluorescently labelled pcNA positive cells in
the normal and Huntington's disease subependyirar rayer
B. The number of pcNA/plll-tubulin positive cells in the normal
and Huntington's disease subependyrnal layer...... ......g4
Figure 4.6
Neurogenesis in the normal brain subependymal 1ayer.......... .......95
Figure 4.7
Neurogenesis in the Huntington's disease subependymal layer...... ................96
Chapter 5
Table 5.1
Table of normal and Huntington's clisease cases examined for the
distribution of PCNA positive ceils in the subependymal |ayer.......... ..........1 l0
Table 5.2
ANOVA table for fixed effects................ ..................117
Figure 5.1
Diagram of a coronal section of the hurnan brain demonstrating
'Areas' within the subependymal |ayer.......... ..........:....... ...........114
Figure 5.2
Diagrrun of a lateral section of the human brain clemonstrating .Areas,,
'Regions' and 'Locations' within the subependyrnal 1ayer.......... .................1l5
Figure 5.3
Comparison of the number of pCNA positive cells in the
normal and Huntington's disease subepenclymal layer..-... ..........1l9
Figure 5.4
The number of PCNA positive celrs in each area (clorsal, middle, ventral)
of the subependymal layer in normal and Huntingron.s disease brains...........................120
Figure 5.5
The number of PCNA positive cells in each region (rostral, central caudal)
of the subependymal layer in normal and Huntington's disease brains........ ...................122
xvill

Figure 5.6
The number of PCNA positive cells in different locations of the
subependymal layer in normal and Huntington's disease brains........ ...........123
Chanter 6
Table 6.1
Normal and Huntington's disease cases examined for the presence
of mature cell markers in the subependymal layer...... ................L34
Table 6.2
Table summarising the staining demonstrated in the ependyma, subependyma
and caudate nucleus in normal and Huntington's disease brains........ ...........153
Figure 6.1
hojection neuron markers in the subependyrnal layer
of normal and Huntinston's disease brains........ ........138
Figure 6.2
Interneuron markers in the subependymal layer of normal
and Huntington's disease brains........ ......141
Figure 6.3
Glial markers in the subependymal layer of normal
and Huntinglon's disease brains........ ......146
Figure 6.4
GABAA receptor subunits in the subependymal layer of
normal and Huntington's disease brains........ ............150
Chapter 7
Figure 7.1
Possible method of targeting the subependymal layer for the delivery substances
that facilitate cell proliferation and differentiation.... ..................175
Figure 7.2
Diagram of the anatomy of the subependymal region in
the Huntington's disease brain showing the possible path of
migration for replacement cells moving fi'om the subependymal
layer to the caudate nucleus...... ...............I77
xix

APS
AraiC
bctr-z
BDhIF
BLBP
BrdU
Calr
ChAT
,{
CREB
CT
DAB
DNA
EGF
ENK
EPI
FGF.2
GABA.
GAD
GFAP
HD
IP,
IGF.1
u
LV
MEAO
Iv{nn
NCAM
NeuN
an'rmonium porsulfate
cytosi ne-p-D-arabj nofu rancsi de
B-ccll lymphoma protein-Z
brain derived neurofrophic factor
brain lipid binding protein
2-5 bromodeoxyiiridine
sahetinin
choline-acetyl transferase
caudate nuclzus
cyclio adenosine monophosphate response element binding protein
computed tomogr4phy
3, 3 diamino benzidine
deoxy-ribose nueleic acid
epidermal growth f,actor
onkephalin
egendymal layer
fi broblast growth factor-2
y-arrino butyric acid
glutfiric aeid decarboxyl ase
gl-iat fibrillay acidic protein
Huntington's disease
in ,apefltoneal
insulin-like growth facton- I
labelling index
lateratr venfiicle
middlc cerebral artery occlusion
rnagnetic resonance imaging
nelral cell adhesion molecule
neuronal nuclei
xx

NGF nerve griwth factor
NMDA N-methyl D-aspartate
NPY neuropepridq y
NSE nouron*pecific enolase
PErv parv:albumin
PBS phosphate brdfered saline
PCNA proliferatingcell nuolear andgen
PS.A,-NCAM polysiarylated-neurdoefladhesionmorecule
PVDF polyvinylidinerdi-fluoride
r,pm revolutions per minute
SEt eubeBendyrnal layer
SP substancep
STR s,g'iatr'ri
t time.
T6 total cell,eycietime
Ts ttrre sp-ent in the S-phase sf the cell cyc-le
TUI{EL transf,erase merrliated t4deoxyuridine triphosphate nick end labelling
UV ultra violet
VIM vfunentin
xxr