Regulation of adult neurogenesis in Huntington s disease [Elektronische Ressource] : the role of TGF-beta1 signaling in the neurogenic niche / Mahesh Kandasamy
129 pages
English

Regulation of adult neurogenesis in Huntington's disease [Elektronische Ressource] : the role of TGF-beta1 signaling in the neurogenic niche / Mahesh Kandasamy

-

Le téléchargement nécessite un accès à la bibliothèque YouScribe
Tout savoir sur nos offres
129 pages
English
Le téléchargement nécessite un accès à la bibliothèque YouScribe
Tout savoir sur nos offres

Description

Regulation of adult neurogenesis in Huntington’s disease: The role of TGF-beta1 signaling in the neurogenic niche DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES DER NATURWISSENSCHAFTEN (DR. RER. NAT.) DER FAKULTÄT FÜR BIOLOGIE UND VORKLINISCHE MEDIZIN DER UNIVERSITÄT REGENSBURG vorgelegt von Mahesh Kandasamy aus Dharmapuri, India 2010 Das Promotionsgesuch wurde eingereicht am: 24.09.2010 Die Arbeit wurde angeleitet von: Prof. Dr. Inga Neumann und Prof. Dr. Ludwig Aigner in der Klinik und Poliklinik für Neurologie der Universität Regensburg Table of Contents 1. Introduction .......................................................................................................................... 1 1.1. Adult neurogenesis ................. 1 1.2. History of adult neurogenesis ................................ 2 1.3. Stem cells in the adult brain ................................................................... 3 1.4. Hippocampal neurogenesis ..... 4 1.5. Neurogenesis in the SVZ-RMS-OB system .............................................. 5 1.6. Regulation of adult neurogenesis............................................................ 7 1.7. Regulation of neurogenesis by signaling molecules .............................................................. 8 1.8. Transforming growth factors ...................................

Sujets

Informations

Publié par
Publié le 01 janvier 2011
Nombre de lectures 25
Langue English
Poids de l'ouvrage 3 Mo

Extrait


Regulation of adult neurogenesis in Huntington’s disease:
The role of TGF-beta1 signaling in the neurogenic niche




DISSERTATION ZUR ERLANGUNG DES DOKTORGRADES
DER NATURWISSENSCHAFTEN (DR. RER. NAT.)
DER FAKULTÄT FÜR BIOLOGIE UND VORKLINISCHE MEDIZIN
DER UNIVERSITÄT REGENSBURG



vorgelegt von
Mahesh Kandasamy
aus
Dharmapuri, India






2010

Das Promotionsgesuch wurde eingereicht am: 24.09.2010
Die Arbeit wurde angeleitet von: Prof. Dr. Inga Neumann und Prof. Dr. Ludwig Aigner
in der Klinik und Poliklinik für Neurologie der Universität Regensburg

































Table of Contents

1. Introduction .......................................................................................................................... 1
1.1. Adult neurogenesis ................. 1
1.2. History of adult neurogenesis ................................ 2
1.3. Stem cells in the adult brain ................................................................... 3
1.4. Hippocampal neurogenesis ..... 4
1.5. Neurogenesis in the SVZ-RMS-OB system .............................................. 5
1.6. Regulation of adult neurogenesis............................................................ 7
1.7. Regulation of neurogenesis by signaling molecules .............................................................. 8
1.8. Transforming growth factors ................................... 9
1.8.1. The TGF-beta signaling pathway ...................................................... 10
1.8.2. TGF-beta expression in the normal and pathological brain .............................................. 11
1.8.3. TGF-beta signal transduction in the brain ........................................ 12
1.8.4. Roles of TGF-beta in the brain.......................................................... 13
1.8.5. Regulation of adult neurogenesis by TGF-beta1 .............................. 13
1.8.6. Enhanced TGF-beta 1 level and impaired neurogenesis in neurodegenerative disorders 14
1.9. Huntington’s disease............................................................................................................. 15
1.9.1. Clinical aspects of Huntington’ s disease ......... 15
1.9.2. History of Huntington’s disease ....................... 16
1.9.3. Epidemiology of Huntington’s disease ............................................................................. 17
1.9.4. Localization and function of the physiological Huntingtin protein ... 18
1.9.5. Expanded CAG repeats in the huntingtin gene................................................................. 19
1.9.6. Neuropathological hallmarks of HD .................................................................................. 20
1.9.7. Experimental models of Huntington’s disease . 21
1.9.7.1. Acute models for Huntington’s disease......... 21
1.9.7.2. Transgenic models of Huntington’s disease ................................................................. 21
1.9.8. Neurogenesis in Huntington’s disease ............ 22

2. Aim of the study ................................................................................................................ 23

3. Material and Methods....... 24

3.1. Materials .............................................................................................................................. 24
3.1.1. Expendable materials ....................................... 24
3.1.2. Chemicals for in vivo immunological procedure 25
3.1.3. Chemicals for Western blot .............................................................................................. 26
3.1.4. Cell culture medias ........................................... 26
3.1.5. Other reagents for cell culture.......................................................................................... 27
3.1.6. Buffer, solutions and stock solutions............... 28
3.1.7. Primary antibodies ........................................................................................................... 31
3.1.8. Secondary antibodies ...... 32
3.1.7. Devices............................................................................................................................. 32
3.1.8. Software ........................... 33

3.2. Methods............................................................................................................................... 34
3.2.1. Animals....... ...................... 34
3.2.2. Intracerebroventricular infusions of TGF-beta1................................ 34
3.2.3. BrdU labelling................................................................................................................... 35
3.2.4. Tissue processing and immunohistochemistry 35
3.2.5. Counting procedures........ 37
3.2.6. Western Blotting............................................................................................................... 39
3.2.7. Neural stem and progenitor cells culture ......... 40
3.2.8. Cell cycle analysis ............ 41
3.2.9. Immunocytochemistry ...................................................................................................... 41
3.3. Statistics ............................................................. 42

4. Results ................................................................................................. 44
4.1. TGF-beta1 signaling components are expressed throughout the adult rat brain.................. 44
4.2. Table 1-Semiquantitative measurement of immunoreactivity of TGF-betaRII, TGF-betaRI and
pSmad2 in the adult rat brain ............................................................................................. 49
4.3. pSmad2 is predominantly present in postmitotic cells in the hippocampus of the adult
brain. .................................................................... 51
4.4. Induced over-expression of TGF-beta1 in the hippocampus reduces cell proliferation but
promotes neuronal differentiation and survival.. .................................. 54
4.5. Elevated levels of TGF-beta1 provokes expression of pSmad2 in neural stem and
progenitor cells .................................................................................................................... 57
4.6. Regulation of hippocampal neurogenesis in tgHD rats ........................ 61
4.7. Hippocampal cell proliferation in tgHD rats gets progressively impaired between 8 and 12
months of age....................................................................................................................... 61
4.8. Impaired survival of newly generated cells and reduced neuronal density was mediated by
reduced CREB signaling in tgHD rat hippocampus. ............................. 61
4.9. Increased quiescence of newly generated cells in tgHD dentate gyrus ................................ 65
4.10. Neuroblast proliferation compensate s stem cell quiescence in tgHD dentate gyrus........... 67
4.11. Reduced cell proliferation in tgHD animals correlates with increased TGF-beta1 signaling
in hippocampal stem cells.................................................................................................... 70



5. Discussion .......................................................................................................................... 77

7. Bibliography ....................... 88

Acknowledgements ............................................................................................................ 118



List of abbreviations
-MEM Alpha Modified Eagle Media
AD Alzheimer's disease
ALK Activin like kinase
BDNF Brain Derived Neurotrophic Factor
Bmi-1 B lymphoma Mo-MLV insertion region-1
BMP-2 Bone Morphogenetic Protein 2
BMP-4 Bone Morphogenetic Protein 4
bp Base pair
BSA Bovine Serum Albumine
BrdU 5-bromo-2-deoxyuridine
BT Biotinylated
CAG Cytosine, Adenine and Guanine
CBP CREB-binding protein
CNTF Ciliary neurotrophic factor
Cor Cortex
CNS Central Nervous System
CREB cAMP response element-binding
CSF Cerebrospinal fluid
ACSF Artificial Cerebrospinal fluid
DAPI 4’,6-Diamidino-2-phenylindole
DG Dentate Gyrus
DCX Doublecortin
DNA Deoxyribonucleic acid
DNI Dystrophic Neuritic Inclusions
DMEM Dulbecco’s Modified Eagle Media
EDTA Ethylenediaminetetraacetic acid
EGF Epidermal Growth Factor
ES cell Embryonic Stem cell
EPO Erythropoietin
FBS Fetal Bovine Serum
FGF Fibroblast Growth Factor
FSGB Fish Skin Gelatin Buffer
GABA Gama Aminobutyric acid
GCL Granule Cells Layer
G-CSF Granulocyte colony-stimulating factor
GDF Growth differentiation factors
GFAP Glial Fibrillary Acidic Protein
GS-Domain Glycin-Serine domain
IGF-1 Insulin-like Growth Factor 1
h Hour
HC Hippocampus
HD Huntington’s disease
Hes Hairy and enhancer of spl

  • Univers Univers
  • Ebooks Ebooks
  • Livres audio Livres audio
  • Presse Presse
  • Podcasts Podcasts
  • BD BD
  • Documents Documents