Properties of nestin-GFP-expressing cells in different regions of adult murine brain [Elektronische Ressource] / von Liping Wang

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• Publié par: charite_-_universitatsmedizin_berlin
• Publié le: 01 janvier 2005
• Nombre de lectures: 34
• Langue: English
• Poids de l'ouvrage: 7 Mo

Extrait

Aus dem International Graduate Program Medical Neurosciences
der Medizinischen Fakultät der Charité – Universitätsmedizin Berlin




DISSERTATION


Properties of Nestin-GFP-Expressing Cells in
Different Regions of Adult Murine Brain




Zur Erlangung des akademischen Grades
Philosophiae Doctor (PhD)




vorgelegt der Medizinischen Fakultät der Charité –
Universitätsmedizin Berlin





von


Liping Wang

Hebei,China Properties of Nestin-GFP-Expressing Cells in Adult Brain 2
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Dekan: Prof. Dr. med. Martin Paul


Betreuer:


Gutachter: 1. Prof.Dr.Helmut Kettenmann
2. Prof.Dr.Christian W. Steinhäuser
3. Prof.Dr.Joachim W. Deitmer




Datum der Promotion: 18.July.2005




2 Properties of Nestin-GFP-Expressing Cells in Adult Brain 3
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Acknowledgements

I would like to thanks my supervisor Prof. Dr. Helmut Kettenmann for giving me the
opportunity to work on this particular subject in his laboratory. I am grateful for all the
support and advice.

I also wish to thank PD Dr. Gerd Kempermann, PD Dr. Matthias Endres and Prof. Dr.
Herbert Zimmermann for the fruitful collaboration. Especially I want to thank Dr. Golo
Kronenberg and Dr.Norbert Braun for offering me some unpublished data supporting the
discussions in the thesis.

Supporting in experiments, I would like to thank Dr. Vitali Filippov, Dr. Rene Jüttner and
Carola Schipke for their helpful suggestions.

Furthermore…
If I would like to name everybody who has kindly contributed to my PhD study and my
thesis, the list would be endless. Thus, I apologize to everybody reading these lines not be
mentioned personally.

Special thanks go to the International Graduate Program Medical Neuroscience of the
Berlin Medical University, Charité, Germany. The Graduate Program "Dynamics and
Evolution of Cellular and Macromolecular Processes" of Humboldt University of Berlin,
Germany. This work was supported by grants of the German Research Association
(Graduiertenkolleg 268 to L.P.W. and SFB 515).











3 Properties of Nestin-GFP-Expressing Cells in Adult Brain 4
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TABLE OF CONTENTS

ACKNOWLEDGEMENTS 3
TABLE OF CONTENTS 4
INDEX OF FIGURES 8

ABBREVIATIONS 11

ABSTRACTS 12

1. INTRODUCTION
1.1. Endogenous Adult Neural Stem Cells 16
1.1.1. Adult neurogenesis in hippocampus 16
1.1.2. Adult Neurogenesis in SVZ 20
1.1.3. Adult neurogenesis in other areas of the adult brain 21
1.2. Regulation of adult neurogenesis 22
1.3. Expression of neurotransmitters receptors are involved in neurogenesis 25
1.3.1. γ -Aminobutyric acid (GABA) Receptors 26
1.3.1.1. GABA receptors expression in NSCs 26
1.3.1.2. GABA receptors expression in neuroblasts 27
1.3.1.3. GABA receptors expression in glioblasts 28
1.3.2. Glutamate Receptors 29
1.3.2.1. Glutamate receptors expression in NSCs 30
1.3.2.2. Glutamate receptors expression in NPCs 31
1.3.2.3. Glutamate receptors expression in glioblasts 33
1.4. Neurogenesis following ischemic brain insults 34
1.4.1. Ischemic injury promote NPCs proliferation 35
1.4.2. NPCs migrate from proliferative zones to damaged regions following
ischemic insult 36
1.4.3. Differentiation and maturation of NPCs after ischemic insult 37
1.4.4. Regulation of adult neurogenesis after ischemic insults 37
1.5. Reactive astrocytes following the brain injury
1.5.1. Bi-directional communication between neuron and glia 38
1.5.2. Reactive astrogliosis following the brain injury 40
4 Properties of Nestin-GFP-Expressing Cells in Adult Brain 5
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2. EXPERIMENTAL PROCEDURES
2.1. Animals 41
2.2. Preparation of acute brain slices 42
2.3. Identification of the nestin-GFP-expressing cells 42
2.4. Materials 44
2.5. Electrophysiological recordings
2.5.1. Establishing whole-cell recording configuration 44
2.5.2. Study of conductance change induced by neurotransmitter during whole
cell recording 46
2.5.3. Synaptic currents recording 47
2.5.4. Fluorescent dye dialyzed nestin-GFP-expressing cells during
whole cell recording 47

3. RESULTS
3.1. Two distinct subpopulations of nestin-expressing progenitor cells were
distinguishable in adult mouse dentate gyrus 48
3.2. Earliest synaptic input during adult hippocampal neurogenesis is GABAergic
3.2.1. The non-radial precursor cells can express neuronal properties 48
3.2.2. Stimulation of the perforant path triggers GABAergic synaptic currents in a small
population of the non-radial precursors 51
3.2.3. Both types of nestin-GFP-expressing cells respond to GABA 56
3.2.4. Nestin-GFP-expressing cells express functional glutamate receptors of the AMPA
subtype 56
3.2.5. Nestin-expressing precursor cells express functional glutamate transporters 58
3.3. Functional expression of the nucleotide receptors by nestin-expressing
precursor cells in the adult Murine hippocampus 60
3.4. Membrane current features and functional expression of AMPA/Kainate
receptors by nestin-GFP-expressing cells in the adult amygdala
3.4.1. Membrane current features of Nestin-GFP-expressing cells in amygdala 61
3.4.2. Nestin-GFP-expressing cells in amygdala express functional glutamate
receptors of the AMPA/Kainate subtype in situ 62
3.5. Membrane current features of nestin-GFP-expressing cells in alveus CA1 and
corpus callosum
3.5.1. Nestin-GFP-expressing also be found in corpus callosum, CA1 and alveus 68
5 Properties of Nestin-GFP-Expressing Cells in Adult Brain 6
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3.5.2. Astrocytic properties in nestin-GFP-expressing cells in alveus and CA1 69
3.5.3. Properties of small nestin-GFP-expressing bipolar cells in alveus and
corpus callosum 71
3.5.4. Nestin-GFP-expressing cells with large bipolar processes oriented
perpendicular to the corpus callosum/ pyramidal cell layer are characterized
by passive membrane currents 72
3.6. Membrane current features and functional expression of AMPA/Kainate
receptors by nestin-expressing precursor cells in the adult striatum
after stroke
3.6.1. Nestin-GFP-expressing cells acquire the ‘complex’ physiological phenotype
after stroke 74
3.6.2. The ´complex cells´ express AMPA/kainate glutamate receptors, but lack
glutamate transporter currents 77


4. DISCUSSION
4.1. One subpopulation of nestin-expressing progenitor cells share astrocytic
features in the adult Murine hippocampus 81
4.2. Earliest synaptic input during adult hippocampal neurogenesis
is GABAergic 82
4.2.1. The type-2 nestin-positive population contains the earliest form of neurons during
maturation 83
4.2.2. Adult-generated neurons first receive GABAergic input 84
4.3. Functional relevance of ecto-nucleotidase activity and nucleotide receptors
in adult neurogenesis 85
4.4. Nestin-GFP-expressing cells display OPC features in amygdala 90
4.5. -expressing cell display astrocytes features from corpus
calosum toCA1 92
4.6. Nestin-GFP-expressing cells acquire the ´complex´ physiological phenotype
after stroke and express AMPA/kainate glutamate receptors currents 95
4.6.1. Nestin-GFP-expressing cells were upregulated following 30 min
MCAo/ reperfusion, but Nestin-GFP-expressing cells and GFAP+ cells are
distinct cellular populations 96
4.6.2. Nestin-GFP-expressing cells proliferate in situ 100
6 Properties of Nestin-G