Microglial migration and adhesion molecules during embryonic brain development

218 Pages·2017·8.78 MB·English

by  Sophie Smolders

Microglial migration and adhesion molecules during
embryonic brain development
Sophie Smolders
To cite this version:
Sophie Smolders. Microglial migration and adhesion molecules during embryonic brain development.
Neurons and Cognition [q-bio.NC]. Université Pierre et Marie Curie - Paris VI; Universiteit Hasselt
(1970-..), 2017. English. ￿NNT: 2017PA066533￿. ￿tel-01890177￿
HAL Id: tel-01890177
https://theses.hal.science/tel-01890177
Submitted on 8 Oct 2018
HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est
archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents
entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non,
lished or not. The documents may come from émanant des établissements d’enseignement et de
teaching and research institutions in France or recherche français ou étrangers, des laboratoires
abroad, or from public or private research centers. publics ou privés.
Université Pierre et Marie Curie
Universiteit Hasselt
CERVEAU - COGNITION - COMPORTEMENT
Neurosciences Paris Seine, UPMC INSERM S 1130, CNRS UMR 8246
Development of Spinal Cord Organization

La migration des microglies et les molécules adhésives au
cours du développement embryonnaire du cerveau

Par Sophie Smolders
Thèse de doctorat de Neurosciences
Dirigée par Pascal Legendre et Bert Brône
Présentée et soutenue publiquement le 30 octobre 2017
Devant un jury composé de :
Prof. dr Marcel Ameloot, Universiteit Hasselt, Diepenbeek, Belgium, Maître de conférences
Prof. dr. Bert Brône, Universiteit Hasselt, Diepenbeek, Belgium, promotor
Dr. Pascal Legendre, Université Pierre et Marie Curie, Paris, France, promotor
Prof. dr. Niels Hellings, Universiteit Hasselt, Diepenbeek, Belgium
Dr. Isabelle Dusart, Université Pierre et Marie Curie, Paris, France
Dr. Michel Mallat, Université Pierre et Marie Curie, Paris, France
Dr. Etienne Audinat, Université Paris Descartes, Paris, France, rappoteur
Prof. Dr. Charles ffrench-Constant, University of Edinburgh, Edinburgh, UK, rapporteur

“What we observe is not nature itself,
but nature exposed to our method of questioning”

Werner Heisenberg

Microglia. Río-Hortega, 1919 & 1921

Table of contents
Table of contents I
List of Abbreviations V
CHAPTER 1: 1
General introduction and aims
1.1 Basics of embryonic brain development 3
1.1.1 Progenitor amplification: E9-E12 3
1.1.2 Neurogenesis and cortical layering: E11-E18 4
1.1.3 Angiogenesis: from E8.0 4
1.1.4 Gliogenesis: E16-P20 5
1.2 The microglial cell: versatile outsider and one of a kind 6
1.2.1 Origin of microglia 7
1.2.2 Migration into the CNS 9
1.2.3 Microglial maturation inside the CNS 15
1.2.4 Homeostasis of microglia in the adult CNS 16
1.2.5 Undersigned, I, the microglial cell 17
1.2.6 Parenchymal and non-parenchymal CNS macrophages 18
1.2.7 Physiological functions of microglia 22
1.2.8 Functions of microglia in pathology 26
1.2.9 Gender differences 30
1.3 Cell Migration 31
1.3.1 ECM 32
1.3.2 Integrins 35
1.3.3 ECM/integrin signaling in microglia and their mobility 39
1.4 Study aims 42
CHAPTER 2: 45
Complex invasion pattern of the cerebral cortex by microglial cells during development of the mouse
embryo
2.1 Abstract 46
2.2 Introduction 46
2.3 Materials and methods 47
2.4 Results 50
2.5 Discussion 62

I
CHAPTER 3: 65
Age-specific function of α5β1 integrin in microglial migration during early colonization of the
developing mouse cortex
3.1 Abstract 66
3.2 Introduction 66
3.3 Materials and Methods 67
3.4 Results 72
3.5 Discussion 87
CHAPTER 4: 95
The CXCL12/CXCR4/integrinß1 signaling axis in microglial migration: an exploratory study
4.1 Abstract 96
4.2 Introduction 96
4.3 Materials and Methods 98
4.4 Results 102
4.5 Discussion 114
4.6 Conclusion 119
4.7 Supplemental data description 119
CHAPTER 5: 129
Maternal immune activation evoked by polyinosinic:polycytidylic acid does not evoke microglial cell
activation in the embryo
5.1 Abstract 130
5.2 Introduction 130
5.3 Materials and Methods 132
5.4 Results 134
5.5 Discussion 145
CHAPTER 6: 149
General discussion and perspectives
6.1 Properties of microglia during colonization of the embryonic brain 150
6.1.1 Morphology and protein markers 151
6.1.2 Maturation and the microenvironment 152
6.2 The microglial migration behaviour in the embryonic brain 152
6.2.1 Microglial settlement during embryogenesis 153
6.2.2 Regulation of microglial migration speed 154
6.3 Complex regulation of integrin function in microglial migration during development 155
6.4 Candidate cues for steering parenchymal microglial migration during development 156
II
6.4.1 Chemoattractants and chemorepellants 156
6.4.2 Cell death and synaptogenesis 157
6.5 Microglial interactions with blood vessels 158
6.5.1 Mechanisms of interaction 158
6.5.2 Attraction towards blood vessels 158
6.5.3 Mechanisms of influencing blood vessel branching 159
6.6 Does microglial activation by maternal immune activation lead to neurodevelopmental disorders? 160
6.6.1 Priming of microglia 160
6.6.2 Reported alterations in microglia upon MIA 161
6.6.3 Controversies in microglial activation following MIA 161
6.7 Limitations of this work 162
6.8 Outstanding questions 164
List of references 165
Résumé 189
Nederlandse samenvatting 191
English summary 193
Curriculum Vitae 195
Dankwoord - Acknowledgments 199
III

List of Abbreviations
Akt Serine/threonine kinase
ASD Autism spectrum disorder
ATP Adenosin triphosphate
BBB Blood-brain barrier
BDNF Brain derived neurotrophic factor
BP Bandpass
BSA Bovine serum albumin
C Complement
CCL C-C motif chemokine
CCR C-C motif chemokine receptor
CD Cluster of differentiation
CNS Central nervous system
CNS Central nervous system
CNTF Cilliary neurotrophic factor
CR Complement receptor
CSF (R) Colony stimulating factor (receptor)
CSPGs Chondroitin sulphate proteoglycans
CT-1 Cardiotrophin-1
CX3CR1 Chemokine (C-X3-C motif) receptor 1
CXCL C-X-C motif chemokine
CXCR C-X-C motif chemokine receptor
DAP12 DNAX activation protein of 12kDa
DAPI 4',6-diamidino-2-phenylindole
DMEM Dulbecco's Modified Eagle's Medium
E Embryonic day
ECM Extracellular matrix
eGFP Enhanced green fluorescent protein
ERK Extracellular signal–regulated kinases
FAK Focal adhesion kinase
FBS Fetal bovine serum
FN Fibronectin
GABA γ-aminobutyric acid
GDNF Glial derived neurtrophic factor
GE Ganglionic eminences
GFAP Glial fibrillary acidic protein
GLAST Glutamate aspartate transporter
(G)M-CSF(R) (Granulocyt) macrophage coloniy-stimulating factor
GTP Guanine triphosphate
HBSS Hank's Balanced Salt Solution
HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
HIV-1 Human type 1 immunodeficiency virus
HS Horse serum
HSC Hematopoietic stem cell
Iba-1 Ionized calcium-binding adapter molecule 1
ICAM-1 Intercellular Adhesion Molecule 1
IFN Interferon
IGF Insulin-like growth factor
IL Interleukin
iNOS Inducible nitric oxide synthase
iPLA2 Calcium-independent phospholipase A2
KO Knockout
LFA Lymphocyte function-associated antigen
LGE Lateral ganglionic eminence
V