목차

I. Introduction 1
A. Roles of non-neural cells in the intact brain 1
1. Astrocyte 1
2. Microglia 2
B. Repair of the injured brain of wild type mice 3
1. Repair processes of injured brain 3
2. Diverse factors that mediate repair of the injured brain 4
2.1 Intrinsic factors 4
2.2 Extrinsic factors 6
3. Cells that are involved in the injured brain 7
3.1 Neuron 7
3.2 Astrocyte 8
3.3 Microglia 9
3.4 Monocyte 10
4. Extracellular vesicles 12
C. Neurodegenerative disease and repair of brain injury 15
1. Parkinson's disease and Parkinson's disease-related genes 15
2. Parkinson's disease and brain repair 16
3. LRRK2 G2019S 18
4. LRRK2 G2019S and brain repair 19
D. Aims of the study 23
II. Materials and Methods 24
A. Animals 24
B. Stereotaxic injection 24
C. Magnetic resonance imaging (MRI) 25
D. Skin wound injury 26
E. Tissue preparation 26
F. Immunostaining and image analysis 27
G. Preparation and analysis of exosome fractions 32
H. Electron microscopy 34
I. Fluorescence-activated cell sorting (FACS) 34
J. Cultures of neurons, astrocytes, microglia and macrophages 35
K. Neurite outgrowth assay 37
L. Organotypic cortical slice culture 37
M. LDH assay 37
N. Western blot analysis 38
O. Quantitative real-time PCR 39
P. Transfection and drug treatment 40
Q. Statistical analysis 40
III. Results 42
A. Brain injury and repair in wild type mice 42
1. Damaged neurons in the injured brain express OPN and become fragmented into extracellular particles 42
2. Infiltrated monocytes engulf and fragment neuronal cell bodies 53
3. Fragmented neuronal cell bodies guide the growth of neurites and astrocyte processes toward the injury core 58
4. Exosomes prepared from the injured brain enhance neurite outgrowth 65
B. Brain injury and repair in LRRK2 G2019S mice 70
1. LRRK2 G2019S attenuates the repair processes after injury 70
2. LRRK2 G2019S reduces the number of extracellular vesicles and their OPN levels 76
3. LRRK2 G2019S alters monocyte pyroptosis but not infiltration after brain injury 88
4. Insufficient OPN-positive particles hamper elongation of neurites and astrocyte processes in LRRK2 G2019S mice 99
5. Exosomes containing OPN restore neurite outgrowth in LRRK2 G2019S neurons 103
IV. Discussion 107
A. Brain injury and repair in wild type mice 107
B. Brain injury and repair in LRRK2 G2019S mice 116
V. Conclusion 123
Reference 125