Jin-Young Jeong. (2024). Characterization of cerebrovascular pathology according to the origin of α-synuclein in the Parkinson’s disease mouse model. doi: 10.22677/THESIS.200000802155
Type
Thesis
Description
α-synuclein, Parkinson’s disease, Cerebrovasculature, The secretable mouse model
Table Of Contents
List of Contents Abstract ··································································································i List of contents ························································································ ii List of tables ··························································································· v List of figures ························································································· vi Ⅰ. Introduction 1.1 Parkinson’s disease ······································································· 1 1.1.1 Neuronal defects of Parkinson’s disease ······································ 1 1.1.2 Vascular defects of Parkinson’s disease ······································· 1 1.2 General characteristics of α-synuclein ··············································· 2 1.2.1 The structure of α-synuclein ······················································· 2 1.2.2 The physiological function of α-synuclein ······································ 2 1.2.3 The pathological contribution of α-synuclein ·································· 3 1.3 Need for a new PD mouse model ····················································· 3 1.3.1 α-synuclein expression in physiological and pathological conditions in the periphery ··············································································· 3 1.3.2 Secretion of α-synucleins to extracellular space ····························· 4 1.4 Main goals and key results ······························································ 4 Ⅱ. Materials and Methods 2.1 Mouse models ·············································································· 6 2.2 Blood vessel labeling and tissue clearing ··········································· 6 2.3 Immunohistochemistry (IHC) ··························································· 7 2.4 Protein sample preparation ····························································· 8 2.5 Western blotting············································································ 9 2.6 Dot blot for protein aggregation assay ··············································· 9 2.7 Cell proliferation analysis ······························································ 10 2.8 Imaging and analyses ·································································· 11 2.9 Capillary isolation and RNA purification ············································ 11 2.10 cDNA synthesis and quantitative real-time PCR (RT-qPCR) ··············· 12 2.11 BBB permeability test ································································· 12 2.12 Electrophysiology ······································································ 13 2.13 Plasmid and cloning ··································································· 13 2.14 Cell culture and transfection ························································ 14 2.15 Conditioned media preparation ····················································· 14 2.16 Behavior test ············································································ 14 2.16.1 Open field test ····································································· 14 2.16.2 Wire hanging test ································································· 15 2.16.3 Grip strength assay ······························································ 15 2.17 Statistical analysis ····································································· 15 Ⅲ Results Part1. Neuro-to-vascular view of α-synuclein pathology 3.1 Underdevelopment of the cerebrovascular network in the familial PD mouse model ······················································································ 16 3.2 In young PD mice, the pathological form of human α-synuclein is selectively expressed in neurons ·································································· 17 3.3 Overexpressed human α‑synuclein does not induce abnormal inflammatory responses in young PD mice ························································ 18 3.4 Neonatal PD mice exhibit reduced angiogenic potential ······················· 18 3.5 Overexpressed human α-synuclein reduces neuronal activity in PD neonates······························································································ 19 3.6 Activity-dependent angiogenic gene expression in the endothelial cells is altered in young PD mice ····························································· 20 Part 2. Peripheral-to-neuronal view of α-synuclein pathology 3.7 Enhanced extracellular secretion of human α-synuclein by adding a signal peptide ···················································································· 22 3.8 Generation of a mouse model in which α-synuclein levels are selectively increased in the blood ································································· 22 3.9 Analysis of pathological changes resulting from elevated α‑synuclein levels in blood ····················································································· 23 Ⅳ Discussion 4.1 Vascular alteration precedes neuronal defects in neurodegenerative diseases ······························································································ 24 4.2 Vascular changes in animal models of PD provide implications for disease progression··············································································· 25 4.3 Neuronal activity is a regulator of vascular formation ··························· 26 4.4 Neurovascular interactions in PD ···················································· 27 4.5 Aberrant expression of α-synucleins impairs neuronal activity ··············· 28 4.6 Ptprb as a possible mediator for reduced endothelial proliferation ·········· 28 4.7 A valuable mouse model for studying the role of extracellular α-synuclein 29 4.8 Origin of α-synuclein and its toxicity in the blood ································ 29 4.9 Passage of α-synuclein through the BBB ········································· 30 Ⅴ. Reference ························································································ 55
