Hyeji Jung. (2025). Investigation of Regional and Temporal Dynamics of Microglial Activation: Impacts of Dysregulated NLRP3 Inflammasome on Brain Function. doi: 10.22677/THESIS.200000841428
Type
Thesis
Description
신경염증, 미세아교세포, 사이토카인, NLRP3, 시냅스, NMDA 수용체
Table Of Contents
Ⅰ. INTRODUCTION 1 1.1 General introduction: Diversity building up the brain 1 1.2 Core structure of excitatory and inhibitory synapses and their diversity 3 1.3 Contribution of microglia to sophisticated brain function 4 1.4 Neuroinflammation and its impact on microglial function 5 1.5 Regulatory mechanisms of the NLRP3 inflammasome and the pathological contribution of its dysregulation 5 II. MATERIALS & METHODS 7 2.1 Animals 7 2.2 Drug treatment 7 2.3 Antibodies 7 2.4 Immunohistochemistry (IHC) 8 2.5 Microglial analyses using slide scanner 9 2.6 Microglial morphology analyses 9 2.7 Microglial 3D analyses 9 2.8 Preparation and titration of AAVs 10 2.9 Stereotaxic injections 10 2.10 Analysis of microglial engulfment using a synapse engulfment reporter virus 10 2.11 Preparation of brain lysates and quantification of cytokine expression levels 11 2.12 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) 11 2.13 Primary microglia culture 12 2.14 Culture, immunostaining, and image analysis of cultured neurons 13 2.15 Behavior tests 13 2.16 Data analysis and statistics 15 III. RESULTS 16 3.1 Brain region-specific microglial response to LPS-induced neuroinflammation 16 3.2 Temporal dynamics of microglial activity and its impact on GABAergic synapses in LPS-induced neuroinflammation 19 3.3 NLRP3 dysregulation in microglia induced exaggerated and prolonged neuroinflammation to disrupt normal brain functions. 21 IV. DISCUSSION & CONCLUSION 28 4.1. Regional discrepancy in microglial activation during neuroinflammation 28 4.1.1 Susceptibility of Hb to mild neuroinflammation 28 4.1.2 Different clusters of brain regions show relatively severe responses to two daily doses of LPS 29 4.1.3 Different clusters of brain regions show relatively higher CD68 expression in response to a single dose of LPS. 30 4.1.4 Regional discrepancy in cytokine expression 30 4.2. Investigation of temporal dynamics of microglial activation 32 4.2.1 Microglial activation resolves across time. 32 4.2.2 Inhibitory synapse-specific impairment upon systemic LPS administration ·· 33 4.3 Effects of NLRP3-Dependent neuroinflammation on brain function 34 4.3.1 Abnormal microglial phenotypes induced by NLRP3 dysregulation lead to prolonged and intensive neuroinflammation. 35 4.3.2 Excitatory postsynaptic structures and properties get altered in severe neuroinflammation but not in mild neuroinflammation. 36 4.3.3 Excessive NMDAR response in neuroinflammatory brain dictates abnormal repetitive behavior of mice. 37 V. REFERENCE 87
