The mechanistic action of IQSEC3 deletion in hippocampal dentate gyrus on network dysfunction
- Title
- The mechanistic action of IQSEC3 deletion in hippocampal dentate gyrus on network dysfunction
- Alternative Title
- 억제성 시냅스 단백질 IQSEC3 와 뇌전증
- Author(s)
- Dongseok Park
- DGIST Authors
- Dongseok Park ; Ji Won Um ; Eunji Cheong
- Advisor
- 엄지원
- Co-Advisor(s)
- Eunji Cheong
- Issued Date
- 2021
- Awarded Date
- 2021/02
- Type
- Thesis
- Table Of Contents
-
I. Introduction 1
1.1 Introduction 1
1.2 Inhibitory synapse organizers 4
1.3 Synaptic scaffolding protein IQSEC family 4
1.3.1 IQSEC1 6
1.3.2 IQSEC2 7
1.3.3 IQSEC3 8
1.4 IQSEC3 as a GEF for ARF1/6 10
1.5 Principle of neuron excitation 11
1.6 Neuronal circuitry of generalized seizure 12
1.7 Epilepsy and microglia 14
1.8 Non-inflammatory role of microglia 19
1.9 Potential role of IQSEC3 in epileptogenesis 20
II. MATERIALS AND METHODS 21
2.1 Materials 21
2.1.1 Chemicals and Drugs 21
2.1.2 Antibodies 21
2.1.3 Recombinant DNA 21
2.1.4 Software 22
2.2 Methods 22
2.2.1 Animal care 22
2.2.2 HEK293T Cell culture 22
2.2.3 Competent cell preparation 23
2.2.4 Recombinant plasmid purification 24
2.2.5 Adeno-associated virus preparation 24
2.2.6 Virus titer measure 25
2.2.7 Stereotactic virus injection 26
2.2.8 Brain perfusion and extraction 26
2.2.9 Sectioning of the brain 27
2.2.10 Immunohistochemistry 27
2.2.11 Confocal microscopy 29
2.2.12 Synapse puncta analysis 29
2.2.13 qRT-PCR 30
2.2.14 Head-mounted electrode preparation for EEG recording 31
2.2.15 Stereotactic surgery of head-mounted electrode 31
2.2.16 Electroencephalogram recording 33
2.2.17 Ictal seizure and interictal event analysis 33
2.2.18 Cell type specific SST peptide expression 34
2.2.19 Drug treatment 36
2.2.20 Morphology analysis of microglia 38
2.2.21 Microglial density analysis 38
2.2.22 Statistical analysis 38
III. RESULTS 39
3.1 IQSEC3 loss impairs structures and function of GABAergic synapses 39
3.1.1 IQSEC3 is necessary to maintain GABAergic synapses 39
3.1.2 IQSEC3 loss induces spontaneous recurrent seizure 48
3.2. Impaired GABAergic synapse induces epilepsy 54
3.2.1 Changes in epileptic brains 54
3.2.1.1 Reduced SST peptide expression 54
3.2.1.2 Activated microglia 57
3.2.2 Inhibition of microglial activation alleviates epilepsy progression 61
3.2.2.1 SST peptide expression reduces seizure susceptibility 61
3.2.2.2 Minocycline treatment reduces seizure severity 63
IV. DISCUSSION & CONCLUSION 66
- URI
-
http://dgist.dcollection.net/common/orgView/200000364865
http://hdl.handle.net/20.500.11750/16706
- Degree
- Doctor
- Department
- Brain and Cognitive Sciences
- Publisher
- DGIST
- Related Researcher
-
-
Um, Ji Won
- Research Interests Molecular and cellular mechanisms underlying synapse elimination; Key synaptic mechanisms associated with Alzheimer's disease and autism spectrum disorders; Synaptic homeostasis
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- Appears in Collections:
- Department of Brain Sciences Theses Ph.D.
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