Title

Modulation of Kv7.2/7.3 Channels and Membrane Excitability in Neuronal Cells

Alternative Title

전압 개폐성 7.2/7.3 포타슘 이온통로의 조절과 신경세포내의 막흥분성

DGIST Authors

Kwon-Woo Kim ; Byung-Chang Suh ; Hyun-Ho Lim

Advisor

서병창

Co-Advisor(s)

Hyun-Ho Lim

Issued Date

2021

Awarded Date

2021/02

Citation

Kwon-Woo Kim. (2021). Modulation of Kv7.2/7.3 Channels and Membrane Excitability in Neuronal Cells. doi: 10.22677/thesis.200000364452

Type

Thesis

Subject

Kv7.2/7.3 channel, ethanol, microRNA, SCG neuron, Hippocampal neuron, PI(4,5)P2, 전압 개폐성 7.2/7.3 포타슘 이온통로, 에탄올, 마이크로RNA, 상위의 자궁경부 신경세포, 해마 신경세포, PI(4,5)P2

Table Of Contents

I. Introduction 1
II. MATERIALS AND METHODS 9
Cell culture and transfection 9
Dissociation of superior cervical ganglia 9
Electrophysiological recording 10
Confocal imaging and analysis 11
Förster Resonance Energy Transfer (FRET) 11
Solution and materials 12
Non-stationary noise analysis 12
Animals and tissue collection 13
Dissection and dissociation of primary hippocampal neurons 14
Western blot analysis 14
RNA extraction and real-time PCR assays for miRNA and mRNA 15
Plasmid construction 16
Image acquisition 16
Data analysis and statistical analysis 17
III. RESULTS 18
3.1. Ethanol Elevates Excitability of Superior Cervical Ganglion Neurons by Inhibiting Kv7 Channels in a Cell Type-Specific and PI(4,5)P2-Dependent Manner 18
SCG neurons can be classified into phasic-1, phasic-2, and tonic neurons based on their action potential firing patterns and IKv7 size 18
Excitability and the resting membrane potential of SCG neurons are affected by inhibiting Kv7 channels in a cell type-specific manner 21
Ethanol increases the excitability of SCG neurons 23
Ethanol inhibits Kv7 channels in cultured SCG neurons as well as the heterologous Kv7.2/7.3 channels expressed in tsA201 cells 26
The inhibition of IKv7 by ethanol is not due to either degradation or translocation of plasma membrane PI(4,5)P2 29
Inhibitory effect of ethanol on IKv7.2/7.3 is modulated by the level of the plasma membrane PI(4,5)P2 31
Ethanol inhibition of the Kv7.2/7.3 current is the result of reduction in PI(4,5)P2 binding affinity of the Kv7.2/7.3 channel 36
Ethanol inhibition of Kv7.2/7.3 channels differs from block with internal TEA 40
Ethanol reduces the open probability of the Kv7.2/7.3 channel 44
3.2. Post-transcriptional control of KCNQ2 gene expression in brain by miRNAs 47
Kv7.2 channel expression is up-regulated by miR-106b family during early developmental stage in mouse hippocampus 47
M-currents are inhibited by up-expression of miR-106b-5p and enhanced by its downregulation in hippocampal neurons 50
miR-106b-5p regulates hippocampal neuronal excitability 54
IV. DISCUSSION 63
V. REFERENCES 72
VI. 국문요약 81

URI

http://dgist.dcollection.net/common/orgView/200000364452
http://hdl.handle.net/20.500.11750/16696

DOI

10.22677/thesis.200000364452

Degree

Doctor

Department

Brain and Cognitive Sciences

Publisher

DGIST

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