Title

Comparative metabolomic analysis of corticosterone-induced changes in embryonic mouse hypothalamic cells using gas chromatography coupled with triple-quadrupole tandem mass spectrometry

Alternative Title

GC-QqQ-MS/MS를 사용한 쥐 배아 시상 하부 세포에서의 코르티코스테론 유도 대사체 변화 분석

DGIST Authors

Jiwoo Yeom ; Eun-Kyoung Kim ; Myungjin Kim

Advisor

김은경

Co-Advisor(s)

Myungjin Kim

Issued Date

2021

Awarded Date

2021/02

Citation

Jiwoo Yeom. (2021). Comparative metabolomic analysis of corticosterone-induced changes in embryonic mouse hypothalamic cells using gas chromatography coupled with triple-quadrupole tandem mass spectrometry. doi: 10.22677/thesis.200000361606

Type

Thesis

Subject

hypothalamic cells, corticosterone, neuropeptides, metabolomics, 시상 하부 세포, 코르티코스테론, 신경 펩티드, 대사체학

Table Of Contents

I. Introduction 1
II. Materials and methods 6
1. Cell culture 6
2. Cell viability assay 6
3. Real time-PCR 7
4. Metabolomics analysis by GC-QqQ-MS/MS 8
4.1 Glycolysis, TCA cycle, and PPP intermediates analysis 8
4.2 Amino acids analysis 9
4.3 Free fatty acids analysis 10
5. ATP assay 11
III. Results 12
1. Corticosterone has cytotoxicity in embryonic mouse hypothalamic cells in a dose-dependent manner 12
2. Corticosterone increases mRNA expression of Agrp, Npy, and Pomc in embryonic mouse hypothalamic cells 12
3. Corticosterone changes metabolites in different ways depending on the cell type 14
3.1 Corticosterone increases glucose metabolism intermediates and decreases amino acids and fatty acids in N41 cells 14
3.2 Corticosterone decreases glucose metabolism intermediates and increases fatty acids in N43/5 cells 16
4. Corticosterone decreases intracellular ATP level and regulates mRNA expression related to fatty acid metabolism in N41 cells 16
5. Pharmacological inhibition of GPAT blocks the increase of Npy and Gpat mRNA expression induced by corticosterone in N41 cells 18
6. Corticosterone decreases intracellular ATP level and increases mRNA expression of Gpat in N43/5 cells 18
7. Pharmacological inhibition of GPAT blocks increase of Pomc mRNA expression induced by corticosterone in N43/5 cells 21
IV. Discussion 24
V. Conclusion 27
VI. References 28

URI

http://dgist.dcollection.net/common/orgView/200000361606
http://hdl.handle.net/20.500.11750/16640

DOI

10.22677/thesis.200000361606

Degree

Master

Department

Brain and Cognitive Sciences

Publisher

DGIST

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