서강대학교

목차

TABLE OF CONTENTS ⅳ
LIST OF TABLES ⅶ
LIST OF FIGURES ⅶ


ABSTRACT 1
BACKGROUNDS 3
Adipose tissue 3
Non-shivering thermogenesis 4
Beiging and Browning of WAT 4
Long-chain acyl-CoA synthetase 1 (ACSL1) 5
Mitochondrial phospholipid remodeling, cardiolipin (CL) maturation 6
Objective 6

MATERIAL AND METHODS
Animals 8
Cell Culture 8
Cold Exposure 9
Western Blot 9
Quantitative qRT-PCR 10
Pre-Adipocyte Isolation 10
Differentiation of Adipocytes and Cold Stimuli 10
FACS Analysis 11
Radiolabeled Palmitate Solution 12
Fatty Acid Oxidation (FAO) Assay 12
Re-esterification Assay 13
Organelle Fractionation 14
Lipid Extraction from Tissues 15
Mitochondria Lipid TLC 16
Lipidomics Analysis 16
Cell Transfection and Lentivirus Infection 17

RESULTS
Acsl1 and other isoforms expression level in RT and cold condition 20
ACSL1 subcellular localization during cold exposure 25
Loss of ACSL1 in BAT leads to decreased level of lipids accompanied by cold intolerance 28
BAKO mice didn’t show thermogenic gene induction during acute cold exposure 31
BAKO mice iWAT shows UCP1 compensation 34
ACSL1 may function differently in iWAT and BAT 37
Loss of ACSL1 leads to decreased PKA signaling 40
Mitochondria quality and function are both impaired 43
ACSL1 seems to regulate thermogenesis from mitochondria homeostasis fundamentally 46
Loss of ACSL1 leads to mitochondrial abnormalities 49
Altered cardiolipin composition of mitochondria underlie cold intolerance 52
Broad profiling of lipid species alterations in the absence of ACSL1 (ESI + mode) 55
Broad profiling of lipid species alterations in the absence of ACSL1 (ESI - mode) 58
Cardiolipin remodeling is impaired in ACSL1-deficient BAT mitochondria 61
Impaired fission could be the result of swollen mitochondria 64

DISSCUSION 67
REFERENCE 71