| Project/Area Number |
15580252
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Basic veterinary science/Basic zootechnical science
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KIMURA Kazuhiro Hokkaido Univ., Grad.School of Vet.Med., Asso.Prof., 大学院・獣医学研究科, 助教授 (30192561)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | uncoupling protein (UCP) / ATP / AMP-activated protein kinase / Knockout mice / mitochondria / glucose / ドミナントネガティブ |
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| Research Abstract |
Uncoupling proteins (UCPs) are inner mitochondrial membrane proteins that mediate dissipation of the mitochondrial membrane potential as heat rather than ATP synthesis. To clarify the cellular and molecular mechanisms of the UCPs-dependent metabolic control, especially in relation with cellular ATP levels, we firstly investigated tissue uptake of 2-deoxyglucose (2-DG) in UCP1 -knockout (KO) mice in vivo. In wild-type (WT) mice, administration of norepinephrine (NE) accelerated the disappearance of plasma 2-DG and increased 2-DG uptake into brown adipose tissue (BAT) exclusively expressing UCP1 and heart without any rise of plasma insulin level. In UCP1-KO mice, the stimulatory effect of NE on 2-DG uptake into BAT, but not into heart, disappeared completely. Insulin administration increased 2-DG uptake into BAT and also heart similarly in WT and UCP1 -KO mice. NE also increased the ratio of AMP/ATP and the activity of AMP-activated protein kinase (AMPK) in BAT of WT, but not of UCP1-KO, mice. These results suggest that the sympathetically stimulated glucose utilization in BAT is due to the serial activation of UCP1 and AMPK. We next established mammalian cells expressing functional UCP1 in a mitochondrial fraction by using Stratagene's LacSwitch II Inducible Mammalian Expression System and Hep3B human hepatocellular carcinoma that dose not express any endogenous UCP isoforms. After developing respective cells expressing functional UCP2 and UCP3, they would be useful for the studies on the mechanisms of the UCP-dependent metabolic control in an isoform-specific manner.
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