| Project/Area Number |
08670092
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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 |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | Hyogo College of Medicine |
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Principal Investigator |
SASAKI Sadao Hyogo Coll.Med., Fac.Med., Associate professor, 医学部, 助教授 (20104276)
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| Co-Investigator(Kenkyū-buntansha) |
HORI Kazuko Hyogo Coll.Med., Fac.Med., Assistant Professor, 医学部, 講師 (30068514)
NAKAGAKI Ikuko Hyogo Coll.Med., Fac.Med., Assistant Professor, 医学部, 講師 (90084904)
TSUJITA Junzo Hyogo Coll.Med., Fac.Med., Assistant Professor, 医学部, 講師 (30104235)
HORI Seiki Hyogo Coll.Med., Fac.Med., Professor, 医学部, 教授 (80068452)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1998: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1997: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | brown adipose tissue / cold acclimation / heat production / Ca^<2+> / CLSM / ion / uncoupling protein |
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| Research Abstract |
Brown adipose tissue cell has much quantity of Lipid droplets and motochondria which contain heat productive protein, thermogenin (uncoupling protein) and electron transfer system in the inner membrane. These are involved in the process of heat production during cold acclimation in animals.
The transport of ions in warm (WA) and cold (CA) acclimated rat brown adipocytes and the patch clamp method of reconstituted purified mitochondria of adipocytes were investigated.The electrophysiological recordings of Xenopus oocytes injected with total RNA extracted from CA and WA were also performed.
Morphometric analyses of brown adipocytes showed that lipid droplets occupied 75% of the WA cells and 37% of the CA cells. The area of a single mitochondrion in CA cells was three times that of one in the WA cells. X-ray microanalysis showed that Na was higber, and K and Cl were lower in CA than WA, in both the nucleus and the mitochondria. Moreover, Ca (total) was higher in the mitochondria, whereas was lower in the nucleus of CA cells than in WA cells. Confocal laser scanning microscopy was performed also. Acridine orange (AO), a fluorescent dye which can indicate a H^+ gradient, was localized exclusively in the mitochondria rich cytoplasm of CA cells. The fluorescence intensity of fluo-3 (Ca^<2+>indicating dye), which was loaded for 30 ruin, increased within 1 min of noradrenaline (NA) exposure in the cytoplasm of WA and CA cells. In contrast, in the nucleus with NA application fluorescence intensity rose after an initial decrease in WA, and in CA decreased gradually. The patch clamp record shows achannel activity when the reconstituted mitochondrial membrane has some voltage and pH gradient. The whole cell clamp record from Oocyte injected with total RNA of CA shows the voltage dependent outward currents.
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