Analyses of Regulatory Mechanism of Na+-K+-Cl- Cotransporter Function in Wild Type and Mutant
| Project/Area Number |
17591939
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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 |
Functional basic dentistry
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
HIRONO Chikara Hiroshima University, Graduate School of Biomedical Sciences, Associate Professor, 大学院医歯薬学総合研究科, 助教授 (10199135)
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| Co-Investigator(Kenkyū-buntansha) |
SHIBA Yoshiki Hiroshima University, Graduate School of Bioraedical Sciences, Professor, 大学院医歯薬学総合研究科, 教授 (90110452)
SUGITA Makoto Hiroshima University, Graduate School of Biomedical Sciences, Research Associate, 大学院医歯薬学総合研究科, 助手 (50235884)
IWASA Yoshiko Hiroshima University, Graduate School of Biomedical Sciences, Research Associate, 大学院医歯薬学総合研究科, 助手 (70274090)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Na^+-K^+-2Cl^- cotransporter / salivary glands / cell culture / transfection / patch clamp / immunocytochemistry / Cl^- channel / electrolyte transport / forskolin / Calu-3細胞 / 耳下腺細胞 / 唾液腺細胞 / グラミシジン |
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| Research Abstract |
The results obtained in the research from April in 2005 to March in 2006 are as follows :
1. Establishment of culture cell system for analyses of Cl transport
Forskolin, a cAMP-increasing agent, induced an ionic current in MDCK and Calu-3 cells, which have an intrinsic Na^+-K^+-2Cl^- cotransporter activity. HEK293 cells were stained by anti-human Na^+-K^+-2Cl^- cotransporter but not by anti-rat Na^+-K^+-2Cl^- cotransporter, while the cells were stained by anti-rat Na^+-K^+-2Cl^- cotransporter after transfection with rat Na^+-K^+-2Cl^- cotransporter-containing plasmid. We started mutation experiments of the Na^+-K^+-2Cl^- cotransporter.
2. Analyses of the rat Na^+-K^+-2Cl^- cotransporter activity in salivary glands
To clarify the regulation mechanism of Na^+-K^+-2Cl^- cotransporter activity by Ca^<2+> and cAMP signals, we investigated the effects of β-adrenergic receptor stimulation on carbachol (CCh)-induced Cl^- current and the effect of α and β receptor stimulation by noradrenaline (NA).
… More
CCh induced a bumetanide-sensitive oscillatory inward Cl^- current, suggesting that the current carrier is Cl^- which moves into the cell through the Na^+-K^+-2Cl^- cotransporter. Addition of the β-adrenergic agonist, isoproterenol, and forskolin + IBMX suppressed the CCh-induced Cl^- current. This suggests that the reduction of Na^+-K^+-2Cl^- cotransporter activity by cAMP may results in decrease in electrolyte transport and then suppression of salivation during muscarinic receptor stimulation by β-adrenergic stimulation. NA induced an oscillatory anion current of which amplitude gradually decreased to a steady-state level in submandibular acinar cells. The β-adrenergic antagonist, propranolol, did not induce any current, but inhibited the gradual decrease in the NA-induced current, suggesting that the current is induced via the α-adrenergic stimulation by NA and suppressed via the β-adrenergic stimulation by NA and that the relatively small volume of saliva induced by sympathetic stimulation may be at least partially due to the reduction of Cl secretion in the acinar cells by the β-adrenergic stimulation. Less
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Report
(3 results)
Research Products
(6 results)
