| Project/Area Number |
11670038
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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 |
General physiology
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| Research Institution | Nagoya University |
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Principal Investigator |
NAKAYAMA Shinsuke Sch.of Med., Nagoya University, Assoc. Prof., 医学部, 助教授 (30192230)
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| Co-Investigator(Kenkyū-buntansha) |
OKAMURA Yasushi Natl.Inst.Advanced Industrial Science & Technology, Agency of Industrial Science and Technology, Senior Researcher, 生体分子工学部, 主任研究官
IWASHITA Toshihide Sch.of Med., Nagoya University, Assistant Professor, 医学部, 講師 (00283432)
KUZUYA Masafumi Sch.of Med., Nagoya University, Assistant Professor, 医学部, 講師 (10283441)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Calcium / Ion channels / Electrophysiology / Molecular biology / Inactivation / Persistent current / Multiple open states |
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| Research Abstract |
In smooth muscle, it is well known that spike activities are largely due to activation of voltage-sensitive Ca^<2+> channels. Sustained Ca^<2+> influx seen during long term depolarization of the cell membrane is also considered to be through these voltage-sensitive Ca^<2+> channels. Since both phenomena are significantly suppressed by Ca^<2+> antagonists, (dihydropyridinesensitive) L-type Ca^<2+> channels seem to cover a wide range of smooth muscle behavior : from transient to persistent events.
Previously, we have demonstrated the presence of multiple open states in smooth muscle L-type Ca^<2+> channels, in guinea-pig urinary bladder, taenia caeci and gastric antrum. During a large, long duration depolarization, the conformation of Ca^<2+> channels are transferred to the second open state in which these channels do not, or very slowly inactivate during depolarizaton, and deactivate very slowly upon repolarization.
In 1999, using cell-attached patch clamp techniques, we mainly examined w
… More
hether smooth muscle Ca^<2+> channel undergoes voltage-dependent conversion of the channel conformation from normal to the second open state, even when its α_1-subunits are expressed in the plasma membrane. The answer was 'YES'. We demonstrated that α_1-subunit of cloned smooth muscle Ca^<2+> channel (α_<1C-b>) does not inactivate during large conditioning depolarization, and produced slow deactivating tail current upon repolarization of the cell membrane. These phenomena were essentially the same observed in intact smooth muscle Ca^<2+> channels.
In 2000, we used whole-cell patch clamp techniques to investigate 1) the role of β-subunit in the voltage-dependent conversion of the C^<2+> channel conformation, and 2) interaction of depolarization and Ca^<2+> agonist. The experiments revealed that a smooth muscle β-subunit (β_3) decreased the ratio of the Ca^<2+> channels converted to the second open state during large depolarization, and that voltage-dependent conversion and Ca^<2+> agonistinduced mode 2 gating mechanism operated separately, causing four open states in α_1-subunit of cloned smooth muscle Ca^<2+> channel. Less
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