1996 Fiscal Year Final Research Report Summary
Molecular analysis of heart inwardly rectifying K+ channels
Project/Area Number |
07670779
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Circulatory organs internal medicine
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Research Institution | Osaka University |
Principal Investigator |
KURODA Masao Faculty of Medicine, Osaka University, Lecturer, 医学部, 講師 (90028556)
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Co-Investigator(Kenkyū-buntansha) |
ISOMOTO Shojiro Faculty of Medicine, Osaka University, Assistant Professor, 医学部, 助手 (80273671)
HORIO Yoshiyuki Faculty of Medicine, Osaka University, Lecturer, 医学部, 講師 (30181530)
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Project Period (FY) |
1995 – 1996
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Keywords | inwardly rectifying K+ channels / heart / ATP-sensitive inwardly rectifying K+ channels / RT-PGR / cloning / iK1 channel / sulphonylurea receptor / 培養細胞 |
Research Abstract |
In myocytes, inwardly rectifying K+ channels have been considered to contribute decreasing membrane potential, and maintaining long action potential in depolalization of the cells. To investigate molecular study of cardiac inwardly rectifying K+ channels, using oocyte expression system, we co-expressed IRK1/Kir2.1 and IRK2/Kir2.2 or expressed a fused cDNA of IRK1 and IRK2, which were both found to be expressed in the heart. We found that IRK1 and IRK2 were expressed independently. We performed RT-PCR experiment with a single cell of atrial myocyte or ventricular myocyte to detect expression of IRK1 and IRK2. The experiment showed that IRK2 was expressed both cardiac myocytes but IRK1 was not expressed. Thus iK1 channel in the heart was considered to be IRK2. We also studied molecular cloning of heart ATP-sensitive inwardly rectifying K+ channels. We cloned BIR and uKATP-1 as a channel subunits and SUR2A and SUR2B as a sulfonylurea receptor subunits. We expressed these clones in HEK293T cells and assayd with patch-clamp method. Using these clones we found that heart ATP-sensitive inwardly rectifying K+ channels were composed of BIR and SUR2B.
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[Publications] Takumi T,Ishii T,Horio Y,Morishige K,Takahashi N,Yamada M,Yamashita T,Kiyama H,Sohmiya K,Nakanishi S,Kurachi Y: "A novel ATP-dependent inward rectifier potassium channel expressed predominantly in glial cells." The Journal of Biological Chemistry. 270. 16339-16346 (1995)
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[Publications] Ito M,Inanobe A,Horio Y,Hibino H,Isomoto S,Ito H,Mori K,Tonosaki A,Tomoike H,Kurachi Y: "Immunolocalization of an inwardly rectifying K+ channel, K_<AB>-2 (Kir4.1), in the basolateral membrane of renal distal tubular epithelia." FEBS Letters. 388. 11-15 (1996)
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[Publications] Horio Y,Hibino H,Inanobe A,Yamada M,Ishii M,Tada Y,Sato E,Hata Y.Takai Y,Kurachi Y: "Clustering and enhanced activity of an inwardly rectifying potassium channel, Kir4.1.by an anchoring protein, PSD-95/SAP 90 family." The Journal of Biological Chemistry. (in press).
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[Publications] Hibino H,Horio Y,Inanobe A,Doi K,Tamada M,Gotow T,Uchiyama Y,Kubo T,Kurachi Y: "An ATP-dependent inwardly rectifying potassium channel, K_<AB>-2 (Kir4.1), in cochlear stria vascularis of inner ear : its specific subcellular localization and correlation with the formation of endocochlear potential." The Journal of Neuroscience. (in press).
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