1990 Fiscal Year Annual Research Report

心筋イオンチャンネル

Research Project

Project/Area Number	02257107
Research Institution	Juntendo University
Principal Investigator	大地陸男順天堂大学, 医学部, 教授 (10049025)
Co-Investigator(Kenkyū-buntansha)	三品昌美新潟大学, 脳研, 教授 (80144351) 栗山煕九州大学, 医学部, 教授 (40037495) 平則夫東北大学, 医学部, 教授 (60004553) 亀山正樹広島大学, 医学部, 助教授 (60150059) 野間昭典九州大学, 医学部, 教授 (00132738)
Keywords	カルシウムチャンネル / カリウムチャンネル / ATP / カルパスタチン / 伸展感受性チャンネル / 心筋 / 血管平滑筋 / イオンチャンネル
Research Abstract	心臓の電気活動の電気的素過程、特にイオンチャンネル開閉とその分子的機序を解明し、心臓死の主因をなす不整脈の制御に貢献することが我々の研究目的であった。Caチャンネルについてはパッチクランプ法で単一チャンネル電流が解析され、イソプレナリンがチャンネルの燐酸化により利用率の増大をもたらし、心筋の代謝産物であるアデノシンが、この増大を抑制することが判明した。また、Caチャンネルのrundownを防止する内因性蛋白の同定がクロマトグラフ法とパッチクランプ法で進められ、カルパスタチンと酷似することが判明した。さらに、細胞内Ca濃度がCaチャンネルの制御に影響することがIndoー1を用いてほぼ確認された。リガンド結合実験も行われ、イオン化するジヒドロピリジン(DHP)Ca拮抗薬NKYー722が心筋細胞膜でDHP受容体以外にも結合することが示唆された。Kチャンネルについては、Naーactivated Kチャンネルのコンダクタンスの性質が解明され、細胞内NaやMgによるブロックが判明し、ATPー感受性Kチャンネルについてもニコランジルなどの薬物による修飾も解明された。伸張感受性チャンネルについては、非筋性細胞で詳細な解析を行ったが、心筋にも存在するらしいので同じ手法で研究を行っている。血管平滑筋にいては、門脈平滑筋にATP受容器作動性チャンネルルとATP感受性Kチャンネルが存在した。さらに冠動脈平滑筋にもATP感受性Kチャンルの存在が判明したのでこの研究を行う。チャンネル蛋白の構造機能相関については、ニコチン性アセチルコリン受容体のアミノ基に点変異を導入し、カチオンの選択性に影響する部位を決定した。心筋メッセンジャ-RNAの卵母細胞注入によるチャンネル機能発現の研究が計画している。

Research Products

(37 results)

All Other

All Publications (37 results)

[Publications] R.Ochi: "Modulation of slow gating process of calcium channels by isoprenaline in guineaーpig ventricular cells" Journal of Physiology. 424. 187-204 (1990)
[Publications] M.Kato: "Mechanism of adenosineーinduced inhibition of calcium channels by isoprenaline in guinea pig ventricular cells" Circulation Research. 67. 1134-1141 (1990)
[Publications] M.Kameyama: "Intracellular mechanisms involved in “runーdown" of calcium channels" Advance of Experimental Medicine and Biology. 255. 111-118 (1990)
[Publications] K.Yazawa: "Mechanism of receptorーmediated regulation of the delayed outward potassium current in guineaーpig ventricular myocytes" Journal of Physiology. 421. 135-150 (1990)
[Publications] K.Yazawa: "An improved method for isolating cardiac myocytes useful for patch clamp studies" Japanese Journal of Physiology. 40. 157-163 (1990)
[Publications] N.Tohse: "Protein kinase C activation enhances the delayed rectifier potassium current in guinea pig heart" Journal of Molecular and Cellular Cardiology. 22. 725-738 (1990)
[Publications] K.Tsuji: "The effect of structural modification of several groups on the Dーring of grayanotoxin on its depolarization potency in squid giant axon" The journal of Pharmacology and Experimental Therapeutics.
[Publications] Z.Wang: "Conductance properties of the Na^+ーactivated K^+ channel in guineaーpig ventricular cells" Journal of Physiology.
[Publications] F.M.Tareen: "βーAdrenergic and muscarinic regulation of the chloride current in guineaーpig ventricular cells" Journal of Physiology.
[Publications] D.Qin: "Limitations due to unstirred layers in measuring channel response of excised membrane patch using rapid solution exchange method" Japanese Journal of Physiology.
[Publications] M.Takano: "Selective modulation of the ATPーsensitive K^+ channel by nicorandil in guineaーpig cardiac cell membrane" NaunynーSchmiedeberg's Archiv of Pharmacology. 342. 592-597 (1990)
[Publications] M.Sokabe: "Electrophysiological analysis of structural aspects of voltageーdependent SR K channel" Comparative Biochemistry.
[Publications] M.Sokabe: "Quantitative videomicroscopy of patch clamped membrames:stress,strain,capacitance,and stretch channel activation" Biophysical Journal.
[Publications] M.Sokabe: "The structure and dynamics of patch clamped membranes:A study by using differential interference light microscopy" Journal of Cell Biology. 111. 599-606 (1990)
[Publications] K.Nomura: "Aminoglycoside blockade of Ca activated K channel from rat brain synaptosomal membranes incorporated into planar bilayers." Journal of Membrane Biology. 115. 241-251 (1990)
[Publications] F.Sachs: "Stretch activated ion channels and membrane mechanics" Neuroscience Research. 12. S1-4 (1990)
[Publications] F.Ito: "Effects of ions and drugs on the responses of sensory axon terminals of decapsulated frog muscle spindles" Neuroscience Research. 12. S15-S26 (1990)
[Publications] K.Kitamura: "Similarity and diversity of electrical activity in vascular smooth muscles" Progress in Clinical and Biological Research. 327. 257-276 (1990)
[Publications] K.Okabe: "Actions of cromakalim on ionic currents recorded from single smooth muscle cells of the rat portal vein" The Journal of Pharmacology and Experimental Therapeutics. 252. 832-839 (1990)
[Publications] Y.Inoue: "Endothelin augments unitary calcium chanel currents on the smooth muscle cell membrane of guineaーpig portal vein" Journal of Physiology. 423. 171-191 (1990)
[Publications] Z.Xiong: "ATP activates cationic currents and moduates the Ca current through GTPーbinding protein in rabbit portal vein" Journal of Physiology.
[Publications] Z.Xiong: "Pinacidil inhibits the ryanodineーsensitive outward current and glibenclamide antagonises its action in cells from the rabbit portal vein" British Journal of Pharmacology.
[Publications] M.B.Jackson: "Spontaneous and agonistーinduced openings of an acetylcholine receptor channel composed of bovine muscle αー,βー and δーsubunits" Pflugers Archiv. 417. 129-135 (1990)
[Publications] K.Sakimura: "Functional expresssion from cloned cDNAs of glutamate receptor species responsive to kainate and quisqualate" FEBS Letters. 272. 73-80 (1990)
[Publications] M.Mishina: "Structure and function of acetylcholine and glutamate receptors" Fidia Research Foundation Symposium.
[Publications] T.Konno: "Structural determinants of ion selectivity in the acetylcholine receptor channel" Proceedings of the Royal Society of London B.
[Publications] K.Nunoki: "Specific binding sites of NKYー722,a hydrophobic 1,4 dihydropyridine derivative (DHP) in the canine microsomal fractions: comparison with PN 200ー110 binding" Japanese Journal of Pharmacology. 52(Sup). 331 (1990)
[Publications] N.Nunoki: "Effects of SDー3211,a benzothiazine Ca^<2+> channel blocker on dihydropyridine (DHP) binding to the canine cardiac membrane" Japanese Journal of Physiology. 55(Sup). 91 (1991)
[Publications] R.Ochi: "Increase in modeー2 sweeps by isoprenaline in cardiac calcium channel treated with BAY K 8644" Japanese Journal of Physiology. 40(Sup). S261 (1990)
[Publications] R.Ochi: "Modulation of gating process of calcium channel by isoproterenol and nitrendipine in cardiac muscle" Recent Advances in Calcium Channels and Calcium Antagonists.eds.K.Yamada & S.Shibata,Pergamon Press,New York. 11-17 (1990)
[Publications] M.Kamayama: "Mechanisms of rundown of cardiac calcium channels" Recent Advances in Calcium Channels and Calcium Antagonists.eds.K.Yamada & S.Shibata,Pergamon Press,New York. 3-10 (1990)
[Publications] A.Noma: "The cardiac ATPーsensitive K^+ channel" Ionic Currents and Ischemia.eds.J.Vereecke,P.P.Bogaert & Verdonck,Leuven University Press,Belgium. 99-119 (1990)
[Publications] M.Sokabe: "Towards molecular mechanism of the gating in stretch activated channels" Comparative Aspect of Mechanoreception,ed.F.Ito,Springer Verlag.
[Publications] K.Kitamura: "Characteristics of vascular smooth muscle cell membranes and their modifying factors" Coronary Circulation in Physiological and Pathophysiological States,eds.M.Nakamura & P.M.Vanhoutte,Springer Verlag,Tokyo. 125-156 (1990)
[Publications] 三品昌美: "アセチルコリン受容体の構造と機能" 神経研究の進歩. 34. 911-919 (1990)
[Publications] 崎村建司: "グルタミン酸受容体チャンネルの構造と機能" 実験医学.
[Publications] 三品昌美: "分子生物科学第10巻,神経情報の伝達" 岩波書店, (1991)

1990 Fiscal Year Annual Research Report

心筋イオンチャンネル

Principal Investigator

大地 陸男 順天堂大学, 医学部, 教授 (10049025)

Research Products

[Publications] R.Ochi: "Modulation of slow gating process of calcium channels by isoprenaline in guineaーpig ventricular cells" Journal of Physiology. 424. 187-204 (1990)

[Publications] M.Kato: "Mechanism of adenosineーinduced inhibition of calcium channels by isoprenaline in guinea pig ventricular cells" Circulation Research. 67. 1134-1141 (1990)

[Publications] M.Kameyama: "Intracellular mechanisms involved in “runーdown" of calcium channels" Advance of Experimental Medicine and Biology. 255. 111-118 (1990)

[Publications] K.Yazawa: "Mechanism of receptorーmediated regulation of the delayed outward potassium current in guineaーpig ventricular myocytes" Journal of Physiology. 421. 135-150 (1990)

[Publications] K.Yazawa: "An improved method for isolating cardiac myocytes useful for patch clamp studies" Japanese Journal of Physiology. 40. 157-163 (1990)

[Publications] N.Tohse: "Protein kinase C activation enhances the delayed rectifier potassium current in guinea pig heart" Journal of Molecular and Cellular Cardiology. 22. 725-738 (1990)

[Publications] K.Tsuji: "The effect of structural modification of several groups on the Dーring of grayanotoxin on its depolarization potency in squid giant axon" The journal of Pharmacology and Experimental Therapeutics.

[Publications] Z.Wang: "Conductance properties of the Na^+ーactivated K^+ channel in guineaーpig ventricular cells" Journal of Physiology.

[Publications] F.M.Tareen: "βーAdrenergic and muscarinic regulation of the chloride current in guineaーpig ventricular cells" Journal of Physiology.

[Publications] D.Qin: "Limitations due to unstirred layers in measuring channel response of excised membrane patch using rapid solution exchange method" Japanese Journal of Physiology.

[Publications] M.Takano: "Selective modulation of the ATPーsensitive K^+ channel by nicorandil in guineaーpig cardiac cell membrane" NaunynーSchmiedeberg's Archiv of Pharmacology. 342. 592-597 (1990)

[Publications] M.Sokabe: "Electrophysiological analysis of structural aspects of voltageーdependent SR K channel" Comparative Biochemistry.

[Publications] M.Sokabe: "Quantitative videomicroscopy of patch clamped membrames:stress,strain,capacitance,and stretch channel activation" Biophysical Journal.

[Publications] M.Sokabe: "The structure and dynamics of patch clamped membranes:A study by using differential interference light microscopy" Journal of Cell Biology. 111. 599-606 (1990)

[Publications] K.Nomura: "Aminoglycoside blockade of Ca activated K channel from rat brain synaptosomal membranes incorporated into planar bilayers." Journal of Membrane Biology. 115. 241-251 (1990)

[Publications] F.Sachs: "Stretch activated ion channels and membrane mechanics" Neuroscience Research. 12. S1-4 (1990)

[Publications] F.Ito: "Effects of ions and drugs on the responses of sensory axon terminals of decapsulated frog muscle spindles" Neuroscience Research. 12. S15-S26 (1990)

[Publications] K.Kitamura: "Similarity and diversity of electrical activity in vascular smooth muscles" Progress in Clinical and Biological Research. 327. 257-276 (1990)

[Publications] K.Okabe: "Actions of cromakalim on ionic currents recorded from single smooth muscle cells of the rat portal vein" The Journal of Pharmacology and Experimental Therapeutics. 252. 832-839 (1990)

[Publications] Y.Inoue: "Endothelin augments unitary calcium chanel currents on the smooth muscle cell membrane of guineaーpig portal vein" Journal of Physiology. 423. 171-191 (1990)

[Publications] Z.Xiong: "ATP activates cationic currents and moduates the Ca current through GTPーbinding protein in rabbit portal vein" Journal of Physiology.

[Publications] Z.Xiong: "Pinacidil inhibits the ryanodineーsensitive outward current and glibenclamide antagonises its action in cells from the rabbit portal vein" British Journal of Pharmacology.

[Publications] M.B.Jackson: "Spontaneous and agonistーinduced openings of an acetylcholine receptor channel composed of bovine muscle αー,βー and δーsubunits" Pflugers Archiv. 417. 129-135 (1990)

[Publications] K.Sakimura: "Functional expresssion from cloned cDNAs of glutamate receptor species responsive to kainate and quisqualate" FEBS Letters. 272. 73-80 (1990)

[Publications] M.Mishina: "Structure and function of acetylcholine and glutamate receptors" Fidia Research Foundation Symposium.

[Publications] T.Konno: "Structural determinants of ion selectivity in the acetylcholine receptor channel" Proceedings of the Royal Society of London B.

[Publications] K.Nunoki: "Specific binding sites of NKYー722,a hydrophobic 1,4 dihydropyridine derivative (DHP) in the canine microsomal fractions: comparison with PN 200ー110 binding" Japanese Journal of Pharmacology. 52(Sup). 331 (1990)

[Publications] N.Nunoki: "Effects of SDー3211,a benzothiazine Ca^<2+> channel blocker on dihydropyridine (DHP) binding to the canine cardiac membrane" Japanese Journal of Physiology. 55(Sup). 91 (1991)

[Publications] R.Ochi: "Increase in modeー2 sweeps by isoprenaline in cardiac calcium channel treated with BAY K 8644" Japanese Journal of Physiology. 40(Sup). S261 (1990)

[Publications] R.Ochi: "Modulation of gating process of calcium channel by isoproterenol and nitrendipine in cardiac muscle" Recent Advances in Calcium Channels and Calcium Antagonists.eds.K.Yamada & S.Shibata,Pergamon Press,New York. 11-17 (1990)

[Publications] M.Kamayama: "Mechanisms of rundown of cardiac calcium channels" Recent Advances in Calcium Channels and Calcium Antagonists.eds.K.Yamada & S.Shibata,Pergamon Press,New York. 3-10 (1990)

[Publications] A.Noma: "The cardiac ATPーsensitive K^+ channel" Ionic Currents and Ischemia.eds.J.Vereecke,P.P.Bogaert & Verdonck,Leuven University Press,Belgium. 99-119 (1990)

[Publications] M.Sokabe: "Towards molecular mechanism of the gating in stretch activated channels" Comparative Aspect of Mechanoreception,ed.F.Ito,Springer Verlag.

[Publications] K.Kitamura: "Characteristics of vascular smooth muscle cell membranes and their modifying factors" Coronary Circulation in Physiological and Pathophysiological States,eds.M.Nakamura & P.M.Vanhoutte,Springer Verlag,Tokyo. 125-156 (1990)

[Publications] 三品 昌美: "アセチルコリン受容体の構造と機能" 神経研究の進歩. 34. 911-919 (1990)

[Publications] 崎村 建司: "グルタミン酸受容体チャンネルの構造と機能" 実験医学.

[Publications] 三品 昌美: "分子生物科学 第10巻,神経情報の伝達" 岩波書店, (1991)

大地陸男順天堂大学, 医学部, 教授 (10049025)

[Publications] 三品昌美: "アセチルコリン受容体の構造と機能" 神経研究の進歩. 34. 911-919 (1990)

[Publications] 崎村建司: "グルタミン酸受容体チャンネルの構造と機能" 実験医学.

[Publications] 三品昌美: "分子生物科学第10巻,神経情報の伝達" 岩波書店, (1991)