Neurokinin A-Stimulated Phosphoinositide Breakdown in Rabbit Iris Sphincter Muscle
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概要
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Specific [<SUP>3</SUP>H]-substance P binding was saturable and of high affinity (K<SUB>D</SUB> = 2.5 nM) with a B<SUB>max</SUB> of 725 fmol/mg protein in the isolated rabbit iris sphincter muscle. The competition for [<SUP>3</SUP>H]-substance P binding was in the order of eledoisin > substance P > kassinin > neurokinin B > neurokinin A > physalaemin. In the same preparation, neurokinin A, as well as substance P induced a concentration-related accumulation of [<SUP>3</SUP>H]-inositol phosphates (IPs), and the maximum increase was about 200% of the control at 10<SUP>-4</SUP> M. [D-Arg<SUP>1</SUP>, D-Trp<SUP>7, 9</SUP>, Leu<SUP>11</SUP>]-substance P (SP) and [D-Pro<SUP>2</SUP>, D-Trp<SUP>7, 9</SUP>]-SP (10<SUP>-3</SUP> M) inhibited substance P or neurokinin A (10<SUP>-4</SUP> M)-induced phosphatidylinositol 4, 5-bisphosphate (PIP<SUB>2</SUB>) hydrolysis significantly. [D-Arg<SUP>1</SUP>, D-Pro<SUP>2</SUP>, D-Trp<SUP>7, 9</SUP>, Leu<SUP>11</SUP>]-SP (10<SUP>-3</SUP> M) also inhibited neurokinin A (10<SUP>-4</SUP> M)-induced PIP<SUB>2</SUB> hydrolysis significantly. Neurokinin A and substance P produced concentration-related contractions in normal Ca<SUP>2+</SUP>-containing medium. The contractile response was weaker in Ca<SUP>2+</SUP>-free medium, and there was no response in 0.2 mM EGTA medium. In Ca<SUP>2+</SUP>-free medium, the basal level of [<SUP>3</SUP>H]-IPs accumulation was smaller than that in normal medium, and neurokinin A and substance P significantly increased PIP2 hydrolysis. In the 0.2 mM EGTA containing medium, neurokinin A and substance P did not stimulate the PIP<SUB>2</SUB> hydrolysis. These results suggest that in the rabbit iris sphincter muscle, there are tachykinin receptors linking to PIP<SUB>2</SUB> hydorolysis and Ca<SUP>2+</SUP> mobilization and that these mechanisms underlie the mechanism for the neurokinin A-induced contractile response, as well as the substance P-induced one.
著者
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Fujiwara Motohatsu
Department Of Pharmacology Faculty Of Medicine Kyoto University
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Yamamoto Masato
Department Of Cadiovascular Surgery Tokyo Women's Medical University Medical Center East
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Ninomiya Haruaki
Department Of Biological Regulation Faculty Of Medicine Tottori University
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TANIGUCHI Takashi
Department of Computational Science and Engineering, Nagoya University
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Ninomiya Haruaki
Department of Pharmacology, Faculty of Medicine, Kyoto University
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Fukunaga Reiko
Department of Neurobiology, Kyoto Pharmaceutical University
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Ebii Koichi
Department of Neurobiology, Kyoto Pharmaceutical University
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Yamamoto Masato
Department of Neurobiology, Kyoto Pharmaceutical University
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Taniguchi Takashi
Department of Biology, Kyoto Pharmaceutical University
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