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Effects of a phorbol ester on acetylcholine‐induced Ca2+ mobilization and contraction in the porcine coronary artery.
Author(s) -
Itoh T,
Kubota Y,
Kuriyama H
Publication year - 1988
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.1988.sp017008
Subject(s) - ionomycin , phosphatidylinositol , acetylcholine , phorbol , chemistry , contraction (grammar) , phosphatidic acid , extracellular , activator (genetics) , muscle contraction , protein kinase c , medicine , endocrinology , biophysics , fura 2 , biochemistry , intracellular , biology , kinase , cytosol , phospholipid , enzyme , receptor , membrane
1. The effects of 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), an activator of protein kinase C, have been investigated on intact and chemically skinned muscle strips of the porcine coronary artery. 2. In the presence or absence of extracellular Ca2+, TPA (0.1‐1 nM) slightly enhanced the amplitude of ACh (10 microM)‐induced contractions but at 100 nM, inhibited the contractions by approximately 50%. 3. ACh (10 microM) reduced the amount of [32P]phosphatidylinositol 4,5‐bisphosphate (PIP2) and increased the amount of [32P]phosphatidic acid (PA) in the presence or absence of Ca2+. TPA (over 1 nM) dose‐dependently inhibited the hydrolysis of PIP2 induced by ACh. 4. ACh (over 0.1 microM) dose‐dependently increased the intensity of fura‐2 fluorescence in dispersed single‐cell suspensions. TPA (over 1 nM) dose‐dependently inhibited the increase of the Ca2+ transient evoked by ACh, but it did not modify the ionomycin‐induced Ca2+ transient or the resting fluorescence. These inhibitory effects of TPA occurred over a similar dose range to that which inhibited ACh‐induced PIP2 break‐down. 5. When the relationship between ACh‐induced contraction amplitude and Ca2+ transient was observed in the presence or absence of 10 nM‐TPA, TPA greatly reduced the Ca2+ transient but did not modify the amplitude of contraction. 6. In saponin‐treated skinned muscle strips, TPA (10 nM) or 1,2‐diolein (50 micrograms/ml) with phosphatidylserine (PS; 50 micrograms/ml) increased the amplitude of contraction evoked by various concentrations of Ca2+ (0.1‐1.0 microM) without any change in the maximum amplitude of the Ca2+‐induced contraction. 7. TPA (10 nM) with PS (50 micrograms/ml) increased the amplitude of contraction evoked by 10 microM‐inositol 1,4,5‐trisphosphate in chemically skinned muscle strips. 8. It is concluded that TPA inhibits the ACh‐induced [Ca2+]i increase by inhibiting the hydrolysis of PIP2, but that it enhances the Ca2+ sensitivity of the contractile proteins. These results indicate that ACh‐induced contractions are controlled by negative feed‐back regulation of PIP2 hydrolysis together with a positive feed‐back regulation of the Ca2+ sensitivity of the contractile proteins. This may depend on the on‐going level of protein kinase C activation.