Effects of the PKA inhibitor H-89 on excitation-contraction coupling in skinned and intact skeletal muscle fibres

R. Blazev, M. Hussain, Tony Bakker, S.I. Head, G.D. Lamb

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    16 Citations (Scopus)


    This study investigated the effects of the protein kinase A (PKA) inhibitor, H-89, in mechanically-skinned muscle fibres and intact muscle fibres, in order to determine whether PKA phosphorylation is essential for normal excitation-contraction (E-C) coupling. In skinned EDL fibres of the rat, force responses to depolarization (by ion substitution) were inhibited only slightly by 10 muM H-89, a concentration more than sufficient to fully inhibit PKA. Staurosporine (1 muM), a potent non-specific kinase inhibitor, also had little if any effect on depolarization-induced responses. At 1-2 muM, H-89 significantly slowed the repriming rate in rat skinned fibres, most likely due to it deleteriously affecting the T-system potential. With 100 muM H-89, the force response to depolarization by ion substitution was completely abolished. This inhibitory effect was reversed by washout of H-89 and was not due to block of the Ca2+ release channel in the sarcoplasmic reticulum (SR). In intact single fibres of the flexor digitorum longus (FDB) muscle of the mouse, 1-3 muM H-89 had no noticeable effect on action-potential-mediated Ca2+ transients. Higher concentrations (4-10 muM) caused Ca-2+ transient failure in fibres stimulated at 20 Hz in a manner indicative of action-potential failure. At 10-100 muM, H-89 also inhibited net Ca2+ uptake by the SR and affected the Ca2+-sensitivity of the contractile apparatus in rat skinned fibres. All such effects were proportionately greater in toad muscle fibres. These results do not support the hypothesis that phosphorylation is essential for the Ca2+ release channel to open in response to voltage-sensor activation in skeletal muscle fibres.
    Original languageEnglish
    Pages (from-to)277-286
    JournalJournal of Muscle Research and Cell Motility
    Publication statusPublished - 2001


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