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GTPγ causes contraction of skinned frog skeletal muscle via the DHP-sensitive Ca2+ channels of sealed T-tubules

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Abstract

We have investigated the involvement of G-proteins in excitation-contraction coupling of fast-twitch skeletal muscle, using a fibre preparation designed to retain intact T-tubules and sarcoplasmic reticulum. The nonhydrolysable analogue of guanosine triphosphate, GTPγS (50–500 μM) caused a strong, transient isometric contraction in this preparation. Reduction of ethylene-bis(oxonitrilo) tetraacetete (EGTA) in the sealed T-tubules from 5 mM to 0.1 mM lowered the threshold to GTPγS and removal of sodium reversibly raised it. The dihydropyridine (DHP) calcium channel antagonists nicardipine and nifedipine allowed a first contraction and then blocked subsequent GTPγS action. The phenylalkylamine methoxyverapamil (D-600) did likewise, reversibly, at 10° C. The guanosine diphosphate analogue, GDPβS, and procaine reversibly blocked the action of GTPγS pertussis toxin also blocked it. Photolytic release of 40–100 μM GTPγS within 0.1 s from S-caged GTPγS caused contraction after a latent period of 0.3–20 s. We conclude that GTPγS can activate contraction in frog skeletal muscle via a route requiring both the integrity of the T-tubular DHP-sensitive calcium channel (DHPr) and the presence of sodium in the sealed T-tubules. We propose that in this preparation GTPγS activates a G-protein, which in turn activates the DHPr as a calcium channel and releases stored calcium from within the sealed T-tubule. Implications of these results for the excitation-contraction coupling mechanism in skeletal muscle are discussed.

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Authors and Affiliations

  1. AFRC Institute of Animal Physiology, CB2 4AT, Babraham, Cambridge, UK

    B. Somasundaram & R. T. Tregear

  2. National Institute for Medical Research, Mill Hill, NW7 1AA, London, UK

    D. R. Trentham

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  1. B. Somasundaram
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  2. R. T. Tregear
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  3. D. R. Trentham
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Somasundaram, B., Tregear, R.T. & Trentham, D.R. GTPγ causes contraction of skinned frog skeletal muscle via the DHP-sensitive Ca2+ channels of sealed T-tubules. Pflügers Archiv 418, 137–143 (1991). https://doi.org/10.1007/BF00370462

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  • DOI: https://doi.org/10.1007/BF00370462

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