Summary
Depolarization-induced Ca2+ release was studied in a mixture of triads and terminal cisternae isolated from rabbit skeletal muscle. The vesicles were actively loaded with known amounts of Ca2+ in the absence of precipitating anions in a solution containing 100 mm K propionate buffer. Changes in extravesicular Ca2+ were monitored with 10 μ m Fura-2 (membrane impermeant form). Ca2+ release was initiated by diluting an aliquot of the loaded vesicles into a TEACl release solution designed to maintain a constant [K+] · [Cl−] product. Fast release, defined as the percentage of total Ca2+ loaded which released in less than 10 sec, occurred when extravesicular free Ca2+ was in the submicromolar range and was unaffected by 5 mm caffeine under depolarizing conditions, change in external pH to 6.5, and an increase in external Mg2+ concentration from 0.1 to 0.2 mm. Thus, the Ca2+ release measured in these studies is distinct from Ca2+-induced Ca2+ release. The fast release more than doubled when a greater dilution (1 ∶ 20 versus 1 ∶ 10) of the loaded vesicles into the release solution, which would produce a larger depolarization, was used. The percentage of loaded Ca2+ which released rapidly in a particular triad preparation was similar to the percentage of vesicles structurally coupled as visualized by electron microscopy.
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We thank Gerry Vaio and Melanie Vander Klok for excellent technical support. This work was supported by the National Institutes of Health program project grant PO1-HL27867, NSF Biological Instrumentation Grant DIR-8812094 and State of Ohio Research Challenge Grant.
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Corbett, A.M., Bian, J., Wade, J.B. et al. Depolarization-induced calcium release from isolated triads measured with impermeant Fura-2. J. Membarin Biol. 128, 165–179 (1992). https://doi.org/10.1007/BF00231810
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DOI: https://doi.org/10.1007/BF00231810