Ca2+ loading reduces the tensile strength of sarcolemmal vesicles shed from rabbit muscle

1. Sarcolemmal vesicles shed by rabbit muscle were loaded with Ca2+ by means of A23187 or ionomycin. [Ca2+]0 was buffered between 0.8 and 20 microM. Membrane strength was measured by pipette aspiration. 2. At 20 microM Ca2+ many vesicles underwent autolysis, or were so weak that they burst instantly on aspiration. Between 10 and 2 microM Ca2+ a graded decrease in membrane strength was demonstrable. At 0.8 microM Ca2+ the mechanical properties of the sarcolemma remained unaltered. 3. Mg2+ carried by A23187 does not mimic the effect of Ca2+. The ionophore itself similarly did not cause a decrease in membrane tensile strength. 4. Pre-treatment with BAPTA-AM, so as to buffer internal Ca2+, partly protected vesicles against the decrease in membrane strength produced by Ca2+ loading. 5. Membrane strength was not restored by adding excess BAPTA to the bathing solution, so as to reverse the Ca2+ gradient. An irreversible degradation of the membrane consequent upon raised [Ca2+]1 seems indicated. 6. These findings are discussed in relation to the mechanisms which have been advanced to account for the role of elevated [Ca2+]1 in cell death. 7. An attempt to use staphylococcal alpha-toxin as an alternative means to permeabilize the sarcolemma led to the incidental finding that this pore-forming protein itself greatly weakens the membrane in doses lower than required for effective permeabilization.