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Local calcium release in mammalian skeletal muscle

Natalia Shirokova, Jesús García * and Eduardo Ríos

1. Fluo-3 fluorescence associated with Ca²⁺ release was recorded with confocal microscopy in single muscle fibres mechanically dissected from fast twitch muscle of rats or frogs, voltage clamped in a two Vaseline-gap chamber.

2. Interventions that elicited Ca²⁺ sparks in frog skeletal muscle (low voltage depolarizations, application of caffeine) generated in rat fibres images consistent with substantial release from triadic regions, but devoid of resolvable discrete events. Ca²⁺ sparks were never observed in adult rat fibres. In contrast, sparks of standard morphology were abundant in myotubes from embryonic mice.

3. Depolarization-induced gradients of fluorescence between triadic and surrounding regions (which are proportional to Ca²⁺ release flux) peaked at about 20 ms and then decayed to a steady level. Gradients were greater in frog fibres than in rat fibres. The ratio of peak over steady gradient (R) was steeply voltage dependent in frogs, reaching a maximum of 4·8 at -50 mV (n = 7). In rats, R had an essentially voltage-independent value of 2·3 (n = 5).

4. Ca²⁺-induced Ca²⁺ release, resulting in concerted opening of several release channels, is thought to underlie Ca²⁺ sparks and the peak phase of release in frog skeletal muscle. A diffuse 'small event' release, similar to that observed in these rats, is also present in frogs and believed to be directly activated by voltage. The present results suggest that in these rat fibres there is little contribution by CICR to Ca²⁺ release triggered by depolarization, and a lack of concerted channel opening.

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