Abstract

Cultured hind limb skeletal muscle cells from newborn rats were used to study the effect of caffeine and tetracaine upon intracellular Ca2+ release under voltage or current clamp conditions. Free [Ca2+]i was measured using the fluorescent calcium-sensitive dye Fluo-3. A field containing one or several myotubes was observed with a video camera and image analysis of fluorescence changes was performed. Addition of 100-500 microM tetracaine to the external saline elicited strong fluorescence responses in non-clamped cells, but significantly lower responses in cells clamped at -90 mV. At the same time, tetracaine inhibited voltage induced calcium release. Voltage and tetracaine modulation over the action of caffeine (500 microM) was also observed. Pretreatment of cells with 10 microM nifedipine abolished the caffeine induced fluorescence response in non-clamped cells. These findings suggest that, in cultured muscle cells, calcium release through the caffeine and tetracaine sensitive pathways is controlled by both membrane potential and the dihydropyridine receptor.