The effect of changing free Ca²⁺ on light diffraction intensity and correlation with tension development in skinned fibers of frog skeletal muscle

Abstract

The relationship between the diffraction intensity change of the first order line and tension development was examined in mechanically skinned single fibers from the dorsal head of the semitendinosus of frogs. Passive stretch of the fibers resulted in an increase in intensity over the range of sarcomere lengths from 2.5 to 3.6 μm, indicating that the intensity is a function of sarcomere length. Activation of skinned fibers caused a decrease in the intensity, at all sarcomere lengths, where the thick and thin filaments overlapped. The magnitude of the intensity decrease and that of the tension development depended on the Ca²⁺ concentration in the medium. The drop of intensity-pCa and the tension-pCa curves showed a similarly steep S-shape within a range of 0.5 pCa unit, although the intensity-pCa curve shifted to the left; the pCa for 50% decrease in light signal was 6.48 and that for 50% tension development was 6.40. Caffeine (25 mM) added to the medium produced a decrease in the intensity of skinned fibers with the simultaneous development of tension, thereby indicating that caffeine induces a release of Ca²⁺ from the sarcoplasmic reticulum and disorder in the filaments ensues. Changes in diffraction intensity with electrical stimulation to the intact single fiber were similar, although a more striking summation was observed in the optical response, as compared to the tension development. These results suggest that tension development upon stimulation can be monitored by assessing the magnitude of diffraction intensity decrease in the first order line, except for some shift in the short fiber.