Birefringence changes associated with isometric contraction and rapid shortening steps in frog skeletal muscle fibres

1. Muscle birefringence, the difference between the refractive indices of light polarized parallel and perpendicular to the muscle fibre axis, was measured at 3 degrees C in intact single fibres isolated from frog muscle. Resting birefringence was 2.20 +/- 0.02 x 10(-3) (mean +/- S.E.M., n = 44) at sarcomere length 2.4-2.7 microns and 2.35 +/- 0.03 x 10(-3) (n = 19) at 3.5-3.8 microns. 2. Birefringence decreased during isometric twitch or tetanic contractions. The peak change in a twitch at sarcomere length 2.6 microns, determined by two independent methods, was 0.150 +/- 0.017 x 10(-3) (mean +/- S.E.M., n = 6). The corresponding value after 0.4 s of tetanic stimulation was 0.167 +/- 0.012 x 10(-3) (n = 6). 3. The birefringence change had a shorter latency than tension and reached its half-maximum value earlier than tension. The difference in time to half-maximum in tetani was 11.5 +/- 1.3 ms (mean +/- S.E.M., n = 6) at 3 degrees C. After stimulation birefringence recovered to its pre-stimulus baseline more slowly than tension. 4. The birefringence decrease after 0.4 s of tetanic stimulation was linearly related to the expected degree of overlap between actin and myosin filaments in the sarcomere length range 2.6-3.6 microns. The amplitude of the birefringence decrease at full filament overlap (sarcomere length 2.2 microns) was estimated to be 0.235 +/- 0.015 x 10(-3). 5. Birefringence changes associated with shortening steps of 0.9% fibre length at sarcomere length 2.6 microns exhibited four phases corresponding to those of the tension transient. There was no consistent birefringence change during the length step itself. During the rapid tension recovery birefringence increased by 0.014 +/- 0.001 x 10(-3) (n = 3), measured from the end of the length step to 2 ms later. Birefringence continued to increase as tension recovery slowed, reaching a peak about 10 ms after the step, then recovered with a rate similar to that of the final tension recovery. 6. These birefringence changes are likely to be caused by axial rotation of the head domain of the myosin cross-bridge. During isometric contraction heads bind to actin with their long axes more perpendicular to the fibre axis than in resting muscle, although there is likely to be a wide range of head orientations during contraction.(ABSTRACT TRUNCATED AT 400 WORDS)