Alterations in intracellular calcium and tension of activated ferret papillary muscle in response to step length changes

1. To study the effects of mechanical constraints on the calcium (Ca2+) affinity of cardiac troponin C, we analysed the tension and aequorin light (AL, intracellular Ca2+) transients in response to a step length change in aequorin-injected ferret right ventricular papillary muscles. The muscle preparations were continuously activated with ouabain (10(-4) M) (ouabain contracture) or with high frequency stimuli in the presence of ryanodine (5 microM) (tetanic contraction). 2. The tension transient in response to either the release or stretch was oscillatory: tension decreased rapidly during the release and then increased, after which it lapsed into a new steady level in a series of damped oscillations. The opposite was true for the stretch. The oscillatory responses were conspicuous and less damped in ouabain-activated preparations (oscillation frequency of 2.2-2.3 Hz at 22 degrees and 4.5-4.6 Hz at 30 degrees C) and much more damped in ryanodine-treated preparations. 3. The transient AL response was also oscillatory, the time course of which corresponded to that of the transient tension response. Regardless of the difference in the time course of the transients in two different preparations and at two different temperatures, the increase in AL corresponded to the decrease in tension, likewise the decrease in AL to the increase in tension. 4. The mean level of AL after release was lower than the control level present just prior to the release in ouabain-activated preparations, but the AL after release finally returned to the nearly control level in ryanodine-treated preparations. 5. When the ryanodine-treated muscle was further treated with 2,3-butanedione monoxime (BDM) (20 mM), the tetanic tension decreased remarkably without affecting the AL signal. The tension transient of this preparation was quite similar to that of the resting muscle, which changed in a nearly stepwise fashion; AL was hardly affected by step length changes, as in the resting muscle, in spite of the higher AL level. 6. These results suggest that the Ca2+ affinity of cardiac troponin C is increased with an increase in tension (i.e. the cross-bridge attachment) and decreased with a decrease in tension i.e. the cross-bridge detachment), and that the mean [Ca2+]i is lowered by release, at least in a Ca(2+)-overloaded condition, mainly through the sarcoplasmic reticulum.