Isolated cardiac muscle preparations suffer from damaged-end compliance that allows for substantial shortening of central sarcomeres during contractions in which the overall length of muscle is kept constant. The impact of uncontrolled sarcomere shortening during a twitch on the intracellular calcium transient in myocardium is unknown. Accordingly, in the present study we developed an iterative laser-diffraction feedback system that allowed for the accurate control of central-segment sarcomere length and simultaneous measurement of iontophoretically injected fura 2 fluorescence in isolated cardiac trabeculae. We compared fura 2 fluorescence signals recorded during regular twitches with twitches in which central sarcomere length (SL) was held constant by feedback control ("SL clamp" twitches). We found that uncontrolled sarcomere shortening was associated with a significant (P = 0.005) increase in the peak of the calcium transient and that the amount of this increase was directly correlated to the extent of central-segment sarcomere shortening (r2 = 0.92; P < 0.01). The time course of the calcium transient, however, was unaffected by the mode of contraction (P = 0.64). These findings have important implications for the interpretation of studies of myocardial calcium handling in which uncontrolled sarcomere shortening takes place during the twitch.

The effect of sudden local fluctuations of the free sarcoplasmic [Ca++]i in cardiac cells on calcium release and calcium uptake by the sarcoplasmic reticulum (SR) was calculated with the aid of a simplified model of SR calcium handling. The model was used to evaluate whether propagation of calcium transients and the range of propagation velocities observed experimentally (0.05-15 mm s -1) could be predicted. Calcium fluctuations propagate by virtue of focal calcium release from the SR, diffusion through the cytosol (which is modulated by binding to troponin and calmodulin and sequestration by the SR), and subsequendy induce calcium release from adjacent release sites of the SR. The minimal and maximal velocities derived from the simulation were 0.09 and 15 mm s -1 respectively. The method of solution involved writing the diffusion equation as a difference equation in the spatial coordinates. Thus, coupled ordinary differential equations in time with banded coefficients were generated. The coupled equations were solved using Gear's sixth order predictor-corrector algorithm for stiff equations with reflective boundaries. The most important determinants of the velocity of propagation of the calcium waves were the diastolic [Ca++]i, the rate of rise of the release, and the amount of calcium released from the SR. The results are consistent with the assumptions that calcium loading causes an increase in intraceUular calcium and calcium in the SR, and an increase in the amount and rate of calcium released. These two effects combine to increase the propagation velocity at higher levels of calcium loading.

We studied contractile function in cardiac trabeculae isolated from the right ventricles (RV) of rats with experimental heart failure (HF) induced by left ventricular (LV) myocardial infarction (24 wk post-MI; n = 6) and from sham-operated rats (n = 7). Sarcomere length (SL) was measured by laser diffraction techniques, and force (F) was measured by silicon strain gauge. SL was kept constant at all times by computer feedback control. HF was associated with marked LV dilation and pulmonary congestion. In intact, RV twitching trabeculae, HF was associated with a depression of the F-SL relation at extracellular Ca2+ concentration ([Ca2+]o) = 1.5 mM and a depression of the F-[Ca2+]o relation at SL = 2.0 microns. HF was also associated with a significant depression of the F-intracellular [Ca2+] relation at SL = 2.0 microns measured after chemical permeabilization of these RV trabeculae (skinned fibers). Our results suggest that reduced force development in this model of HF is due, in part, to depressed function of the contractile filaments.