Mechanism of the negative force-frequency relationship in physiologically intact rat ventricular myocardium--studies by intracellular Ca2+ monitor with indo-1 and by 31P-nuclear magnetic resonance spectroscopy.

We studied the subcellular mechanisms of the negative force-frequency relationship in rat myocardium by measuring 1) intracellular Ca2+ transients by indo-1 fluorometry and 2) intracellular pH (pHi) and phosphate compounds with 31P-nuclear magnetic resonance (NMR). The data were compared with those from guinea pig hearts, which show a positive force-frequency relationship. By increasing the pacing rate from 3 Hz to 5 Hz, the peak positive first derivative of left ventricular pressure (LVdP/dt) in rat heart decreased by 10 +/- 1% (n = 6). In contrast to this negative inotropic response, simultaneously measured peak Ca2+ transients increased by 6 +/- 1%. Guinea pig heart (n = 6) showed an increase in peak positive LVdP/dt (33 +/- 1%) which was associated with an increase in peak Ca2+ transients (8 +/- 1%). Under equivalent experimental conditions in an NMR spectrometer, this increase in the pacing rate did not affect intracellular levels of phosphate compounds in either rat (n = 6) or guinea pig heart (n = 6). In contrast, pHi showed a decrease of 0.031 +/- 0.006 pH units in rat heart, while no changes were observed in guinea pig heart. These results suggest that in physiological rat myocardium, pHi is susceptible to changes in the stimulus frequency and may affect the Ca(2+)-responsiveness of contractile proteins, which results in the negative force-frequency relationship.