Diastolic, systolic and sarcoplasmic reticulum [Ca2+] during inotropic interventions in isolated rat myocytes

1. The fluorescent indicator Fura-2 has been used to monitor intracellular [Ca2+] (Ca2+i) in myocytes isolated from the ventricles of rat hearts. 2. The relationships between diastolic Ca2+i, systolic Ca2+i and the Ca2+ content of the sarcoplasmic reticulum (SR; assayed using caffeine) have been studied during changes of stimulation rate and bathing [Ca2+] (Ca2+o). 3. When stimulation rate was increased, there were increases in diastolic Ca2+i, systolic Ca2+i and the Ca2+ content of the SR. 4. The SR inhibitor ryanodine (1 mumol l-1) decreased the size of the Ca2+i transient, and abolished the increase of Ca2+i produced by caffeine (10 mmol l-1). In the presence of ryanodine, increasing stimulation rate increased diastolic Ca2+i but not systolic Ca2+i. 5. Increasing Ca2+o led to increases of diastolic Ca2+i, systolic Ca2+i and SR Ca2+ content similar to those observed during changes in stimulation rate. 6. Ryanodine altered the relationship between systolic and diastolic Ca2+i during changes of Ca2+o. 7. These results are consistent with a change of diastolic Ca2+i leading to an increase in the Ca2+ content of the SR, and hence an increase in the size of the Ca2+i transient during changes in stimulation rate and Ca2+o.