Background: An increase in cytosolic protein phosphatases (PPs) de-phosphorylates phospholamban, decreasing the Ca(2+) uptake of the sarcoplasmic reticulum (SR). The effects of PP inhibitors on cellular Ca(2+) handling were investigated.
Methods and results: Twitch Ca(2+) transients (CaTs) and cell shortening were measured in intact rat cardiac myocytes, and caffeine-induced Ca(2+) transients (CaffCaTs) and Ca(2+) sparks were studied in saponin-permeabilized cells. Calyculin A augmented isoproterenol-induced increases in CaTs and cell shortening without altering the diastolic [Ca(2+)](i) and twitch [Ca(2+)](i) decay. The protein kinase A catalytic subunit (PKA(cat)) increased the peak of CaffCaTs between 5 and 50 U/ml, and the addition of inhibitor-1 (I-1) augmented the increase. PKA(cat) increased Ca(2+) spark frequency and the addition of I-1 increased it further. PKA(cat) at 50 U/ml amplified the peak and prolonged the duration of Ca(2+) sparks, whereas the addition of I-1 did not alter them. An abrupt inhibition of SR Ca(2+) uptake following exposure to PKA(cat) caused a gradual decrease in Ca(2+) spark frequency, but the addition of I-1 did not accelerate the decline of Ca(2+) spark frequency or CaffCaTs.
Conclusions: Inhibition of PPs augmented the inotropic effect of isoproterenol. Specific inhibition of PP1 could stimulate the Ca(2+) uptake of the SR with less significant effects on the Ca(2+) release.