Changes in phosphoinositide turnover in isolated guinea pig hearts stimulated with isoproterenol.
The incorporation of 32Pi into phospholamban, troponin I, phosphatidylinositols, and inositol trisphosphates was studied in Langendorff-perfused guinea pig hearts stimulated with isoproterenol. Hearts were perfused with Krebs-Henseleit buffer containing [32P]Pk and freeze-clamped at different times during the positive inotropic response. Exposure of the hearts to 0.1 microM isoproterenol for up to 1 minute was associated with significant (up to threefold) increases in phospholamban and troponin I phosphorylation, but there was no significant increase in 32P incorporation into phospholipids. However, longer exposure (2 minutes or more) to isoproterenol was associated with increases in the degree of 32P labeling of phosphatidylinositols and phosphatidic acid. Examination of 32P labeling of inositol trisphosphates in the same hearts revealed that the radioactivity associated with these compounds decreased with time. The decreases were significant at times of exposure of 2 minutes or longer to beta-adrenergic stimulation. The tissue levels of the inositol 1,4,5-trisphosphate isoform were also measured in hearts perfused with isoproterenol for 3 minutes, and they were found to be significantly lower compared with values obtained in control hearts. The effects of isoproterenol on 32P incorporation into phospholipids and proteins were observed in the presence of prazosin, and they were completely abolished by the beta-receptor blocker propranolol. Examination of the phosphoinositide-specific phospholipase C activity in the perfused hearts revealed that isoproterenol stimulation was associated with a decrease in the membrane-associated enzymatic activity at physiological calcium concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)
Edes
I
I
Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Ohio.
Solaro
R J
RJ
Kranias
E G
EG
D000818:Animals; D001345:Autoradiography; D002135:Calcium-Binding Proteins / Q000096:biosynthesis; D002855:Chromatography, Thin Layer; D006168:Guinea Pigs; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology*; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D009994:Osmolar Concentration; D010716:Phosphatidylinositols / Q000378:metabolism*; D010743:Phospholipids / Q000096:biosynthesis; D010758:Phosphorus / Q000378:metabolism; D013268:Stimulation, Chemical; D013997:Time Factors; D014336:Troponin / Q000096:biosynthesis; D019210:Troponin I; D000099211:Phospholamban
D002135:Calcium-Binding Proteins; D015544:Inositol 1,4,5-Trisphosphate; D007545:Isoproterenol; D010716:Phosphatidylinositols; D010743:Phospholipids; D010758:Phosphorus; D014336:Troponin; D019210:Troponin I; D000099211:Phospholamban