Identification of the molecular basis for phosphorylase hypersensitivity in cultured diabetic cardiomyocytes.
The focus of this study was to identify the molecular basis for the hypersensitive response of glycogen phosphorylase activation to epinephrine stimulation in alloxan diabetic-derived cardiomyocytes. Cyclic AMP levels were found not to be significantly different between normal and diabetic-derived cells while cGMP concentrations were found consistently to be significantly lower in diabetic-derived cells than in normal cells. Treatment with cyclic GMP analogues did not affect phosphorylase activation by epinephrine in normal cardiomyocytes whereas, IBMX, a nonselective phosphodiesterase inhibitor, had a significant effect on basal and agonist-stimulated phosphorylase activity in both normal and diabetic-derived cardiomyocytes. Differences in the time course for the rate of decay of phosphorylase a from agonist-stimulated to basal levels were observed between normal and diabetic cells. After 3 h in primary culture, phosphorylase a activity returned to basal levels more quickly in normal than in diabetic-derived cells while after 24 h in culture, the time for phosphorylase a decay was not significantly different between normal and diabetic myocytes and was longer than the 3 h response. After 3 h response. After 3 h in primary culture, no significant difference in phosphorylase kinase activity was observed between normal and diabetic-derived cells exposed to epinephrine whereas, after 24 h in culture, phosphorylase kinase activity was significantly decreased in diabetic cells under basal and agonist-stimulation conditions.(ABSTRACT TRUNCATED AT 250 WORDS)
Buczek-Thomas
J A
JA
Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School, Worcester 01655, USA.
Miller
T B
TB
D000818:Animals; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism* / Q000473:pathology; D004342:Drug Hypersensitivity / Q000201:enzymology* / Q000473:pathology; D004789:Enzyme Activation; D004837:Epinephrine / Q000494:pharmacology*; D006321:Heart / Q000187:drug effects; D008297:Male; D009206:Myocardium / Q000201:enzymology* / Q000473:pathology; D006005:Phosphorylases / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013268:Stimulation, Chemical
D006005:Phosphorylases; D004837:Epinephrine