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Identification of the molecular basis for phosphorylase hypersensitivity in cultured diabetic cardiomyocytes

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Abstract

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 phosphorylasea from agonist-stimulated to basal levels were observed between normal and diabetic cells. After 3 h in primary culture, phosphorylasea activity returned to basal levels more quickly in normal than in diabetic-derived cells while after 24 h in culture, the time for phosphorylasea decay was not significantly different between normal and diabetic myocytes and was longer than the 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-stimulated conditions. These data collectively suggest that the hypersensitive response of glycogen phosphorylase to epinephrine stimulation in diabetic-derived cardiomyocytes is not due to a defect present at the level of phosphorylase kinase but may, in part, result from an alteration in cardiac phosphodiesterase activity resulting from diminished intracellular cyclic GMP concentrations.

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  1. Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School, 55 Lake Avenue North, 01655, Worcester, Massachusetts, USA

    Jo Ann Buczek-Thomas & Thomas B. Miller Jr.

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  1. Jo Ann Buczek-Thomas
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  2. Thomas B. Miller Jr.
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Buczek-Thomas, J.A., Miller, T.B. Identification of the molecular basis for phosphorylase hypersensitivity in cultured diabetic cardiomyocytes. Mol Cell Biochem 145, 131–139 (1995). https://doi.org/10.1007/BF00935485

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  • DOI: https://doi.org/10.1007/BF00935485

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