Phorbol Myristate Acetate-induced Hypertrophy of Neonatal Rat Cardiac Myocytes is Associated with Decreased Sarcoplasmic Reticulum CaATPase (SERCA2) Gene Expression and Calcium Reuptake |
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Authors: | Ron Hartong Francisco J. Villarreal Frank Giordano Randa Hilal-Dandan Patrick M. McDonough Wolfgang H. Dillmann |
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Affiliation: | Department of Medicine, University of California San Diego, San Diego, CA, USA;Department of Biology, San Diego State University, San Diego, CA, USA |
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Abstract: | The decreased expression of the sarcoplasmic reticulum Ca2+-ATPase associated with cardiac hypertrophy was investigated in cultured neonatal rat cardiac myocytes. Northern blot analysis indicated a significant 55–60% decrease in Ca2+-ATPase mRNA levels and after 12 and 24 h of treatment with the phorbol ester phorbol myristate acetate (PMA). Myocytes treated with the phorbol ester for 80 h showed a significant 34% decrease (relative to vehicle-treated control cells) in the levels of Ca2+-ATPase protein, and a significant 38% increase in the levels ofα-sarcomeric actin, as assessed by Western blot analysis using specific antibodies. Immunocytochemistry of myocytes treated for 72 h with the phorbol ester revealed a hypertrophied cell morphology, and showed a marked decrease in Ca2+-ATPase staining intensity. Contractile calcium transients were evaluated through the use of indo-1. It was found that thet1/2for the decline of calcium transient was significantly prolonged by PMA treatment (0.51±0.15) when compared to controls (0.38±0.17,P<0.001). Treatment of myocytes with endothelin-1 also led to a 35% decrease in sarcoplasmic reticulum Ca2+-ATPase mRNA levels. It is concluded that phorbol ester treatment of neonatal rat cardiac myocytes induces similar changes in Ca2+-ATPase gene expression as observedin vivoin the hypertrophied and failing heart. The observed prolongation int1/2for [Ca2+]idecline might be due to the observed depressed levels for sarcoplasmic reticulum Ca2+-ATPase in PMA treated cells. |
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Keywords: | Ca2+-ATPase Endothelin-1 Sarcoplasmic reticulum Cardiac myocytes Phorbol esters Cardiac hypertrophy Gene expression Calcium reuptake |
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