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Volume 272, Number 10, Issue of March 7, 1997 pp. 6685-6692
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Opioid Peptide Gene Expression in the Primary Hereditary Cardiomyopathy of the Syrian Hamster
I. REGULATION OF PRODYNORPHIN GENE EXPRESSION BY NUCLEAR PROTEIN KINASE C

(Received for publication, July 5, 1996, and in revised form, November 18, 1996)

Carlo Ventura ^§ , Gianfranco Pintus ^§ , Maria Giovanna Fiori , Federico Bennardini , Giangavino Pinna and Leonardo Gaspa ^§

From the  Institute of Biological Chemistry "A. Bonsignore," School of Medicine, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy, the ^§ National Laboratory of the National Institute of Biostructures and Biosystems, Osilo, Italy, and the  Institute of Biological Chemistry, School of Pharmacy, University of Sassari, Via Muroni 23/A, 07100 Sassari, Italy

Prodynorphin gene expression was investigated in adult ventricular myocytes isolated from normal (F1B) or cardiomyopathic (BIO 14.6) hamsters. Prodynorphin mRNA levels were higher in cardiomyopathic than in control myocytes and were stimulated by treatment of control cells with the protein kinase C (PKC) activator 1,2-dioctanoyl-sn-glycerol. Both chelerythrine and calphostin C, two PKC inhibitors, abolished the stimulatory effect of the diglyceride and significantly reduced prodynorphin gene expression in cardiomyopathic myocytes. Nuclear run-off experiments indicated that the prodynorphin gene was regulated at the transcriptional level and that treatment of nuclei isolated from control cells with 1,2-dioctanoyl-sn-glycerol increased prodynorphin gene transcription, whereas chelerythrine or calphostin C abolished this transcriptional effect. Direct exposure of nuclei isolated from cardiomyopathic myocytes to these inhibitors markedly down-regulated the rate of gene transcription. The expression of PKC-, -, and -, as well as PKC activity, were increased in nuclei of cardiomyopathic myocytes compared with nuclei from control cells. The levels of both intracellular and secreted dynorphin B, a biologically active product of the gene, were higher in cardiomyopathic than in control cells and were stimulated or inhibited by cell treatment with 1,2-dioctanoyl-sn-glycerol or PKC inhibitors, respectively.

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