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  • Title: Contractile activity modulates atrial natriuretic factor gene expression in neonatal rat ventricular myocytes.
    Author: Eble DM, Cadre BM, Qi M, Bers DM, Samarel AM.
    Journal: J Mol Cell Cardiol; 1998 Jan; 30(1):55-60. PubMed ID: 9500864.
    Abstract:
    [Ca2+]i transients, and the activation of Ca(2+)-sensitive kinases have been considered potential signaling mechanisms regulating ANF gene expression in cultured neonatal rat ventricular myocytes (NRVM). However, it is unclear whether [Ca2+]i is directly involved, or is indirectly involved by generating additional mechanical signals via contractile activity. Primary cultures of spontaneously contracting NRVM (CON), and NRVM treated for 48 h with verapamil (V, 10 microM), KCl (50 mM), or 2,3-butanedione monoxime (BDM, 7.5 mM) were used to delineate the affects of contractile activity v [Ca2+]i. Verapamil, a calcium, channel blocker, inhibits contraction and decreases [Ca2+]i. High [K+]o causes membrane depolarization, loss of contraction, and elevates [Ca2+]i; whereas BDM strongly inhibits contractile activity but only modestly reduces [Ca2+]i transients. ANF production, as assessed by radioimmunoassay, was significantly reduced upon contractile arrest independently of [Ca2+]i levels. Northern blotting analysis demonstrated that contractile arrest also reduced ANF mRNA levels. Transient transfection of a 3003 bp ANF promoter-luciferase expression plasmid in CON, V, KCl, and BDM-treated NRVM demonstrated marked down-regulation of ANF promoter activity in all of the contractile arrested myocytes. These results indicate that the activation of Ca(2+)-sensitive processes alone are insufficient to maintain high levels of ANF gene expression and peptide production in NRVM.
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