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Circulation, Vol 85, 1835-1841, Copyright © 1992 by American Heart Association

ARTICLES

Gene expression and atrial natriuretic factor processing and secretion in cultured AT-1 cardiac myocytes

NA Lanson Jr, CC Glembotski, ME Steinhelper, LJ Field and WC Claycomb
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, New Orleans 70112.

BACKGROUND. Studies were carried out to characterize several biochemical features of cultured AT-1 cells. METHODS AND RESULTS. These cells were obtained from a transplantable atrial cardiomyocyte tumor lineage. Reverse transcriptase-polymerase chain reaction-based analyses demonstrated that the pattern of gene expression of cultured AT-1 cells was similar to that of adult atrial myocytes. AT-1 cells expressed atrial natriuretic factor (ANF), alpha-cardiac myosin heavy chain, alpha-cardiac actin, and connexin43. Radioimmunoassays verified that the cells synthesized, stored, and secreted ANF. Through size- exclusion, reversed-phase, and carboxymethyl-ion-exchange high- performance liquid chromatography, it was shown that cultured AT-1 cells stored ANF as pro-ANF (ANF-[1-126]), which was cosecretionally processed quantitatively to ANF-(1-98) and the bioactive 28-amino-acid ANF-(99-126). In addition, cultured AT-1 cells secreted ANF at almost a sixfold greater rate in response to endothelin-1, a potent secretagogue of ANF. KCl, metenkephalinamide, isoproterenol, phenylephrine, and 12-O- tetradecanoyl-phorbol-13-acetate also stimulated ANF release. CONCLUSIONS. These studies, in combination with previous findings, demonstrated that cultured AT-1 cells, while maintaining the ability to proliferate, have retained functional, biochemical, and ultrastructural features that are characteristic of adult atrial myocytes.

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