Basal and acidic fibroblast growth factor-induced atrial natriuretic peptide gene expression and secretion is inhibited by staurosporine.
We examined the mechanisms involved in the activation of atrial natriuretic peptide (ANP) gene expression and secretion in response to acidic fibroblast growth factor (aFGF) by studying the effects of staurosporine, a protein kinase C inhibitor, and 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C, on basal and AFGF-induced ANP messenger RNA (mRNA) and immunoreactive ANP (IR-ANP) levels in cultured neonatal rat cardiac myocytes. Acidic FGF caused a dose- and time-dependent increase in IR-ANP and immunoreactive N-terminal fragment of proANP (IR-NT-proANP) release into the culture medium from ventricular but not from atrial myocytes. In ventricular cells, 50 ng/ml aFGF for 24 or 48 h resulted in a 70% or 181% increase, respectively, in the accumulation of IR-ANP into the culture medium. Acidic FGF also stimulated ANP gene expression significantly; after 48 h of incubation, the ANP mRNA levels of aFGF-treated ventricular myocytes were 205% (P < 0.001) higher than those of control cells. Staurosporine alone at concentration of 10 nM significantly decreased the basal IR-ANP and IR-NT-proANP secretion, and inhibited the aFGF-induced increase in ANP mRNA and IR-ANP levels in ventricular myocytes. TPA (100 nM) alone significantly stimulated ANP gene expression and secretion but these effects were not augmented by combining aFGF with TPA. High performance liquid chromatographical analysis showed that atrial and ventricular myocytes maintained in serum-free medium were capable of secreting processed, ANP99-126 sized material, and that aFGF did not alter the processing of ANP in ventricular cultures. These results demonstrate that aFGF is a potent stimulator of ANP gene expression and secretion in cultured neonatal rat ventricular but not in atrial cells. The observations that (a) staurosporine completely abolished the effects of aFGF on ANP gene expression and release and (b) ANP secretory and gene expression inducing effects of phorbol ester were not augmented by aFGF, suggest an important role of protein kinase C in mediating aFGF-induced ANP gene expression and secretion.
Tokola
H
H
Department of Pharmacology and Toxicology, University of Oulu, Finland.
Salo
K
K
Vuolteenaho
O
O
Ruskoaho
H
H
D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D016220:Fibroblast Growth Factor 1 / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects*; D006352:Heart Ventricles / Q000187:drug effects / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000470:Alkaloids; D003470:Culture Media; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D009320:Atrial Natriuretic Factor; D011493:Protein Kinase C; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate