Vol. 58, Issue 6, 1546-1553, December 2000

Low Catecholamine Concentrations Protect Adult Rat Ventricular Myocytes against Apoptosis through cAMP-Dependent Extracellular Signal-Regulated Kinase Activation

Morgana Henaff, Stéphane N. Hatem, and Jean-Jacques Mercadier

Institut National de la Santé et de la Recherche Médicale Unité 460, Faculté de Médecine Xavier Bichat, Paris, France

Catecholamines have complex effects on cardiac myocyte growth and survival, including the triggering of apoptosis at high concentration. Here, we examined whether at a lower concentration, catecholamine protected adult rat ventricular myocytes from apoptosis in vitro. Myocytes were exposed to staurosporine (ST, 10 µM) for 18 h, with or without epinephrine (0.1 or 10 µM) or fetal calf serum (10%). Apoptosis was assessed after 48 h of culture in terms of DNA fragmentation (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method, DNA gel electrophoresis). Epinephrine (0.1 µM) and serum reduced ST-induced myocyte apoptosis by ~50% (n = 12 cultures, P < .001), whereas epinephrine and serum alone did not influence the low apoptotic rate in control cultures. In contrast, 10 µM epinephrine induced marked apoptosis in ST-free conditions. The protective effects of 0.1 µM epinephrine and serum were blunted by the tyrosine kinase inhibitor genistein (n = 12 cultures, P < .001). Extracellular signal-regulated kinase (ERK) activity was stimulated by 0.1 µM epinephrine but not by 10 µM epinephrine. Furthermore, the protective effect of epinephrine was mimicked by isoproterenol (1 µM) and forskolin (1 µM) but not by phenylephrine (10 µM) and was blunted by propranolol (10 µM) but not by prazozin (10 µM). Finally, isoproterenol and forskolin activated ERK, an effect that was blunted by propranolol. In conclusion, low epinephrine concentrations attenuate ST-induced apoptosis of adult cardiac myocytes in vitro, an effect mediated by coupling between the cAMP pathway and ERK activation. This suggests that a minimal adrenergic tone is essential for myocyte survival in conditions of unusual stress.