Gq/11-coupled receptors and protein synthesis in rat cardiomyocytes: role of Gi-proteins and protein kinase C-isozymes.

Abstract

Our recent findings indicate that, in rat neonatal ventricular cardiomyocytes, endothelin-1 (ET-1) induces increases in the rate of protein synthesis in a partly pertussis toxin (PTX)-sensitive manner, and that angiotensin II-evoked increases in the rate of protein synthesis are brought about via local secretion of ET-1. The aim of this study was to find out: (1) whether noradrenaline (NA) and the thromboxane A2 (TXA2)-mimetic U 46619-induced increases in the rate of protein synthesis may be also partly PTX-sensitive and/or mediated by ET-1, and (2) whether the growth-promoting effects of NA and U 46619 as well as ET-1 might involve activation of the same set of protein kinase C (PKC) isozymes. For this purpose we first studied the effects of NA and U 46619 on inositol phosphate (IP)-formation (assessed as accumulation of total [3H]IPs in myo-[3H]inositol prelabelled cells) and on the rate of protein synthesis (assessed as [3H]phenylalanine incorporation) (1) in the presence and absence of the ET(A)-receptor antagonist BQ-123, and (2) in nontreated and PTX-pretreated cells. Second, we assessed the effects of the PKC-inhibitors bisindolylmaleimide I and Gö 6976 and of phorbol-12-myristate-13-acetate (PMA; 1 microM overnight)-pretreatment on U 46619-, NA- and ET-1-induced increases in the rate of protein synthesis. NA (0.01-10 microM) concentration-dependently increased IP-formation (maximum increase: 115-/+23% above basal, n=4) and [3H]phenylalanine incorporation (maximum increase: 40+/-3% above basal, n=20). Both responses were antagonized by the alpha1-adrenoceptor antagonist prazosin (1 microM), but were not significantly affected by BQ-123 (1 microM). U 46619 (0.01-100 microM) concentration-dependently increased IP-formation (maximum increase: 89+/-12% above basal, n=8) and [3H]phenylalanine incorporation (maximum increase: 33+/-4% above basal, n=16). Both responses were slightly but significantly antagonized by the TP-receptor antagonist SQ 29548 (1 microM), but were not affected by BQ-123 (1 microM). Pretreatment of the cardiomyocytes with 250 ng ml(-1) PTX overnight did not significantly affect NA- and U 46619-evoked increases in IP-formation and [3H]phenylalanine incorporation. The PKC-inhibitor bisindolylmaleimide I (5 microM) as well as pretreatment of the cells with PMA (1 microM) significantly reduced the effects of NA, U 46619 and ET- I on the rate of protein synthesis; in contrast, the PKC-inhibitor Gö 6976 (5 microM) was without any effects. We conclude that, in rat neonatal ventricular cardiomyocytes, stimulation of Gq/11-coupled receptors increases the rate of protein synthesis; this involves activation of the same PKC-isozymes (very likely PKC-delta and/or -epsilon). NA and U 46619 cause their growth-promoting effects in a PTX-insensitive manner; ET-1 is not involved in their effects.