Effect of growth hormone and inhibitors of protein kinase A on IGF-I, oxytocin and progesterone release by cultured bovine granulosa cells

In our experiments we studied the action of GH on the release of nonapeptide, steroid hormones and growth factor, in bovine ovarian granulosa cells, as well as the role of cAMP-stimulated protein kinase A (PKA) in the mediation of these GH effects. For this purpose, the effects of exogenous bGH (0.001-10 mg/ml), PKA blockers KT5720 (100 ng/ml) and Rp-cAMPS (1 mmol), alone and in combination, on IGF-I, oxytocin and progesterone secretion were investigated. It was found that GH addition to culture medium strongly (p<0.05) stimulated IGF-I (at a concentration of 0.01-0.1 mgGH/ml medium), oxytocin (0.01-10 mgGH/ml) and progesterone (0.01-1 mgGH/ml medium) secretion into the culture medium. PKA blockers KT5720 and Rp-cAMPS given alone did not affect release of these substances. Rp-cAMPS partially prevented GH effect on IGF-I release, but enhanced GH action on progesterone output. KT5720 did not modify action of GH on oxytocin release. These observations confirm the involvement of GH in the control of IGF-I, oxytocin and progesterone release by bovine ovarian granulosa cells. Effects of PKA blocker on several GH-induced effects suggest that GH effects on IGF-I and progesterone, but not on oxytocin release may be partially mediated by the cAMP/PKA-dependent intracellular mechanisms.