Growth hormone and insulin-like growth factor-I action on progesterone secretion by porcine corpora lutea isolated at various periods of the luteal phase

This study was conducted to investigate the interactions between growth hormone (GH) and insulin-like growth factor-I (IGF-I) on progesterone (P4) secretion by porcine luteal cells cultured in vitro. Cells isolated from corpora lutea (CL) collected at three different periods of the luteal phase (CL1 - early luteal phase; CL2 - middle luteal phase and CL3 - late luteal phase) were incubated with different doses of GH (10, 100 or 200 ng/ml). After 48 h cultures were terminated and the media were frozen until further P4 concentration analysis. GH (100 ng/ml) increased P4 secretion by CL1 and CL2 and had no effect on CL3. In separate studies these cells were treated for 48 h with IGF-I alone or with GH combined with IGF-I. IGF-I alone increased basal P4 secretion only by cells collected from CL1 while concurrent treatment with GH had no effect on P4 secretion by any type of CL. To investigate the possible mechanism of GH and IGF-I mediated induction of P4 secretion, an inhibitory study was conducted. In this experiment, luteal cells collected from CL1 were cultured in the absence or presence of cycloheximide (an inhibitor of protein synthesis) or actinomycin D (an inhibitor of DNA transcription). Cycloheximide or actinomycin D completely blocked the stimulatory effect of both GH and IGF-I on P4 production but did not reduce basal progesterone secretion suggesting involvement of gene transcription and translation in the GH and IGF-I action on luteal cells. Additionally, the activity of 3ß-hydroxysteroid dehydrogenase (3ß-HSD) under the influence of GH added alone or together with IGF was measured by the conversion of pregnenolone to progesterone. Stimulation of P4 secretion in P5-treated cells in GH-stimulated cultures was not observed, however, high stimulatory effect was noted in IGF-I treated cultures. In conclusion, the present studies indicate that there is direct and cycle stage dependent influence of GH and IGF-I on steroidogenesis in porcine luteal cells. It is suggested that both IGF and GH may exert some regulatory action during CL development in the pig