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Parturition and fetal organ maturation in sheep are associated with increased activity of the fetal hypothalamic-pituitary-adrenal (HPA) axis during late pregnancy. However, the factors responsible for HPA activation remain unclear. In the fetal pituitary, levels of pro-opiomelanocortin (POMC) mRNA increase, but the numbers of binding sites for corticotrophin-releasing hormone (CRH), and ACTH responsiveness to exogenous CRH decline during the last 20 days of pregnancy. We have examined regulation of CRH binding, pituitary ACTH responsiveness, and levels of POMC mRNA in cultures of adenohypophysial cells from term fetal sheep. After a 4-day stabilization period, output of immunoreactive (ir) ACTH was increased over 48 h in a dose-dependent fashion by both CRH and arginine vasopressin (AVP) but decreased by cortisol. Subsequent output of ir-ACTH to a 3-h challenge with 100 nM CRH was attenuated after pretreatments with CRH, AVP or cortisol; the effect of CRH being greater than that of cortisol or AVP. At the end of 48 h of treatment with CRH, AVP or cortisol, there was a 40–50% reduction in the number of CRH-binding sites, but the levels of POMC mRNA decreased significantly only after cortisol treatment and were not altered significantly by CRH or AVP.
We conclude that under the conditions of these experiments, CRH and AVP increase ir-ACTH output without increasing the level of steady-state POMC mRNA, but may contribute to loss of pituitary responsiveness to CRH by down-regulation of CRH receptor number. Cortisol exerts negative feedback on POMC mRNA and decreases the number of CRH receptors. Thus, any one or all of CRH, AVP and cortisol could be responsible for the decline in CRH binding in the fetal sheep pituitary during late pregnancy. Although CRH and AVP may affect secretion of ir-ACTH, the present results do not support a role for these neuropeptides in affecting the level of POMC mRNA in the fetal sheep pituitary.
Journal of Endocrinology (1994) 143, 199–208
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