Regulation of adrenocorticotropin secretion in vitro by anterior pituitary corticotrophs from fallow deer (Dama dama).
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abstract
The actions of corticotropin-releasing hormone (CRH), vasopressin (VP), the synthetic glucocorticoid dexamethasone (DEX), and mifepristone (RU 486), a glucocorticoid antagonist, on the secretion of adrenocorticotropin (ACTH) by cultured fallow deer corticotrophs were studied in vitro. On Day 5 of primary culture, corticotrophs were challenged for up to 4 hr with medium alone (Control), CRH, VP, DEX, forskolin (FSK), phorbol ester (TPA), cyclic AMP (cAMP), and/or RU 486 at various concentrations and combinations. CRH, VP, FSK and TPA each stimulated (P < 0.01) the secretion of ACTH in dose- and time-related manners. Relative to Control, CRH at 0.001 and 0.1 microM and VP at 0.01 and 1 microM increased (P < 0.01) medium concentration of ACTH by 7.3-, 13.5-, 3.7- and 9.0-fold, respectively. There was a treatment x incubation time interaction (P < 0.01) such that at 30-min posttreatment, CRH-induced ACTH secretion tended (P < 0.10) to be less than that obtained via VP treatment, whereas at 1, 3, and 4 hr posttreatment, medium concentration of ACTH from cells treated with 0.1 microM CRH was greater (P < 0.05) than that in cells treated with 1 microM VP. At equimolar doses of 0.01 and 0.1 microM, CRH was 3.4- and 3.0-fold more potent (treatment x dose, P < 0.05) than VP. Cotreatment with 1 microM DEX reduced (P < 0.001) the stimulatory effects of CRH (0.1 microM), VP (1 microM), FSK (10 microMs), TPA (0.1 microM), and cAMP (0.001 M). However, the coaddition of RU 486 (1 microM) to the CRH plus DEX- and the FSK plus DEX-treated wells partially negated the inhibitory effects of DEX. RU 486 completely negated the inhibitory effects of DEX on the VP-, TPA-, and cAMP-stimulated secretion of ACTH. These data indicate that CRH is a more potent stimulator of ACTH secretion than is VP in primary culture of fallow deer pituitary cells. This study also demonstrates the utility of an in vitro culture system to investigate stress-related hormonal interactions in cervids.
