Vargas VE, Kaushal KM, Monau TR, Myers DA, Ducsay CA. Extracellular signal-regulated kinases (ERK1/2) signaling pathway plays a role in cortisol secretion in the long-term hypoxic ovine fetal adrenal near term. Am J Physiol Regul Integr Comp Physiol 304: R636 -R643, 2013. First published February 20, 2013 doi:10.1152/ajpregu.00318.2012.-This study assessed the role of the extracellular signal-regulated kinase (ERK) signaling pathway on the previously observed enhanced cortisol secretion in response to adrenocorticotropic hormone (ACTH) treatment in fetal adrenocortical cells (FACs) from long-term hypoxic (LTH) ovine fetuses. Ewes were maintained at high altitude (3,820 m) from ϳ40 to 138 -141 days gestation when FACs were collected and challenged with either ACTH (10 nM) or 8-bromoadenosine 3=,5=-cyclic monophosphate (8-bromo-cAMP, 10 mM) in the presence or absence of the mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MEK)/ERK inhibitor UO126 (10 M). FACs from age-matched normoxic fetuses served as controls. Media and FACs were collected at selected time intervals after ACTH or 8-bromocAMP stimulation for cortisol measurement and Western analysis of ERK1/2 and phospho-ERK1 and -2 (pERK1/2). After ACTH or 8-bromo-cAMP treatment, cortisol production was greater in the LTH group compared with control (P Ͻ 0.05). UO126 reduced ACTH and 8-bromo-cAMP-mediated cortisol output in both groups (P Ͻ 0.01 vs. ACTH or 8-bromo-cAMP alone). Under basal conditions, ERK1/2 and pERK1/2 were not different between LTH and normoxic fetuses. In response to ACTH or 8-bromo-cAMP treatment, ERK1/2 were not different between groups; however, pERK1/2 were elevated in the LTH FACs compared with normoxic control FACs. ERK1/2 phosphorylation declined following ACTH treatment in the control group, but UO126 had no effect on ERK1/2 compared with untreated levels. Both ACTH and 8-bromo-cAMP treatment resulted in a decline of protein levels. UO126 pretreatment virtually eliminated pERK1/2 expression. We conclude that basal ERK signaling in FACs is necessary for normal cortisol production and sustained pERK in LTH adrenals enhances cortisol production.ACTH; UO126; 8-bromo-cAMP; sheep; fetus THE CLASSICAL adrenocorticotropic hormone (ACTH) signaling pathway via 3,5-cAMP is generally accepted as the major mechanism regulating cortisol biosynthesis in the adrenal cortex (10, 36). The late gestation increase in fetal plasma cortisol resulting from adrenocortical maturation is critical for optimal fetal organ maturation, and in sheep, it plays an essential role in parturition (24,25). Hypoxia has clearly been shown to play a key role in the regulation of the fetal hypothalamic-pituitaryadrenal axis (3,17,11). Importantly, from a clinical perspective, not only women that live at high altitude, but also women that smoke, have heart/lung disease, or are anemic during pregnancy expose the fetus to hypoxia (41, 22, 31).Our laboratory has developed an ovine model of high altitude-induced long-term hypoxia (LTH). We have fo...