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Expression of StAR and Key Genes Regulating Cortisol Biosynthesis in Near Term Ovine Fetal Adrenocortical Cells: Effects of Long-Term Hypoxia

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Abstract

We previously demonstrated decreased expression of key genes regulating Cortisol biosynthesis in long-term hypoxic (LTH) sheep fetal adrenals compared to controls. We also showed that inhibition of the extracellular signal-regulated kinases (ERKs) with the mitogen-activated protein kinase (MEK)/ERK inhibitor UO126 limited adrenocorticotropic (ACTH)-induced Cortisol production in ovine fetal adrenocortical cells (FACs), suggesting a role for ERKs in Cortisol synthesis. This study was designed to determine whether the previously observed decrease in LTH cytochrome P4501 1A1/cytochrome P450c17 (CYP1 1A1/CYP17) in adrenal glands was maintained in vitro, and whether ACTH alone with or without UO126 treatment had altered the expression of CYP1 1A1, CYP17, and steroidogenic acute regulatory protein (StAR) in control versus LTH FACs. Ewes were maintained at high altitude (3820 m) from ≈40 days of gestation (dG). At 138 to 141 dG, fetal adrenal glands were collected from LTH (n = 5) and age-matched normoxic controls (n = 6). Fetal adrenocortical cells were challenged with ACTH (10−8 M) with or without UOI26 (10 (J.M) for 18 hours. Media samples were collected for Cortisol analysis and messenger RNA (mRNA) for CYP1 1A1, CYP1 7, and StAR was quantified by quantitative real-time polymerase chain reaction. Cortisol was higher in the LTH versus control (P <.05). StAR mRNA was decreased in LTH versus control (P <.05). U0126 alone had no effect on mRNA in either group. UO126 prevented the increase in CYP1 1A1 and CYP17 in control FACs. Basal CYP1 1A1 and CYP1 7 were not different in LTH versus control. ACTH increased CYP1 1A1 and CYP17 only in control FACs (P <.05). U1026 attenuated the ACTH response indicative of a role for ERK in CYP1 1A1 and CYP17 expression. ACTH may require additional factors in FACs to fully regulate StAR expression.

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Authors and Affiliations

  1. Center for Perinatal Biology, Loma Linda University, Loma Linda School of Medicine, CA, 92350, USA

    Vladimir E. Vargas PhD, Kanchan M. Kaushal MS & Charles A. Ducsay PhD

  2. Department of Obstetrics and Gynecology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Vladimir E. Vargas PhD & Dean A. Myers PhD

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  1. Vladimir E. Vargas PhD
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  2. Dean A. Myers PhD
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  3. Kanchan M. Kaushal MS
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  4. Charles A. Ducsay PhD
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Correspondence to Charles A. Ducsay PhD.

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Vargas, V.E., Myers, D.A., Kaushal, K.M. et al. Expression of StAR and Key Genes Regulating Cortisol Biosynthesis in Near Term Ovine Fetal Adrenocortical Cells: Effects of Long-Term Hypoxia. Reprod. Sci. 25, 230–238 (2018). https://doi.org/10.1177/1933719117707056

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