Diminished expression of ACTH signaling proteins and steroidogenic limiting factors in adrenocortical cells isolated from halothanenn pigs
Li, Liu-An | Xia, Dong | Wei, Shi | Li, Xiao | Parvizi, Nahid | Zhao, Ru-Qian
Previous studies demonstrated significantly lower plasma cortisol level in homozygous halothane-positive (Halnn) pigs, as compared with homozygous halothane-negative (HalNN) pigs. To determine whether such difference is attributed to the fundamental alterations in adrenocortical function, F1 offsprings from Pietrain (Halnn)xErhualian (HalNN) were intercrossed to produce F₂ sibling pigs with segregated genotypes. Adrenocortical cells were isolated from the Halnn and HalNN F₂ pigs, respectively, and cultured with or without ACTH challenge. Cortisol levels in culture medium, as well as the content of MC2R, cAMP, CREB, phosphorylated CREB (pCREB), StAR and P450scc in adrenocortical cell lysates, were determined. Cortisol, cAMP, StAR and P450scc levels were significantly lower in Halnn adrenocortical cells under basal condition without ACTH challenge. ACTH significantly increased cortisol level in the medium and the protein content of MC2R, StAR, P450scc in adrenocortical cell lysates, regardless of genotypes. Total CREB protein content was not different between genotypes and treatments, whereas pCREB content exhibited significant effects of genotype and treatment, being higher in HalNN than in Halnn under basal condition and in response to ACTH challenge. These results indicate that the compromised cAMP/PKA/pCREB signaling pathway of ACTH and diminished expression of limiting factors in adrenocortical steroidogenesis (StAR and P450scc) may contribute to the significantly lower plasma cortisol levels in Halnn pigs.
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