Ovarian steroid treatment decreases corticotropin-releasing hormone (CRH) mRNA and protein in the hypothalamic paraventricular nucleus of ovariectomized monkeys

Corticotropin-releasing hormone (CRH) gene and protein expression were examined in the paraventricular nucleus (PVN) of ovariectomized female macaques treated with placebo or hormone therapy (HT) consisting of either estrogen (E) for 28 days, or progesterone (P) for the last 14 of 28 days, or E for 28 days supplemented with P for the last 14 of 28 days using Silastic capsules implanted s.c. in the periscapular region (n=4/group). Perfusion fixed sections (25 μm) at five levels of the PVN (rostral to caudal at 250 μm intervals) were immunostained (ICC) with an antibody to human CRH or processed in an in situ hybridization (ISH) assay with a monkey specific CRH riboprobe. The immunostained CRH-positive area was quantified with a Marianas Stereology Workstation and Slidebook 4.2. There was a significant decrease in the immunological CRH signal with E, P, and E+P treatment as measured by total or average pixels and microns (analysis of variance (ANOVA), p<0.002; Student-Newman-Keul's post hoc test versus placebo control group, p<0.05). There was also a decrease in the number of detectable CRH neurons (ANOVA, p<0.03) with HT. The sections processed for ISH were exposed to autoradiographic films. The CRH mRNA signal was analyzed with NIH Image. The average optical density and positive pixel area of the CRH mRNA signal was significantly suppressed by ovarian HT (ANOVA p<0.002; Student-Newman-Keul's post hoc test versus placebo control group, p<0.05). In summary, 1 month of stable treatment with a moderate dose of E, P or E+P significantly reduced CRH mRNA and protein in the PVN of ovariectomized monkeys. These results suggest that this hormone treatment regimen may increase stress resilience in surgically menopausal primates.