Preliminary array analysis reveals novel genes regulated by ovarian steroids in the monkey raphe region

We hypothesize that ovarian hormones may improve serotonin neuron survival. We sought the effect of estradiol (E) and progesterone (P) on novel gene expression in the macaque dorsal raphe region with Affymetrix array analysis. Nine spayed rhesus macaques were treated with either placebo, E or E+P via Silastic implant for 1 month prior to euthanasia (n=3 per treatment). RNA was extracted from a small block of midbrain containing the dorsal raphe and examined on an Agilent Bioanalyzer. The RNA from each monkey was labeled and hybridized to an Affymetrix HG_U95AV Human GeneChip Array. After filtering and sorting, 25 named genes remained that were regulated by E, and 24 named genes remained that were regulated by supplemental P. These genes further sorted into functional categories that would promote neuronal plasticity, transmitter synthesis, and trafficking, as well as reduce apoptosis. The relative abundance of four pivotal genes was examined in all nine animals with quantitative RT-PCR and normalized by glyceraldehyde 3-phosphate dehydrogenase (GAPDH). E±P caused a significant threefold reduction in JNK-1 (a pro-apoptosis gene, p<0.007); and a significant sixfold decrease in kynurenine mono-oxygenase (produces neurotoxic quinolones, p<0.05). GABA-A receptor (α3 subunit; benzodiazepine site) and E2F1 (interferes with cytokine signaling) were unaffected by E, but increased sevenfold (p<0.02) and fourfold (p<0.009), respectively, upon treatment with P. In summary, subsets of genes related to tissue remodeling or apoptosis were up- or down-regulated by E and P in a tissue block containing the dorsal raphe. These changes could promote cellular resilience in the region where serotonin neurons originate.