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Sexually differentiated and neuroanatomically specific co‐expression of aromatase neurons and GAD67 in the male and female quail brain

Testosterone aromatization into estrogens in the preoptic area (POA) is critical for the activation of male sexual behavior in many vertebrates. Yet, the cellular mechanisms mediating actions of neuroestrogens on sexual behavior remain largely unknown. We investigated in male and female Japanese quail by dual‐label fluorescent in situ hybridization (FISH) whether aromatase‐positive (ARO) neurons express glutamic acid decarboxylase 67 (GAD67), the rate‐limiting enzyme in GABA biosynthesis. ARO cells and ARO cells double labeled with GAD67 (ARO‐GAD67) were counted at standardized locations in the medial preoptic nucleus (POM) and the medial bed nucleus of the stria terminalis (BST) to produce three‐dimensional distribution maps. Overall, males had more ARO cells than females in POM and BST. The number of double‐labeled ARO‐GAD67 cells was also higher in males than in females and greatly varied as a function of the specific position in these nuclei. Significant sex differences were however present only in the most caudal part of POM. Although both ARO and GAD67 were expressed in the VMN, no colocalization between these markers was detected. Together, these data show that a high proportion of estrogen‐synthesizing neurons in POM and BST are inhibitory and the colocalization of GAD67 with ARO exhibits a high degree of anatomical specificity as well as localized sex differences. The fact that many preoptic ARO neurons project to the periaqueductal gray in male quail suggests possible mechanisms through which locally produced estrogens could activate male sexual behavior.