Distribution and effects of testosterone on aromatase mRNA in the quail forebrain: a non-radioactive in situ hybridization study

Abstract

A number of studies have been devoted to the analysis of the anatomical distribution, control by steroids and functional significance of aromatase (the enzyme metabolizing testosterone into 17beta-estradiol) in the quail brain. In particular, the sexually dimorphic nucleus preopticus medialis has been the main focus of investigation because testosterone aromatization in this structure mediates the activation of male sexual behavior and aromatase activity is itself testosterone-dependent in this nucleus. No information on the anatomical distribution of aromatase gene expression is, however, available so far in this avian species. In the present study we applied a non-radioactive in situ hybridization technique to describe the distribution of aromatase mRNA containing neurons in the quail prosencephalon. We also analyzed, at a neuronal level of resolution, the induction by testosterone of this mRNA in the medial preoptic nucleus. Dense clusters of aromatase gene expressing neurons were observed within the medial preoptic nucleus, the nucleus of the stria terminalis, the ventro-medial hypothalamus and the tuberal region. Scattered neurons expressing lower levels of aromatase mRNA were also found in the dorsal thalamic area and central gray. The specificity of the staining was confirmed by demonstrating the absence of signal in sections that had been hybridized with a sense probe. Moreover, the distribution of the aromatase mRNA containing cells completely overlapped with the distribution of the aromatase-immunoreactive cells. Aromatase-mRNA expression was controlled by testosterone (or its metabolites) in the entire medial preoptic nucleus. Castration resulted in a decrease in the number of aromatase mRNA-containing cells and this effect was totally reversed by testosterone treatment. These data further support the idea that testosterone regulates the rate of its own aromatization by modulating the expression of aromatase rather than by acting at a post transcriptional level.