Receptive behavior in females vertebrates is controlled by hormones, principally estrogen, secreted by the ovary. Estrogen influences behavior by interacting with a specific estrogen binding protein, or receptor, located in target cells in certain hypothalamic nuclei. To better understand the molecular mechanisms involved in the control of receptive behavior in whiptail lizards, we investigated the effects of exogenous estrogen on the regulation of estrogen receptor and progesterone receptor expression in several regions of the brains of Cnemidophorus uniparens. First we determined a dosage of 17 beta-estradiol 3-benzoate (0.5 micrograms) which reliably induced receptive behavior in ovariectomized C. uniparens. Then using in situ hybridization, we examined the effects of that dosage on the expression of estrogen receptor and progesterone receptor mRNA in the brain 24 h after injection. Estrogen treatment resulted in a significant up-regulation of estrogen receptor mRNA expression in the ventromedial nucleus of the hypothalamus and torus semicircularis, down-regulation of estrogen receptor mRNA expression in the lateral septum, and no change in the periventricular nuclei of the hypothalamus, the periventricular nucleus of the preoptic area, and the dorsal hypothalamus. The same dosage resulted in increased progesterone receptor mRNA expression in the ventromedial nucleus of the hypothalamus and the periventricular nucleus of the preoptic area; no significant changes in progesterone receptor mRNA expression were observed in the periventricular nuclei of the hypothalamus or the torus semicircularis, although the differences in progesterone receptor expression in the torus semicircularis approached statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS)

Circulating concentrations of gonadal steroid hormones and reproductive behavior in female vertebrates vary as a function of ovarian state. Steroids secreted by the ovary, specifically estrogen and progesterone, influence the expression of behaviors associated with reproduction by intracellular sex steroid receptors located in specific regions of the brain. Using in situ hybridization, we analyzed estrogen receptor and progesterone receptor messenger RNA expression in several brain regions of ovariectomized, vitellogenic, and postovulatory individuals from two species of whiptail lizards (Cnemidophorus uniparens and C. inornatus). Although these species are genetically very similar, they differ in two aspects of their reproductive biology: (i) the unisexual C. uniparens alternate between expressing female-typical and male-like pseudosexual behaviors while female C. inornatus normally express only female receptive behavior, and (ii) circulating estradiol concentrations in reproductively active female C. uniparens are approximately five-fold lower than in reproductively active female C. inornatus. We found that the regulation of sex steroid receptor gene expression was region specific, with receptor-mRNA expression being increased, unchanged, or decreased during vitellogenesis depending on the area. Furthermore, several species differences in the amount of sex steroid receptor-mRNA were found that may be relevant to the species differences in circulating estrogen concentrations and sexual behavior.

Cnemidophorus uniparens is a unisexual species of whiptail lizard of hybrid origin whereas C. inornatus is a sexual species and the maternal ancestor of C. uniparens. Together they represent an excellent model system for investigating the evolution of hormone-brain-behavior relationships. Normal circulating estradiol (E) concentrations in C. uniparens are approximately 5-fold lower than those of female C. inornatus in a similar reproductive state. Experiments were performed to determine whether (i) C. uniparens is more sensitive to E, and (ii) whether the difference in sensitivity is correlated with differences in estrogen receptor (ER)-mRNA expression in the brain. Dose-response curves reveal that ovariectomized C. uniparens are more responsive than ovariectomized C. inornatus to exogenous estradiol 17β-benzoate (EB). EB is more effective in C. uniparens at inducing receptive behavior and progesterone receptor (PR) gene expression in the ventromedial nucleus of the hypo-thalamus (VMH). In situ hybridization analysis of ER-mRNA expression revealed no species differences in ER-mRNA content in the VMH of ovariectomized animals. Treatment of ovariectomized animals with EB resulted in a greater induction of ER-mRNA expression in the VMH of C. uniparens compared to C. inornatus. These results indicate that the differences in behavioral sensitivity to E lie in the estrogen target neurons in the brain region controlling receptive behavior, the VMH, and that the difference in sensitivity cannot be explained by species differences in the basal rate of ER gene expression.

Sex and species differences in androgenic regulation of steroid hormone receptor mRNAs were examined in the diencephalon of two species of whiptail lizards: Cnemidophorus inornatus is a sexual species and the direct evolutionary ancestor to Cnemidophorus uniparens, an all-female parthenogenetic species. Lizards were gonadectomized and treated with different doses of either aromatizable testosterone or nonaromatizable dihydrotestosterone. The relative abundances of androgen-, oestrogen-, and progesterone-receptor mRNAs were compared in various nuclei following in situ hybridization with homologous riboprobes. A diversity of patterns in androgenic regulation was observed, with effects differing according to brain region, the steroid-receptor mRNA being considered and, in some cases, between androgens. In the ancestral sexual species, intact males had lower androgen-receptor mRNA abundances than castrated, blank-implanted males in the medial preoptic area. Testosterone significantly decreased androgen-receptor mRNA abundance in the medial preoptic area of castrated males. Males had higher androgen-receptor mRNA levels in the preoptic area than females generally and neither the sexual or parthenogenetic females showed a decrease in androgen-receptor mRNA with androgen treatment. Both testosterone and dihydrotestosterone increased oestrogen-receptor mRNA abundance in the ventromedial hypothalamus of C. inornatus, but no sex differences in this effect were observed. Gonadectomy decreased, whereas androgen treatment increased, progesterone-receptor mRNA abundance in the ventromedial hypothalamus. There was a sex difference in this response to androgen in the sexual species, with males having greater amounts than females in this brain area. The parthenogenetic species exhibited a similar pattern to females of the sexual species, but the levels were higher overall, possibly because Cnemidophorus uniparens is triploid. The periventricular preoptic area showed a different pattern, with testosterone treatment increasing progesterone-receptor mRNA abundance in both sexes of the sexual species and in the parthenogenetic species, while dihydrotestosterone did not. The diversity of patterns in androgen effects indicates that gonadal sex, aromatization of androgen, and perhaps gene dosage all influence the expression of steroid-receptor mRNAs in the lizard brain.

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