Reproductive and Molecular Biology Group, Department of Structural and Functional Biology, Institute of Biosciences, São Paulo State University, Botucatu, São Paulo, Brazil.
Department of Animal Morphology and Physiology, Faculty of Agricultural and Veterinary Sciences, São Paulo State University, Jaboticabal, São Paulo, Brazil; Aquaculture Center (CAUNESP), São Paulo State University, Jaboticabal, São Paulo, Brazil.
Comp Biochem Physiol B Biochem Mol Biol. 2020 Oct-Nov;248-249:110467. doi: 10.1016/j.cbpb.2020.110467. Epub 2020 Jul 3.
Gonadal sex differentiation in teleost fish shows greater plasticity as compared to other vertebrates, as it can be influenced by a variety of factors such as exogenous sex steroids. Exogenous estrogens, such as 17β-estradiol (E2), can induce feminization when administered during early embryonic development. However, the mechanisms underlying the E2-induced feminization are not fully understood, especially in Neotropical species. Therefore, the aim of this study was to evaluate the effects of E2 administration on the phenotypic sex characteristics, histological assessment of the gonads, and the expression of selected genes in Astyanax altiparanae exposed to dietary E2 prior to gonadal differentiation. At 4 days post-hatch (dph), groups of 30-40 undifferentiated larvae were fed with a diet containing varying amounts of E2 for 28 days, and fish were sampled at 90 dph. Previous studies revealed that ovary formation in A. altiparanae occurred at 58 dph, whereas the first sign of testis formation was found at 73 dph. In relation to the control, E2 exposure increased the proportion of phenotypic females in 120% and 148.4% for 4 and 6 mg E2/Kg, respectively. However, histological analysis revealed that treatments did not affect gonadal sex ratio between males and females, but induced intersex (testis-ova) in the group treated with 6 mg E2/Kg food. Treatment with E2 also altered gonadal transcript levels of a selected number of genes implicated in sexual differentiation. Males overexpressed dmrt1, sox9 and amh following E2 treatment as compared to control. Females showed increased mRNA levels of dmrt1 and sox9, which might be related to the down-regulation of cyp19a1a after E2 exposure. In summary, E2 exposure during early gonadal development affected male secondary characteristics without changing the gonadal sex ratio, and altered expression of genes implicated in sexual differentiation.
鱼类的性腺性别分化比其他脊椎动物具有更大的可塑性,因为它可以受到各种因素的影响,如外源性性激素。外源性雌激素,如 17β-雌二醇(E2),在胚胎早期发育时给药会诱导雌性化。然而,E2 诱导雌性化的机制尚不完全清楚,特别是在新热带物种中。因此,本研究旨在评估 E2 给药对暴露于 E2 饮食前性腺分化的 Astyanax altiparanae 的表型性别特征、性腺组织学评估和选定基因表达的影响。在孵化后 4 天(dph),将 30-40 个未分化的幼虫分组,用含有不同量 E2 的饮食喂养 28 天,然后在 90 dph 取样。先前的研究表明,卵巢在 A. altiparanae 中形成于 58 dph,而睾丸形成的第一个迹象是在 73 dph 发现的。与对照组相比,E2 暴露分别使 4 和 6 mg E2/Kg 组的表型雌性比例增加了 120%和 148.4%。然而,组织学分析表明,处理并未影响雄性和雌性之间的性腺性别比例,但在 6 mg E2/Kg 食物处理组中诱导了间性(睾丸-卵)。E2 处理还改变了选定数量与性分化相关的基因在性腺中的转录水平。与对照组相比,E2 处理后雄性 dmrt1、sox9 和 amh 过度表达。雌性 dmrt1 和 sox9 的 mRNA 水平增加,这可能与 E2 暴露后 cyp19a1a 的下调有关。总之,早期性腺发育过程中 E2 的暴露影响了雄性的次级特征,而不改变性腺性别比例,并改变了与性分化相关的基因的表达。