Urbatzka R, Lutz I, Kloas W
Department of Inland Fisheries, Leibniz-Institute of Freshwater Ecology and Inland Fisheries, Müggelseedamm 310, 12587 Berlin, Germany.
Gen Comp Endocrinol. 2007 Aug-Sep;153(1-3):280-8. doi: 10.1016/j.ygcen.2007.01.041. Epub 2007 Feb 9.
The key enzymes involved in the production of endogenous sex steroids are steroid-5-alpha-reductase and aromatase converting testosterone (T) into dihydrotestosterone (DHT) and into estradiol (E2), respectively. To gain more insights into the molecular mechanisms of sexual differentiation of amphibians, we determined the mRNA expression of steroid-5-alpha-reductase type1 (Srd5a1), type2 (Srd5a2) and aromatase (Aro) during ontogeny starting from the egg and ending after completion of metamorphosis in Xenopus laevis. Expression of all three enzymes was measured by means of semi-quantitative RT-PCR, determining for the first time Srd5a1 and Srd5a2 mRNA expression in amphibians. mRNA was analyzed in whole body homogenates from stage 12 to 48, while brain and gonads with kidney were studied separately from stage 48 to 66. Different ontogenetic mRNA expression patterns were observed for all genes analyzed, revealing early mRNA expression of Srd5a1 already in the egg at stage 12 whereas Srd5a2 and Aro was detected at stage 39. Sex-specific mRNA expressions of Srd5a2 and of Aro were determined in the gonads with kidney but not in brain. Srd5a2 was two-fold higher expressed in testes than in ovaries while Aro mRNA was ten-fold higher in ovaries. No gender-specific mRNA expression was observed for Srd5a1 in gonads and in brain. The ontogenetic patterns of Aro, Srd5a1 and Srd5a2 suggest that these genes are involved in sexual differentiation of gonads and brain already in early developmental stages. Especially in gonads Srd5a2 seems to be important for physiological regulation of testis development while Aro is associated with the development of ovaries.
参与内源性性类固醇生成的关键酶是类固醇5-α-还原酶和芳香化酶,它们分别将睾酮(T)转化为二氢睾酮(DHT)和雌二醇(E2)。为了更深入了解两栖动物性别分化的分子机制,我们测定了非洲爪蟾从卵开始到变态完成后的个体发育过程中类固醇5-α-还原酶1型(Srd5a1)、2型(Srd5a2)和芳香化酶(Aro)的mRNA表达。通过半定量RT-PCR测定了这三种酶的表达,首次确定了两栖动物中Srd5a1和Srd5a2的mRNA表达。在第12至48阶段的全身匀浆中分析mRNA,而在第48至66阶段分别研究脑、性腺和肾脏。在所分析的所有基因中观察到不同的个体发育mRNA表达模式,显示Srd5a1在第12阶段的卵中就已出现早期mRNA表达,而Srd5a2和Aro在第39阶段被检测到。在性腺和肾脏中测定了Srd5a2和Aro的性别特异性mRNA表达,但在脑中未检测到。Srd5a2在睾丸中的表达比在卵巢中高两倍,而Aro mRNA在卵巢中的表达高十倍。在性腺和脑中未观察到Srd5a1的性别特异性mRNA表达。Aro、Srd5a1和Srd5a2的个体发育模式表明,这些基因在早期发育阶段就参与性腺和脑的性别分化。特别是在性腺中,Srd5a2似乎对睾丸发育的生理调节很重要,而Aro与卵巢的发育有关。