Department of Animal Sciences, School of Life Sciences-Centre for Advanced Studies, University of Hyderabad, P.O. Central University, Hyderabad-500 046, Andhra Pradesh, India.
Steroids. 2012 Sep;77(11):1133-40. doi: 10.1016/j.steroids.2012.06.002. Epub 2012 Jul 11.
3β-hydroxysteroid dehydrogenase (3β-hsd) plays an important role in biosynthesis of both androgens and estrogens during steroidogenesis. In this study, we report the cloning of a full-length cDNA of 3β-hsd from gonads of the air-breathing catfish, Clarias gariepinus a seasonally reproducing teleost fish. We studied the expression pattern of 3β-hsd during gonadal ontogeny and recrudescence (flanking two years of reproductive cycle) using real-time PCR. We also examined the influence of gonadotropin on 3β-hsd expression in gonads of catfish by human chorionic gonadotropin (hCG) induction. The real-time PCR results revealed that 3β-hsd transcript was detectable much earlier in undifferentiated gonads i.e. before the sex differentiation and later on its expression was seen in both male and female gonads throughout the development. The expression analysis during subsequent seasonal reproductive cycle in catfish (older than one year) showed that in adult males, the transcripts were significantly high during prespawning phase (spermatogenesis) and declined during spermiation. In adult females, the transcripts were abundantly expressed in the ovarian follicles both at prespawning and spawning phases. Furthermore, the 3β-hsd mRNA levels in different follicular stages were markedly high in vitellogenic follicles (maturing oocytes; stage III) compared to other stages. Treatment of hCG in recrudescing female fish, in vivo as well as in testicular slices, in vitro resulted in the up-regulation of gonadal 3β-hsd mRNA indicating that it is under the regulation of gonadotropins. These results together suggest that 3β-hsd gene plays an important role during spermatogenesis and oogenesis as well as in the gonadal recrudescence of catfish.
3β-羟甾脱氢酶(3β-hsd)在类固醇生成过程中对于雄激素和雌激素的生物合成起着重要作用。本研究报告了从具有季节性繁殖的硬骨鱼胡子鲶的性腺中克隆全长 cDNA 的情况。我们使用实时 PCR 研究了 3β-hsd 在性腺发生和重新发育过程中的表达模式(该过程围绕两个生殖周期进行)。我们还通过人绒毛膜促性腺激素(hCG)诱导检查了促性腺激素对胡子鲶性腺中 3β-hsd 表达的影响。实时 PCR 结果表明,3β-hsd 转录本在未分化的性腺中即可检测到,即性别分化之前,随后在整个发育过程中都可在雄性和雌性性腺中检测到其表达。在鱼的随后的季节性生殖周期(一年以上)中的表达分析表明,在成年雄性中,在产卵前期(精子发生)时转录本显著升高,而在精子排放时则下降。在成年雌性中,在产卵前期和产卵期,卵巢滤泡中的转录本均大量表达。此外,3β-hsd mRNA 水平在不同的滤泡阶段中在卵黄发生滤泡(成熟卵母细胞;第 III 期)中明显高于其他阶段。在体内和体外的再生雌性鱼以及睾丸切片中用 hCG 处理可导致性腺 3β-hsd mRNA 的上调,表明其受促性腺激素的调节。这些结果共同表明,3β-hsd 基因在精子发生和卵子发生以及胡子鲶的性腺重新发育过程中起着重要作用。