Zou Congcong, Wang Wenxiang, Shu Chang, Liang Shaoshuai, Zou Yuxia, Wang Lijuan, Wu Zhihao, Liu Yan, You Feng
Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China; Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China; Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2023 Jun-Jul;266:110848. doi: 10.1016/j.cbpb.2023.110848. Epub 2023 Mar 16.
Steroidogenesis is an important biological process for gonadal differentiation and development. In mammals, 3β-hydroxysteroid dehydrogenase 7 (HSD3B7) could convert 3β-hydroxy of 7α-hydroxycholesterol into a ketone and form 7α-hydroxy-4-cholesten-3-one, which may affect steroidogenesis. However, in fish, the study of Hsd3b7 is still lacking. In this study, Hsd3b7 was identified in the olive flounder Paralichthys olivaceus, an important mariculture fish. According to bioinformatics analysis, Hsd3b7 belongs to a Rossmann-fold NAD(P)(+)-binding protein and can interact in a predictable manner with Hsd17b2, -3, and - 4, which play a role in steroidogenesis. In the adult flounder, Hsd3b7 was expressed in various tissues, at particularly high level in male muscle. The expression levels of Hsd3b7 at gonadal development stages I-V initially increased and then decreased, with an inflection point in the ovary at stage III and in the testis at stage IV. At stage III, the expression level of Hsd3b7 was significantly higher in the ovary than in the testis (P < 0.01). The results of in situ hybridization (ISH) revealed that it was mainly expressed in oocytes of phases I-IV or around oocytes of phases IV-V in the ovaries and around spermatid lobules at stages IV-V in the testes. Three regulatory sites of SRY-box transcription factor 9 (Sox9), a transcription factor involved in steroidogenesis and gonadal differentiation, were predicted in the promoter of Hsd3b7. After intraperitoneal injection with the recombination flounder Sox9a, the expression of Hsd3b7 was significantly up-regulated (P < 0.01). During the flounder gonadal differentiation, 17β-estradiol (E2, 5 μg/g feed) and 17α-methyltestosterone (T, 5 μg/g feed) were used to obtain the phenotypic female or male flounder, and the results showed that in the E2 group, Hsd3b7 expression was highest at 2 cm TL, the primordial gonad stage, which was significantly higher than that at 12 cm TL (P < 0.05). In the T group, Hsd3b7 expression level was also highest at 2 cm TL and significantly higher than at 10 and 12 cm TL (P < 0.05). Moreover, Hsd3b7 was detected to be localized mainly around oogonia and spermatogonia during the differentiated gonads with ISH. These findings first introduce the expression characteristics of Hsd3b7 and the effect of Sox9a on its expression, which contribute to our understanding of the function of Hsd3b7 in fish gonads.
类固醇生成是性腺分化和发育的重要生物学过程。在哺乳动物中,3β-羟基类固醇脱氢酶7(HSD3B7)可将7α-羟基胆固醇的3β-羟基转化为酮,形成7α-羟基-4-胆甾烯-3-酮,这可能影响类固醇生成。然而,在鱼类中,对Hsd3b7的研究仍然缺乏。在本研究中,在重要的海水养殖鱼类褐牙鲆(Paralichthys olivaceus)中鉴定出了Hsd3b7。根据生物信息学分析,Hsd3b7属于Rossmann折叠NAD(P)(+)结合蛋白,并且可以与在类固醇生成中起作用的Hsd17b2、-3和-4以可预测的方式相互作用。在成年牙鲆中,Hsd3b7在各种组织中表达,在雄性肌肉中的表达水平特别高。Hsd3b7在性腺发育I-V期的表达水平最初升高然后降低,在卵巢中III期和睾丸中IV期出现拐点。在III期,卵巢中Hsd3b7的表达水平显著高于睾丸(P < 0.01)。原位杂交(ISH)结果显示,它主要在卵巢中I-IV期的卵母细胞或IV-V期卵母细胞周围以及睾丸中IV-V期的精小叶周围表达。在Hsd3b7的启动子中预测到了参与类固醇生成和性腺分化的转录因子SRY-box转录因子9(Sox9)的三个调控位点。腹腔注射重组牙鲆Sox9a后,Hsd3b7的表达显著上调(P < 0.01)。在牙鲆性腺分化期间,使用17β-雌二醇(E2,5 μg/g饲料)和17α-甲基睾酮(T,5 μg/g饲料)获得表型雌性或雄性牙鲆,结果表明,在E2组中,Hsd3b7在2 cm体长(原基性腺期)时表达最高,显著高于12 cm体长时的表达(P < 0.05)。在T组中,Hsd3b7的表达水平在2 cm体长时也最高,显著高于10和12 cm体长时的表达(P < 0.05)。此外,通过ISH检测到在分化性腺期间Hsd3b7主要定位在卵原细胞和精原细胞周围。这些发现首次介绍了Hsd3b7的表达特征以及Sox9a对其表达的影响,这有助于我们理解Hsd3b7在鱼类性腺中的功能。