Transcription factors foxl2 and foxl3 regulate cyp19a1a and cyp11b in orange-spotted grouper (Epinephelus coioides)
Transcription factors foxl2 and foxl3 regulate cyp19a1a and cyp11b in orange-spotted grouper (Epinephelus coioides)
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DOI：
10.1016/j.aqrep.2022.101243
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发表时间：
2022-08
影响因子：
3.7
通讯作者：
Chunli Zhang;Qi He;Huitao Cheng;Lihua Li;Xinhe Ruan;Xuzhuo Duan;Fengqi Huang;Huirong Yang;Haifa Zhang;Herong Shi;Qing Wang;Huihong Zhao
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文献类型：
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作者：
Chunli Zhang;Qi He;Huitao Cheng;Lihua Li;Xinhe Ruan;Xuzhuo Duan;Fengqi Huang;Huirong Yang;Haifa Zhang;Herong Shi;Qing Wang;Huihong Zhao
在橙点石斑鱼(E.coioides)中，转录因子fox12和fox13被认为在卵巢和睾丸发育中发挥重要作用，但它们是如何发挥作用的尚不清楚。在本研究中，我们研究了ox12和fox13对性类固醇基因yp19a1a和cyp11b的调节作用，并探讨了ox12和fox13在硬骨鱼性激素合成途径中的作用。首先，我们利用FOXL2和FOX13重组蛋白研究了FOXL2/FOX13对芳香酸酶的调控机制。并发现重组蛋白可以进入性腺细胞并在细胞核中表达。用FOXL2或FOXL3重组蛋白刺激原代卵巢细胞，用实时荧光定量聚合酶链式反应检测cyp19a1a和cyp11b的表达水平。FOXL2重组蛋白显著增加与雌激素产生相关的cyp19a1amRNA的表达，并呈剂量依赖关系。Fox13重组蛋白显著下调cyp19a1amRNA的表达，显著上调与雄激素合成相关的cyp11bmRNA的表达。体外双荧光素酶分析表明FOXL2直接作用于cyp19a1a的启动子区域，foxl3直接作用于cyp19a1a和cyp11b的启动子区域。体内注射FOXL2和FOXL3重组蛋白的手术实验进一步表明，FOXL2通过直接上调芳香酶基因cyp19a1a的表达促进卵巢分化，而foxl3直接抑制芳香酶基因cyp19a1a的表达和上调cyp11b的表达促进睾丸分化。
In orange-spotted grouper (E. coioides),the transcription factorsfoxl2andfoxl3are thought to play an important role in ovarian and testicular development, but how they do so remains unclear. In this study, we investigated the regulatory effect offoxl2andfoxl3on the sex steroid genescyp19a1aandcyp11b, and then we explored the function offoxl2andfoxl3in the sex hormone synthesis pathway in teleost fish. First, we used FOXL2 and FOXL3 recombinant proteins to investigate the regulatory mechanism offoxl2/foxl3on aromatase inE. coioidesand found that the recombinant proteins can enter gonad cells and be expressed in the nucleus. The expression levels ofcyp19a1aandcyp11bwere detected by real-time fluorescent quantitative PCR when FOXL2 or FOXL3 recombinant proteins were used to stimulate primary ovarian cells of orange-spotted grouper. The FOXL2 recombinant protein significantly increased the expression ofcyp19a1amRNA, which is related to estrogen production, in a dose-dependent manner. The FOXL3 recombinant protein significantly down-regulated the expression ofcyp19a1amRNA and significantly up-regulated the expression ofcyp11bmRNA, which is related to androgen synthesis. Dual luciferase analysis in vitro revealed that foxl2 directly acted on the promoter region of cyp19a1a and foxl3 directly acted on the promoter regions of both cyp19a1a and cyp11b. The FOXL2 and FOXL3 recombinant protein injection experiments in vivo by surgery further showed that foxl2 promoted ovarian differentiation by directly up-regulating the expression of the aromatase gene cyp19a1a, whereas foxl3 directly inhibited the expression of the aromatase gene cyp19a1a and up-regulated the expression of cyp11b to promote testicular differentiation.