Expression and transcriptional regulation of gsdf in spotted scat (Scatophagus argus)
Expression and transcriptional regulation of gsdf in spotted scat (Scatophagus argus)
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斑点粪便 (Scatophagus argus) 中 gsdf 的表达和转录调控
DOI：
10.1016/j.cbpb.2019.04.002
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发表时间：
2019-07-01
影响因子：
2.2
通讯作者：
Li, Guang-Li
中科院分区：
文献类型：
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作者：
Jiang, Dong-Neng;Mustapha, Umar Farouk;Li, Guang-Li
性腺体衍生因子（Gsdf）是硬骨鱼类睾丸分化和早期生殖细胞发育的关键因子。乌鳢（Scatophagus argus）具有稳定的XX-XY性别决定系统和候选性别决定基因dmrt 1，为研究硬骨鱼类性别决定和分化机制提供了良好的模型。在本研究中，我们分析了斑节对虾gsdf在性腺中的组织分布和基因表达模式，并进一步分析了转录调控。组织分布分析表明，gsdf仅在精巢和卵巢中表达。实时荧光定量PCR结果显示，gsdf和dmrt 1在不同阶段（III、IV和V期）的睾丸中的表达显著高于II、III和IV期的卵巢，而gsdf在II期卵巢中的表达显著高于III和IV期。Western blot分析也表明Gsdf在睾丸中的表达高于卵巢。免疫组化结果显示，Gsdf在睾丸精原细胞周围的支持细胞中表达，在卵巢卵原细胞周围的体细胞中表达。从斑点scat基因组DNA中克隆了gsdf的5'上游区域的约2.7kb，并且计算机启动子分析揭示了Dmrt 1和Sf 1的推定的转录因子结合位点。荧光素酶报告基因检测，使用人胚肾细胞，证明Dmrt 1激活gsdf表达的剂量依赖性的方式在Sf 1的存在下，在斑点粪便。这些结果表明，Gsdf可能参与了精原细胞和卵原细胞的发育，并可能作为Dmrt 1的下游基因参与了斑潜蝇雄性的性别分化。
Gonadal soma-derived factor (Gsdf) is critical for testicular differentiation and early germ cell development in teleosts. The spotted scat (Scatophagus argus), with a stable XX-XY sex-determination system and the candidate sex determination gene dmrt1, provides a good model for understanding the mechanism of sex determination and differentiation in teleosts. In this study, we analyzed spotted scat gsdf tissue distribution and gene expression patterns in gonads, as well as further analysis of transcriptional regulation. Tissue distribution analysis showed that gsdf was only expressed in testis and ovary. Real-time PCR showed that both gsdf and dmrt1 were expressed significantly higher in testes at different phases (phase III, IV and V) compared to ovaries at phase II, III and IV, while gsdf was expressed significantly higher in phase II ovaries than those of phase III and IV. Western blot analysis also showed that Gsdf was more highly expressed in the testis than ovary. Immunohistochemistry analysis showed that Gsdf was expressed in Sertoli cells surrounding spermatogonia in the testis, while it was expressed in the somatic cells surrounding the oogonia of the ovary. Approximately 2.7 kb of the 5' upstream region of gsdf was cloned from the spotted scat genomic DNA and in silico promoter analysis revealed the putative transcription factor binding sites of Dmrt1 and Sf1. The luciferase reporter assay, using the human embryonic kidney cells, demonstrated that Dmrt1 activated gsdf expression in a dose-dependent manner in the presence of Sf1 in spotted scat. These results suggest that Gsdf could play a role in regulating the development of spermatogonia and oogonia, and also participate in male sex differentiation by acting as a downstream gene of Dmrt1 in spotted scat.