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MicroRNA ssa-mir-196a-4 deceases lgr8 expression in testis development of Chinese tongue sole (Cynoglossus semilaevis)

文献类型： 外文期刊

作者： Tang, Lili 1 ; You, Wuxin 3 ; Wang, Qian 1 ; Huang, Fei 5 ; Shao, Changwei 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Marine Fisheries, Minist Agr, Qingdao 266071, Peoples R China

2.Pilot Natl Lab Marine Sci & Technol Qingdao, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266237, Peoples R China

3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Single Cell Ctr CAS Key Lab Biofuels, Qingdao, Shandong, Peoples R China

4.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Key Lab Energy Genet, Qingdao, Shandong, Peoples R China

5.Genosys Inc, Shenzhen, Peoples R China

关键词： Chinese tongue sole; MicroRNA ssa-mir-196a-4; lgr8; Testis development

期刊名称：COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY （ 影响因子：2.495； 五年影响因子：2.567 ）

ISSN： 1096-4959

年卷期： 2022 年 258 卷

页码：

收录情况： SCI

摘要： MicroRNAs (miRNAs) contribute to gonadal development in animals. However, there is little information about miRNA regulation function involved in gonadal development in fish. Our group previously identified sex-related miRNAs of Chinese tongue sole (Cynoglossus semilaevis) during sex determination and differentiation by small RNA sequencing. In the present study, we characterized ssa-mir-196a-4 and its expression in testis and verified its interaction with lgr8. miRNA ssa-mir-196a-4 precursor was predicted to have a typical hairpin structure and highly conserved among various fish species. Fluorescence in situ hybridization (FISH) of ssa-mir-196a-4 in the testis of Chinese tongue sole showed that it is mainly expressed in the cytoplasm of Sertoli cells. We determined that ssa-mir-196a-4 interacted with lgr8 by bioinformatics analysis using miRanda software. According to the dual-luciferase gene reporter assay, lgr8 is a direct target of ssa-mir-196a-4. Overexpression of ssa-mir-196a-4 in the cells of the testis cell line of Chinese tongue sole decreased the expression levels of lgr8 messenger RNA (mRNA) and protein by targeting its coding sequence (CDS) region. These results suggest that ssa-mir-196a-4 acts as a post-transcriptional regulator of lgr8 and plays an important role in developing testes of Chinese tongue sole.

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