Usp9y基因在不同发育阶段小鼠睾丸组织和睾丸相关细胞系中的表达特征
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摘要
目的明确Usp9y基因在不同发育阶段小鼠睾丸组织和睾丸相关细胞系中的表达与定位并探究Usp9y的功能。方法利用生物信息学分析结合RT-PCR方法检测小鼠睾丸相关细胞系和C57BL/6J小鼠各个组织(子宫,睾丸,心,肝,脾,肺,肾,大脑,小脑,眼球)中Usp9y基因表达的特异性;利用q PCR法检测不同日龄小鼠睾丸中Usp9y基因的表达差异性。结果生物信息学分析结果显示Usp9y基因在成年小鼠睾丸中特异性表达(P <0. 05)。RT-PCR结果证实Usp9y在小鼠睾丸组织中特异性表达(P <0. 05)。usp9y在小鼠睾丸间质细胞(TM3)中特异性表达,在小鼠精原细胞(GC-1)和小鼠睾丸支持细胞(15P-1)中未检测到表达。q PCR结果显示相比其他日龄小鼠,18日龄的小鼠睾丸Usp9y表达显著升高(P <0. 05)。结论通过RT-PCR的方法确定Usp9y基因在TM3细胞中特异性表达;小鼠Usp9y基因在小鼠出生后18 d表达明显增高,上述结果表明Usp9y基因在精子发生早期发挥着不可替代的作用。
Objective To clarify the expression and localization of Usp9 y gene in mouse testis and testicular-related cell lines at different developmental stages and to explore the function of Usp9 y. Methods Bioinformatics analysis combined with RT-PCR was used to detect Usp9 y gene expression in mouse testis-associated cell lines and C57 BL/6 J mouse tissues( uterus,testis,heart,liver,spleen,lung,kidney,brain,cerebellum,eyeball). The expression of Usp9 y gene in testes of mice at different developmental stages was detected by q PCR method. Results Bioinformatics analysis showed that Usp9 y was specifically expressed in the testis of the adult mouse( P < 0. 05). RT-PCR results confirmed the specific expression. Usp9 y was specifically expressed in mouse testicular leydig cell( TM3),while it was not detected in spermatogonia( GC-1) and Sertoli cells( 15 P-1). The q PCR results showed that the expression of Usp9 y in the testis of mice at day 18 was significantly increased( P < 0. 05). Conclusion The specific expression of Usp9 y gene can be detected in mouse testicular Leydig cells by RT-PCR method. The expression of Usp9 y gene is significantly higher at the postnatal day 18,indicating that Usp9 y gene may play a significant role in the early stage of spermatogenesis.
Objective To clarify the expression and localization of Usp9 y gene in mouse testis and testicular-related cell lines at different developmental stages and to explore the function of Usp9 y. Methods Bioinformatics analysis combined with RT-PCR was used to detect Usp9 y gene expression in mouse testis-associated cell lines and C57 BL/6 J mouse tissues( uterus,testis,heart,liver,spleen,lung,kidney,brain,cerebellum,eyeball). The expression of Usp9 y gene in testes of mice at different developmental stages was detected by q PCR method. Results Bioinformatics analysis showed that Usp9 y was specifically expressed in the testis of the adult mouse( P < 0. 05). RT-PCR results confirmed the specific expression. Usp9 y was specifically expressed in mouse testicular leydig cell( TM3),while it was not detected in spermatogonia( GC-1) and Sertoli cells( 15 P-1). The q PCR results showed that the expression of Usp9 y in the testis of mice at day 18 was significantly increased( P < 0. 05). Conclusion The specific expression of Usp9 y gene can be detected in mouse testicular Leydig cells by RT-PCR method. The expression of Usp9 y gene is significantly higher at the postnatal day 18,indicating that Usp9 y gene may play a significant role in the early stage of spermatogenesis.
引文
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[12] Bhasin S,De Kretser DM,Baker HW. Clinical review 64:pathology and natural history of male infertility[J]. J Clin Endocrind Metab,1994,79(6):1525-1529.
[13] Jamsai D,O’Bryan MK. Mouse models in male fertility research[J]. Asian J Androl,2011,13(1):139-151.
[14] Nakasuji T,Ogonuki N,Chiba T,et al. Complementary critical functions of Zfy1 and Zfy2 in mouse spermatogenesis and reproduction[J]. PLo S Genetics,2017,13(1):e1006578.
[15]袁双虎,徐斯凡.睾丸间质细胞凋亡及调控[J].中华男科学,2003,9(3):218-221.
[16] Kaur G,Vadala S,Dufour JM. An overview of a Sertoli cell transplantation model to study testis morphogenesis and the role of the Sertoli cells in immune privilege[J]. Envir Epigenet,2017,3(3):1-10.
[2]张香改,黄海燕,刘杰等.男性不育与Y染色体微缺失关系的研究进展[J].中国计划生育学杂志,2007,15(9):570-572.
[3]高迎春,张传领,肖瑞. USP9Y基因在男性不育等疾病中的研究进展[J].生殖与避孕,2015,35(5):333-338.
[4] Brown GM,Furlong RA,Sargent CA,et al. Characterisation of the coding sequence and fine mapping of the human DFFRY gene and comparative expression analysis and mapping to the Sxrb interval of the mouse Y chromosome of the Dffry gene[J]. Hum Mol Genet,1998,7(1):97-107.
[5]张丽芳,曾艳. AZF基因微缺失与男性不育研究进展[J].中国优生与遗传杂志,2012,20(3):4-6.
[6] Sun C,Skaletsky H,Birren B,et al. An azoospermic man with a de novo point mu-tation in the Y-chromosomal gene USP9Y[J].Nat Genet,1999,23(4):429-432.
[7] Roth G,Himstedt W. Response characteristics of neurons in the tectum opticum of Salamandra[J]. Naturwissenschaften,1978,65(12),657-658.
[8] Hou CC,Yang WX. New insights to the ubiquitin-proteasome pathway(UPP)mechanism during spermatogenesis[J]. Mol Biol Rep,2013,40(4),3213-3230.
[9]王英男.精子功能检测对选择IVF或ICSI治疗不育症的临床意义[J].现代妇女:医学前沿,2015,(2):112.
[10]陈瑞,于帅,陈晓旭,等.非编码RNA对哺乳动物精子发生过程的调控[J].中国农业科学,2017,50(2):380-390.
[11]吴际.精原细胞向生殖干细胞和生殖细胞转化的机制研究[J].中国基础科学,2017,19(2):10-17.
[12] Bhasin S,De Kretser DM,Baker HW. Clinical review 64:pathology and natural history of male infertility[J]. J Clin Endocrind Metab,1994,79(6):1525-1529.
[13] Jamsai D,O’Bryan MK. Mouse models in male fertility research[J]. Asian J Androl,2011,13(1):139-151.
[14] Nakasuji T,Ogonuki N,Chiba T,et al. Complementary critical functions of Zfy1 and Zfy2 in mouse spermatogenesis and reproduction[J]. PLo S Genetics,2017,13(1):e1006578.
[15]袁双虎,徐斯凡.睾丸间质细胞凋亡及调控[J].中华男科学,2003,9(3):218-221.
[16] Kaur G,Vadala S,Dufour JM. An overview of a Sertoli cell transplantation model to study testis morphogenesis and the role of the Sertoli cells in immune privilege[J]. Envir Epigenet,2017,3(3):1-10.
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