摘要
目的:研究去泛素化酶24(USP24)基因在小鼠睾丸精子发生过程中的表达特征,初步探讨其在生精过程中的作用。方法:通过实时荧光定量PCR(qPCR)和免疫荧光等方法检测USP24在不同周龄野生型小鼠睾丸组织以及成年雄激素受体敲除(ARKO)小鼠睾丸中的表达特征;采用双荧光素酶报告基因实验检测USP24启动子转录活性。结果:qPCR和免疫荧光结果表明,USP24基因在出生1周时表达水平较低,3周时急剧升高,随后维持在相似水平至第8周;USP24主要定位于支持细胞和生精细胞的细胞质;与野生型相比,USP24在ARKO小鼠睾丸表达降低;性成熟小鼠睾丸中,USP24定位于成熟精子头部后端及中部。双荧光素酶报告基因实验结果显示,睾酮刺激后USP24启动子的转录活性升高。结论:USP24基因的表达水平的升高与小鼠的性发育相关,且其蛋白在小鼠成熟精子上表达。USP24是受雄激素受体(AR)调控的靶基因。USP24可能参与调控小鼠的精子发生过程。
Objective: To investigate the expression characteristics of the USP24 gene in the mouse testis and its role in sper- matogenesis. Methods : We examined the expression characteristics of USP24 in the testis tissues of wild-type mice at different post- natal weeks (PNW) and androgen receptor (AR)-knockout (ARKO) adult mice using real-time quantitative PCR and immunofluores- cence, and detected the transcriptional activity of the USP24 promoter by dual-luciferase reporter gene assay. Results: The expres- sion of the USP24 gene was low in the testis tissue of the wild-type mice at PNW 1, increased dramatically at PNW 3 and stayed at a similar level till PNW 8. The USP24 protein was located mainly in the cytoplasm of Sertoli and spermatogenic cells. Compared with the wild-type, the adult ARKO mice showed a decreased expression of USP24 localized in the posterior head and mid-piece of the mature sperm in the testis. Dual-luciferase reporter gene assay showed that the transcriptional activity of the USP24 promoter was increased af- ter testosterone stimulation. Conclusion : The increased expression of the USP24 gene was associated with the initiation of sexual de- velopment, and the USP24 protein was expressed in the mature sperm of the mice. USP24 is an AR-target gene, which may be in- volved in the regulation of spermatogenesis in mice.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2017年第11期963-968,共6页
National Journal of Andrology
基金
深圳市知识创新计划项目(JCYJ20150402144905865)~~