Spata46在小鼠生精中的表达其功能过程特征及研究

发布时间：2019-06-20 19:50

【摘要】：精子发生是一个高度复杂的过程,包括有丝分裂、减数分裂和精子生成三个部分。精子生成是产生精子独有的特异性形状的重要步骤,它包括顶体形成、染色体浓缩、核塑形和鞭毛形成,最终产生高度特异的精子。精子的发育分化缺陷就能导致各种畸形精子症(如巨头精子症和圆头精子症)。精子的功能即结合卵细胞并将遗传物质传递给下一代,而畸形精子症通常与临床男性不育密切相关。基于本实验室前期的工作基础,通过睾丸组织基因表达芯片经行分析,筛选出一系列睾丸上调基因和睾丸下调基因,这些基因可能与精子发生及授精密切相关。根据人类基因组命名委员会的命名原则,我们将其中的一个基因命名为Spata46(spermatogenesis associated 46)并进行深入探讨。利用RT-PCR、RT-q PCR技术,我们发现Spata46基因在小鼠和人中均呈睾丸特异性表达,并在小鼠睾丸中呈年龄依赖性。通过免疫荧光技术,发现SPATA46特异定位于长形精子和成熟精子的头部,而细胞实验证明SPATA46为核膜蛋白。上述结果表明,SPATA46定位于精子顶体帽区域的核膜上。通过基因敲出小鼠模型,我们发现Spata46~(-/-)雄性小鼠生育力下降,并且表现为畸形精子症。进一步通过睾丸组织的透射电镜,我们发现Spata46~(-/-)精细胞的核膜断裂。最后通过体外授精实验,我们发现Spata46~(-/-)精子聚集于卵周隙,并且入卵精子数明显下降,说明SPATA46参与调控精卵融合。综上所述,SPATA46是一个新的膜蛋白,特异表达于睾丸,参与调控精细胞的核塑形及精卵融合。
[Abstract]:Spermatogenesis is a highly complex process, including mitosis, meiotic and spermatogenesis. Spermatogenesis is an important step in producing specific shapes of spermatozoa, which includes acrosome formation, chromosome concentration, nuclear shaping and flagellum formation, and finally produces highly specific spermatozoa. Defects in sperm development and differentiation can lead to a variety of abnormal spermatozoa (such as giant spermatozoa and round-headed spermatozoa). The function of spermatozoa is to bind eggs and transmit genetic material to the next generation, and abnormal spermatozoa is usually closely related to clinical male sterility. Based on the previous work in our laboratory, a series of testicular upregulation genes and testicular down-regulated genes were screened by gene expression chip analysis in testicular tissue, which may be closely related to spermatogenesis and indoctrination. According to the naming principles of the Human Genome Nomenclature Committee, one of the genes was named Spata46 (spermatogenesis associated 46 and discussed in depth. By RT-PCR,RT-q PCR, we found that Spata46 gene was specifically expressed in mice and humans, and was age-dependent in mouse testes. By immunofluorescence technique, it was found that SPATA46 was specifically located in the head of long sperm and mature sperm, and SPATA46 was proved to be a nuclear membrane protein. These results suggest that SPATA46 is located on the nuclear membrane of the sperm top cap region. By typing the mouse model, we found that the fertility of Spata46~ (- / -) male mice decreased and showed abnormal spermatozoa. Further through the transmission electron microscope of testicular tissue, we found that the nuclear membrane of Spata46~ (- / -) spermatocytes was broken. Finally, through in vitro indoctrination experiment, we found that Spata46~ (- / -) sperm gathered in the perioocyte space, and the number of sperm entering eggs decreased significantly, which indicated that SPATA46 was involved in the regulation of sperm and egg fusion. In conclusion, SPATA46 is a new membrane protein, which is specifically expressed in the testes and is involved in the regulation of spermatocyte nucleoplasmosis and sperm-egg fusion.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R698.2

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