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小鼠中未表征的雄性生殖细胞特异性基因的表达及新型精子尾部蛋白的发现。

Expression of uncharacterized male germ cell-specific genes and discovery of novel sperm-tail proteins in mice.

作者信息

Kwon Jun Tae, Ham Sera, Jeon Suyeon, Kim Youil, Oh Seungmin, Cho Chunghee

机构信息

School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Korea.

出版信息

PLoS One. 2017 Jul 25;12(7):e0182038. doi: 10.1371/journal.pone.0182038. eCollection 2017.

Abstract

The identification and characterization of germ cell-specific genes are essential if we hope to comprehensively understand the mechanisms of spermatogenesis and fertilization. Here, we searched the mouse UniGene databases and identified 13 novel genes as being putatively testis-specific or -predominant. Our in silico and in vitro analyses revealed that the expressions of these genes are testis- and germ cell-specific, and that they are regulated in a stage-specific manner during spermatogenesis. We generated antibodies against the proteins encoded by seven of the genes to facilitate their characterization in male germ cells. Immunoblotting and immunofluorescence analyses revealed that one of these proteins was expressed only in testicular germ cells, three were expressed in both testicular germ cells and testicular sperm, and the remaining three were expressed in sperm of the testicular stages and in mature sperm from the epididymis. Further analysis of the latter three proteins showed that they were all associated with cytoskeletal structures in the sperm flagellum. Among them, MORN5, which is predicted to contain three MORN motifs, is conserved between mouse and human sperm. In conclusion, we herein identify 13 authentic genes with male germ cell-specific expression, and provide comprehensive information about these genes and their encoded products. Our finding will facilitate future investigations into the functional roles of these novel genes in spermatogenesis and sperm functions.

摘要

如果我们希望全面了解精子发生和受精的机制,那么生殖细胞特异性基因的鉴定和表征至关重要。在此,我们搜索了小鼠单基因数据库,并鉴定出13个新基因,推测它们为睾丸特异性或优势表达基因。我们的计算机模拟和体外分析表明,这些基因的表达具有睾丸和生殖细胞特异性,并且在精子发生过程中以阶段特异性方式受到调控。我们针对其中7个基因编码的蛋白质制备了抗体,以促进对雄性生殖细胞中这些蛋白质的表征。免疫印迹和免疫荧光分析表明,其中一种蛋白质仅在睾丸生殖细胞中表达,三种在睾丸生殖细胞和睾丸精子中均有表达,其余三种在睾丸阶段的精子和附睾成熟精子中表达。对后三种蛋白质的进一步分析表明,它们都与精子鞭毛中的细胞骨架结构相关。其中,预测含有三个MORN基序的MORN5在小鼠和人类精子中保守。总之,我们在此鉴定出13个具有雄性生殖细胞特异性表达的真实基因,并提供了有关这些基因及其编码产物的全面信息。我们的发现将有助于未来对这些新基因在精子发生和精子功能中的功能作用进行研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03cc/5526581/7ea98a5b4007/pone.0182038.g001.jpg

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