SPATA19基因在男性生殖细胞发育过程中的表达分析及体内外研究的启示

Seyedmehdi Nourashrafeddin, Reza Ebrahimzadeh-Vesal, Mohammad Miryounesi, Mahmoud Aarabi, Nosratollah Zarghami, Mohammad Hosein Modarressi, Mohammad Nouri
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引用次数: 5

摘要

spermatogenesis -associated-19 (SPATA19)是一种新的精子发生相关基因,在生殖过程中具有重要的生物学功能。然而,SPATA19基因的表达方面尚不清楚。我们分析了SPATA19基因在小鼠睾丸体内(5、15和25日龄小鼠睾丸)发育过程中的表达,以及小鼠胚胎干细胞(ESCs)在体外(分化后5、11、19和27天)分化为雄性生殖样细胞,以及支持细胞、小鼠胚胎成纤维细胞(MEF)和NIH3T3癌细胞系中的表达。在ESC培养基中培养表达Stra8-EGFP的转基因小鼠ESCs C57BL/6J,经视黄酸(retinoic acid, RA)处理诱导其分化,然后用流式细胞仪对减数分裂细胞进行分选。RT-PCR和real-time PCR检测SPATA19 mRNA的表达。SPATA19 mRNA在体内小鼠睾丸发育过程中表达增加,在25日龄小鼠睾丸中表达最高。在RA诱导27天后,SPATA19在小鼠ESC衍生的胚样细胞和NIH3T3细胞系中表达最高,但在小鼠ESC和Sertoli细胞系中未发现表达。这些发现表明SPATA19的表达在雄性生殖细胞发育过程中增加。我们还提出了SPATA19在男性生殖细胞分化和肿瘤形成过程中的其他可能作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Analysis of SPATA19 gene expression during male germ cells development, lessons from in vivo and in vitro study

Analysis of SPATA19 gene expression during male germ cells development, lessons from in vivo and in vitro study

Spermatogenesis-associated-19 (SPATA19) is a novel spermatogenesis related gene that has important biological functions in reproduction. However, the expression aspects of SPATA19 gene are not yet well understood. We analysed SPATA19 gene expression during mouse testis development in vivo (5, 15 and 25 days old mouse testis), in parallel with mouse embryonic stem cells (ESCs) differentiation into male germ-like cells in vitro (5, 11, 19 and 27 days after differentiation), as well as Sertoli cell, mouse embryonic fibroblasts (MEF), and NIH3T3 cancerous cell line. We cultured transgenic mouse ESCs line C57BL/6J expressing Stra8-EGFP in ESC's medium and induced their differentiation by retinoic acid (RA) treatment, and meiotic cells were then sorted by FACS. RT-PCR and real-time PCR were used for analysis of SPATA19 mRNA expression. The expression of SPATA19 mRNA is increased during mouse testis development in vivo and it is highest in 25-days-old mouse testis. SPATA19 is also expressed most highly in mouse ESC-derived germ-like cells after 27 days of RA induction and NIH3T3 cell line but no expression was found in mouse ESC and Sertoli cell line. These findings demonstrate that the expression of SPATA19 increases during male germ cells development. We also suggest an additional possible role for SPATA19 in male germ cell differentiation and tumourigenic process.

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