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在小鼠睾丸发育减数分裂期之后表达的Spata-19新变体的鉴定。

Identification of Spata-19 new variant with expression beyond meiotic phase of mouse testis development.

作者信息

Nourashrafeddin Seyedmehdi, Ebrahimzadeh-Vesal Reza, Modarressi Mohammad Hosein, Zekri Ali, Nouri Mohammad

机构信息

Department of Clinical Biochemistry, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Rep Biochem Mol Biol. 2014 Apr;2(2):89-93.

Abstract

BACKGROUND

The study of specific genes expressed in the testis is important to understanding testis development and function. Spermatogenesis is an attractive model for the study of gene expression during germ cell differentiation. Spermatogenesis associated-19 (Spata-19) is a recently-identified important spermatogenesis-related gene specifically expressed in testis. Its protein product is involved in sperm cell development and reproduction. In this report we examined the expression of Spata-19 mRNA in mouse testis, fetus, and cell lines.

METHODS

Reverse transcription-polymerase chain reaction (RT-PCR), nested PCR, and PCR-restriction fragment length polymorphism (PCR-RFLP) were used to analyze Spata-19 mRNA expression in different stages of mouse testis development, mouse fetus, mouse embryonic fibroblasts (MEF), mouse embryonic stem cells (mESC), Sertoli cells, and NIH/3T3 cells.

RESULTS

We identified a novel splice variant of Spata-19 in the mouse genome that it is expressed in the fetus and after the meiotic phase of spermatogenesis, and over-expressed in the post-meiotic stage of mouse spermatogenesis. This novel splice variant was absent in five days old mice testis, mESC, MEF, Sertoli, and NIH/3T3 cell lines.

CONCLUSION

The Spata-19 has a large novel splice variant in mouse testis that is expressed beyond meiotic phase of testis development. We suggest that this new Spata-19 mRNA variant might be involved in mitochondrial maintenance in sperm cells, and might be correlated with androgen secretion ‎and male fertility.

摘要

背景

研究睾丸中特异性表达的基因对于理解睾丸发育和功能很重要。精子发生是研究生殖细胞分化过程中基因表达的一个有吸引力的模型。精子发生相关-19(Spata-19)是最近鉴定出的一个在睾丸中特异性表达的重要精子发生相关基因。其蛋白质产物参与精子细胞的发育和繁殖。在本报告中,我们检测了Spata-19 mRNA在小鼠睾丸、胎儿及细胞系中的表达情况。

方法

采用逆转录-聚合酶链反应(RT-PCR)、巢式PCR及PCR-限制性片段长度多态性(PCR-RFLP)分析Spata-19 mRNA在小鼠睾丸发育不同阶段、小鼠胎儿、小鼠胚胎成纤维细胞(MEF)、小鼠胚胎干细胞(mESC)、支持细胞及NIH/3T3细胞中的表达。

结果

我们在小鼠基因组中鉴定出Spata-19的一个新的剪接变体,它在胎儿及精子发生减数分裂期之后表达,并在小鼠精子发生的减数分裂后阶段过表达。这个新的剪接变体在5日龄小鼠睾丸、mESC、MEF、支持细胞及NIH/3T3细胞系中不存在。

结论

Spata-19在小鼠睾丸中有一个大的新剪接变体,其在睾丸发育的减数分裂期之后表达。我们认为这个新的Spata-19 mRNA变体可能参与精子细胞中的线粒体维持,并且可能与雄激素分泌及男性生育力相关。

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