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A Study of Differential Expression of Testicular Genes in Various Reproductive Phases of Hemidactylus flaviviridis (Wall Lizard) to Derive Their Association with Onset of Spermatogenesis and Its Relevance to Mammals

机译:黄皮半球不同生殖期睾丸基因差异表达的研究及其与精子发生的关系及其与哺乳动物的关系

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摘要

Testis of Hemidactylus flaviviridis, commonly known as Indian wall lizard, displays a lack of cellular and metabolic activity in regressed phase of testis during non-breeding season of the year. Retracted Sertoli cells (Sc), fibroid myoid cells and pre-meiotic resting spermatogonia are observed in such testis. This situation is akin to certain forms of infertility in men where hormone supplementation fails to generate sperm despite the presence of Sc and germ cells (Gc) in testis. In testis of lizard, spermatogenesis is reinitiated upon increased level of hormones during appropriate season (phase of recrudescence). Study of genes associated with generation of sperm, from regressed adult testis in lizard, may provide valuable information for understanding certain forms of male idiopathic infertility. Subtractive hybridization using testicular RNA obtained from the regressed and active phases of lizard reproductive cycle led to identify eight partial mRNA sequences that showed sequence homology with mice genes. We further evaluated the gene expression prolife by real-time PCR in three different reproductive phases of H. flaviviridis: regressed (pre-meiotic), recrudescent (meiotic) and active (post meiotic), for comparison with the corresponding testicular phases found in testis of 5 days (pre-meiotic), 20 days (meiotic) and 60 days (post-meiotic) old mouse. This is the first report where genes associated with progression of spermatogenesis during active phase, which follows a regressed state of adult testis, were identified in lizard and found to be conserved in mouse. Six important genes, Hk1, Nme5, Akap4, Arih1, Rassf7 and Tubb4b were found to be strictly associated with active spermatogenesis in both mouse and lizard. Factors interfering with the expression of any of these genes may potentially abrogate the process of spermatogenesis leading to infertility. Such information may shed light on unknown causes of idiopathic male infertility.
机译:每年在非繁殖季节,黄皮半球睾丸(通常被称为印度壁蜥)在睾丸的退缩期显示缺乏细胞和代谢活性。在这种睾丸中观察到收缩的支持细胞(Sc),肌瘤样肌细胞和减数分裂前的静息精原细胞。这种情况类似于男性不育的某些形式,尽管睾丸中存在Sc和生殖细胞(Gc),但激素补充不能产生精子。在蜥蜴的睾丸中,在适当的季节(再发阶段),荷尔蒙水平升高,精子发生重新开始。蜥蜴退化成年睾丸中与精子生成相关的基因研究可能为理解某些形式的男性特发性不育症提供有价值的信息。使用从蜥蜴生殖周期的退化期和活跃期获得的睾丸RNA进行减性杂交,可以鉴定出八个部分mRNA序列,这些序列与小鼠基因具有同源性。我们进一步通过实时PCR评估了黄褐线虫的三个不同生殖阶段(退化(减数分裂前),衰退(减数分裂)和活跃(减数分裂后))的基因表达增殖,以与睾丸中相应的睾丸期进行比较。 5天(减数分裂前),20天(减数分裂)和60天(减数分裂后)的老鼠。这是第一份报告,其中在蜥蜴中发现了与活跃期精子发生进展相关的基因,该基因遵循成年睾丸的退化状态,并在小鼠中被保存。六个重要的基因,Hk1,Nme5,Akap4,Arih1,Rassf7和Tubb4b被发现与小鼠和蜥蜴中活跃的精子发生密切相关。干扰任何这些基因表达的因素都可能会废除导致不育的精子发生过程。这些信息可能会揭示特发性男性不育的未知原因。

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