miR-9-5p 调节参与小鼠睾丸发育和精子发生的 Sirt1

IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY Theriogenology Pub Date : 2024-09-10 DOI:10.1016/j.theriogenology.2024.09.007
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引用次数: 0

摘要

睾丸发育和精子生成对男性生殖至关重要,而组蛋白(脱)乙酰化在生殖细胞内的染色质重塑中起着关键作用。Sirt1是一种关键的组蛋白去乙酰化酶,与染色质重塑有关,但它在睾丸发育和精子发生中的表达模式和具体作用还需要进一步研究。本研究利用 RT-qPCR、Western 印迹、免疫荧光和细胞转染等方法,全面分析了 Sirt1 在成年和幼年小鼠睾丸组织以及各种雄性生殖细胞中的表达。研究人员进行了 GO 和 KEGG 富集分析,以阐明与 Sirt1 及其相关基因有关的生物学功能和通路。利用多个 miRNA 数据库预测了靶向 Sirt1 的 miRNA,并通过 RT-qPCR 验证了它们的表达水平。慢病毒转染用于敲除候选的 miRNA,以评估它们的功能作用。结果发现,与幼鼠组织相比,Sirt1在成年小鼠睾丸组织中的表达明显下调,而且在不同的雄性生殖细胞中存在明显差异。Sirt1在精原细胞和成熟精子中的表达量很高,但在精母细胞和精子细胞中的表达量相对较低。GO和KEGG富集分析强调了Sirt1在染色质组织、细胞增殖调控和能量平衡等关键生物过程中的作用,以及它与细胞衰老、FoxO信号通路和AMPK信号通路等信号通路的关联。生物信息学分析和随后的 RT-qPCR 验证确定了 miR-9-5p 是一种靶向 Sirt1 的 miRNA。在成年小鼠睾丸组织中,miR-9-5p的表达明显高于幼年组织,与Sirt1的水平成反比。此外,敲除 miR-9-5p 会导致 Sirt1 mRNA 和蛋白质表达的显著增加。总之,Sirt1 是小鼠睾丸发育和精子生成过程中的关键角色。miR-9-5p 负向调控 Sirt1 的发现表明,这些过程可能受一个关键调控轴的支配,从而为男性生育力提供了新的见解,并为治疗干预提供了潜在靶点。
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miR-9-5p regulates Sirt1 involved in testicular development and spermatogenesis in mouse

Testicular development and spermatogenesis are critical for male reproduction, with histone (de)acetylation playing a key role in chromatin remodeling within germ cells. Sirt1, a key histone deacetylase, is implicated in chromatin remodeling, but its expression pattern and specific role in testicular development and spermatogenesis need further study. This study comprehensively analyzed Sirt1 expression in adult and juvenile mouse testicular tissues and across various male germ cells, utilizing RT-qPCR, Western blot, immunofluorescence, and cell transfection. GO and KEGG enrichment analyses were performed to elucidate the biological functions and pathways associated with Sirt1 and its related genes. Multiple miRNA databases were utilized to predict miRNAs targeting Sirt1, and their expression levels were validated using RT-qPCR. Lentiviral transfection was used to knockdown candidate miRNAs to assess their functional roles. The results revealed a significant downregulation of Sirt1 expression in adult mouse testicular tissues compared to juvenile tissues, with pronounced variation across diverse male germ cells. Sirt1 was highly expressed in spermatogonia and mature sperm, but comparatively lower in spermatocytes and spermatids. GO and KEGG enrichment analyses highlighted Sirt1's role in key biological processes, including chromatin organization, regulation of cell proliferation, and energy homeostasis, as well as its association with signaling pathways like cellular senescence, the FoxO signaling pathway, and the AMPK signaling pathway. Bioinformatic analysis and subsequent RT-qPCR validation identified miR-9-5p as a miRNA targeting Sirt1. The expression of miR-9-5p was significantly higher in adult mouse testicular tissues compared to juvenile tissues, inversely correlating with Sirt1 levels. Moreover, the knockdown of miR-9-5p led to a notable increase in Sirt1 mRNA and protein expression. In conclusion, Sirt1 is a key player in mouse testicular development and spermatogenesis. The discovery that miR-9-5p negatively regulates Sirt1 suggests a critical regulatory axis that may govern these processes, providing novel insights into male fertility and potential targets for therapeutic intervention.

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来源期刊
Theriogenology
Theriogenology 农林科学-生殖生物学
CiteScore
5.50
自引率
14.30%
发文量
387
审稿时长
72 days
期刊介绍: Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.
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