College of Life Sciences and Engineering, Southwest University of Science and Technology, Mianyang, Sichuan 621000, China.
Department of Biosciences, COMSATS University Islamabad, Park Road, Islamabad 45550, Pakistan.
Genomics. 2024 Sep;116(5):110912. doi: 10.1016/j.ygeno.2024.110912. Epub 2024 Aug 6.
Sperm undergo a series of changes in the epididymis region before acquiring the ability to move and fertilize, and the identification of genes expressed in a region-specific manner in the epididymis provides a valuable insight into functional differences between regions. We collected epididymal tissue from three yaks and cultured epithelial cells from the caput, corpus and cauda regions of the yak epididymis using the tissue block method. RNA sequencing analysis (RNA-seq) technology was used to detect gene expression in yak epididymal caput, corpus and cauda epithelial cells. The results showed that the DEGs were highest in the caput vs. corpus comparison, and lowest in the corpus vs. cauda comparison. Six DEGs were verified by real-time fluorescence quantitative PCR (qRT-PCR), consistent with transcriptome sequencing results. The significantly enriched DNA replication pathway in the caput vs. corpus was coordinated with cell proliferation, while upregulated DEGs such as POLD1 and MCM4 were found in the DNA replication pathway. The AMPK signaling pathway was found significantly enriched in the caput vs cauda, suggesting its involvement in sperm maturation and capacitation. The TGF beta signaling pathway was screened in the corpus vs cauda and is crucial for mammalian reproductive regulation. Upregulated DEGs (TGFB3, INHBA, INHBB) are involved in the TGF beta signaling pathway. This study provides a reference for culturing yak epididymal epithelial cells in vitro, and elucidates the transcriptional profiles of epithelial cells in different segments of the epididymis, revealing the regulatory and functional differences between different segments, providing basic data for exploring the molecular mechanism of yak sperm maturation and improving the reproductive capacity of high-altitude mammals.
精子在附睾区域经历一系列变化后，才获得运动和受精的能力，而鉴定在附睾区域以特定区域方式表达的基因，为区域间的功能差异提供了有价值的见解。我们收集了三头牦牛的附睾组织，采用组织块法培养牦牛附睾头部、体部和尾部的上皮细胞。利用 RNA 测序分析（RNA-seq）技术检测了牦牛附睾头部、体部和尾部上皮细胞中的基因表达。结果表明，头部与体部比较时 DEGs 最高，体部与尾部比较时 DEGs 最低。通过实时荧光定量 PCR（qRT-PCR）验证了 6 个 DEGs，与转录组测序结果一致。头部与体部比较时显著富集的 DNA 复制途径与细胞增殖相协调，而在 DNA 复制途径中发现上调的 DEGs，如 POLD1 和 MCM4。在头部与尾部比较时，AMPK 信号通路显著富集，表明其参与精子成熟和获能。在体部与尾部比较时筛选出 TGF beta 信号通路，对于哺乳动物生殖调节至关重要。上调的 DEGs（TGFB3、INHBA、INHBB）参与了 TGF beta 信号通路。本研究为体外培养牦牛附睾上皮细胞提供了参考，阐明了附睾不同节段上皮细胞的转录谱，揭示了不同节段之间的调控和功能差异，为探索牦牛精子成熟的分子机制和提高高原哺乳动物的繁殖能力提供了基础数据。
