PDF(Pubmed)
Abstract：
The development and maturation of sperm entails intricate metabolic processes involving water molecules, amino acids, hormones, and various substances. Among these processes, the role of aquaporins (aqps) in the testis is crucial. Turbot (Scophthalmus maximus) is a significant marine flatfish species in China; however, natural egg laying in females is not feasible under cultured conditions. Consequently, artificial insemination becomes necessary, requiring the retrieval of sperm and eggs through artificial methods. In this study, we combined genomic, transcriptomics, RT-qPCR, computer-assisted sperm analysis (CASA), and immunohistochemistry to investigate the involvement of the aqp family in spermatogenesis in turbot. Through genomic data analysis, we identified 16 aqps genes dispersed across 13 chromosomes, each exhibiting the characteristic major intrinsic protein (MIP) domain associated with AQPs. The results from RNA-seq and RT-qPCR analysis revealed prominent expression of aqp4, 10, and 12 during the proliferative stage, whereas aqp1 showed primary expression during the mature stage. aqp11 displayed high expression levels during both MSII and MSV stages, potentially contributing significantly to the proliferation and maturation of male germ cells. Conversely, aqp8 showed elevated expression levels during the MSIII, MSIII-IV, and MSIV stages, suggesting its direct involvement in spermiogenesis. Immunohistochemical analysis unveiled the predominant localization of AQP1 protein in male germ cells rather than Sertoli cells, specifically concentrated in the head of sperm within cysts. Furthermore, a noteworthy decline in sperm motility was observed when sperm were subjected to treatment with either the AQP1-specific inhibitor (HgCl2) or the AQP1 antibody. However, no direct correlation was found between the expression of Smaqp1 and sperm quality. Overall, these findings provide new insights into the involvement of aqps in teleost spermatogenesis. Moreover, they hold potential for improving techniques related to sperm activation and cryopreservation, offering valuable knowledge for future advancements in this field.
摘要：
精子的发育和成熟需要复杂的代谢过程，涉及水分子，氨基酸,荷尔蒙，和各种物质。在这些过程中，水通道蛋白（aqps）在睾丸中的作用至关重要。Turbot(Scophthalmusmaximus)是中国重要的海洋比目鱼；然而，在养殖条件下，雌性自然产卵是不可行的。因此,人工授精变得必要，需要通过人工方法获取精子和卵子。在这项研究中,我们结合了基因组，转录组学，RT-qPCR,计算机辅助精子分析（CASA），和免疫组织化学研究aqp家族在大菱头精子发生中的参与。通过基因组数据分析,我们确定了分散在13条染色体上的16个aqps基因，每个都表现出与AQP相关的特征性主要内在蛋白(MIP)结构域。来自RNA-seq和RT-qPCR分析的结果显示aqp4、10和12在增殖阶段的显著表达,而aqp1在成熟期显示出初级表达。aqp11在MSII和MSV阶段都显示出高表达水平，可能对雄性生殖细胞的增殖和成熟做出重大贡献。相反，aqp8在MSIII期间显示表达水平升高，MSIII-IV,和MSIV阶段，表明它直接参与精子发生。免疫组织化学分析揭示了AQP1蛋白在雄性生殖细胞而不是支持细胞中的主要定位。特别集中在头部的精子内的囊肿。此外，当精子接受AQP1特异性抑制剂(HgCl2)或AQP1抗体治疗时,观察到精子活力显著下降.然而,Smaqp1的表达与精子质量无直接相关性。总的来说,这些发现为aqps参与硬骨鱼精子发生提供了新的见解。此外，它们有可能改进与精子激活和冷冻保存相关的技术，为该领域的未来发展提供有价值的知识。
