State Key Laboratory of Molecular Biology, Shanghai Key Laboratory of Molecular Andrology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China.
Beijing Advanced Innovation Center for Genomics, Biomedical Institute for Pioneering Investigation via Convergence, College of Life Sciences, Peking University, Beijing, 100871, China.
Cell Res. 2018 Sep;28(9):879-896. doi: 10.1038/s41422-018-0074-y. Epub 2018 Jul 30.
A systematic interrogation of male germ cells is key to complete understanding of molecular mechanisms governing spermatogenesis and the development of new strategies for infertility therapies and male contraception. Here we develop an approach to purify all types of homogeneous spermatogenic cells by combining transgenic labeling and synchronization of the cycle of the seminiferous epithelium, and subsequent single-cell RNA-sequencing. We reveal extensive and previously uncharacterized dynamic processes and molecular signatures in gene expression, as well as specific patterns of alternative splicing, and novel regulators for specific stages of male germ cell development. Our transcriptomics analyses led us to discover discriminative markers for isolating round spermatids at specific stages, and different embryo developmental potentials between early and late stage spermatids, providing evidence that maturation of round spermatids impacts on embryo development. This work provides valuable insights into mammalian spermatogenesis, and a comprehensive resource for future studies towards the complete elucidation of gametogenesis.
系统研究雄性生殖细胞对于全面了解精子发生的分子机制以及开发新的不孕治疗和男性避孕策略至关重要。在这里，我们开发了一种通过结合转基因标记和生精上皮周期同步化，以及随后的单细胞 RNA 测序来纯化所有类型同质精原细胞的方法。我们揭示了广泛的、以前未被描述的基因表达动态过程和分子特征，以及特定的选择性剪接模式，以及雄性生殖细胞发育特定阶段的新调控因子。我们的转录组学分析使我们能够发现用于在特定阶段分离圆形精子细胞的有区别的标记物，以及早期和晚期精子细胞之间不同的胚胎发育潜力，这为精子细胞成熟对胚胎发育的影响提供了证据。这项工作为哺乳动物精子发生提供了有价值的见解，并为未来全面阐明配子发生的研究提供了全面的资源。
