Yao Jingting, Li Sendong, Ban Wenzhuo, Zeng Linhui, Cui Han, Chen Kaili, Xu Hongyan
Integrative Science Center of Germplasm Creation in Western China Integrative Science Center of Germplasm Creation in Western China (CHONGQING) Science City & College of Fisheries, Southwest University, Key Laboratory of Freshwater Fish Reproduction and Development, Ministry of Education, Key Laboratory of Aquatic Sciences of Chongqing, Chongqing, 402460, China.
Mar Biotechnol (NY). 2025 Jun 23;27(4):101. doi: 10.1007/s10126-025-10474-5.
Spermatogenesis is a complicated process of sexual reproduction, involving cell proliferation and differentiation, and depends on intricate interactions between testicular somatic cells and germ cells. For further investigations on male germ cells' development and differentiation in Chinese soft-shelled turtle (Pelodiscus sinensis), it is crucial to define testicular cell types and their molecular regulators. Here, the 10 × Genomics single-cell RNA sequencing was adopted to study the transcriptomic profiles of single cells during the spermatogenesis of adult Chinese soft-shelled turtle. In total, 7317 individual cell transcriptomes were collected for analysis, and 11 cell types were identified with known differentially expressed genes, including Leydig cells, Sertoli cells, spermatogonia, spermatocytes, and spermatids. Likewise, the top 10 marker genes and top-enriched gene pathways were analyzed in each cell type. Intriguingly, ligand-receptor analysis showed that the strongest interaction between Leydig cells and germ cells was using CellChat. Moreover, a primary developmental trajectory of male germ cells was constructed from spermatogonia to spermatids, as well as some important cell-specific regulators were identified for labeling the germ cells at different stages, including PCNA and Stra8, validated by immunostaining fluorescence. In addition, the transcriptomic profiles of male germ cells at different stages were comparatively analyzed among fruit fly, fish, mammals, and Chinese soft-shelled turtle, and the conserved and divergent regulators of male germ cells were summarized across species. In conclusion, this study provided novel insights into the testicular cells' atlas in turtles, and the findings would facilitate the development of techniques for manipulating germ cells, such as isolating cells and defining stages of differentiation.
精子发生是一个复杂的有性生殖过程，涉及细胞增殖和分化，并且依赖于睾丸体细胞与生殖细胞之间复杂的相互作用。为了进一步研究中华鳖雄性生殖细胞的发育和分化，定义睾丸细胞类型及其分子调节因子至关重要。在此，采用10×基因组学单细胞RNA测序技术研究成年中华鳖精子发生过程中单个细胞的转录组图谱。总共收集了7317个单细胞转录组进行分析，通过已知的差异表达基因鉴定出11种细胞类型，包括睾丸间质细胞、支持细胞、精原细胞、精母细胞和精子细胞。同样，对每种细胞类型中的前10个标记基因和富集程度最高的基因通路进行了分析。有趣的是，配体-受体分析表明，使用CellChat软件发现睾丸间质细胞与生殖细胞之间的相互作用最强。此外，构建了从精原细胞到精子细胞的雄性生殖细胞的主要发育轨迹，并且鉴定出一些重要的细胞特异性调节因子用于标记不同阶段的生殖细胞，包括增殖细胞核抗原（PCNA）和视黄酸诱导基因8（Stra8），通过免疫荧光染色进行了验证。此外，对果蝇、鱼类、哺乳动物和中华鳖不同阶段雄性生殖细胞的转录组图谱进行了比较分析，总结了跨物种的雄性生殖细胞保守和不同的调节因子。总之，本研究为龟类睾丸细胞图谱提供了新的见解，这些发现将有助于开发操纵生殖细胞的技术，如分离细胞和定义分化阶段。
