Table_5_Transcriptome Profiling of the Ovarian Cells at the Single-Cell Resolution in Adult Asian Seabass.XLSX

Single-cell RNA sequencing (scRNA-seq) is widely adopted for identifying the signature molecular markers or regulators in cells, as this would benefit defining or isolating various types of cells. Likewise, the signature transcriptome profile analysis at the single cell level would well illustrate the key regulators or networks involved in gametogenesis and gonad development in animals; however, there is limited scRNA-seq analysis on gonadal cells in lower vertebrates, especially in the sexual reversal fish species. In this study, we analyzed the molecular signature of several distinct cell populations of Asian seabass adult ovaries through scRNA-seq. We identified five cell types and also successfully validated some specific genes of germ cells and granulosa cells. Likewise, we found some key pathways involved in ovarian development that may concert germline-somatic interactions. Moreover, we compared the transcriptomic profiles across fruit fly, mammals, and fish, and thus uncovered the conservation and divergence in molecular mechanisms that might drive ovarian development. Our results provide a basis for studying the crucial features of germ cells and somatic cells, which will benefit the understandings of the molecular mechanisms behind gametogenesis and gonad development in fish.

单细胞RNA测序（single-cell RNA sequencing，scRNA-seq）已被广泛应用于鉴定细胞内的标志性分子标志物与调控因子，这将助力各类细胞的定义与分离。同样地，单细胞水平的特征转录组谱分析，可清晰阐明动物配子发生与性腺发育过程中涉及的关键调控因子及调控网络。然而目前针对低等脊椎动物性腺细胞的scRNA-seq研究仍较为匮乏，尤其在性反转鱼类中相关报道极少。本研究通过scRNA-seq技术，分析了成年亚洲海鲈卵巢内多个不同细胞群的分子特征，共鉴定出5种细胞类型，并成功验证了生殖细胞与颗粒细胞的部分特异性基因。此外，本研究还发现了若干参与卵巢发育的关键通路，这些通路可能协同调控生殖细胞与体细胞间的相互作用。进一步地，本研究比对了果蝇、哺乳动物与鱼类的转录组谱，由此揭示了驱动卵巢发育的分子机制的保守性与差异性。本研究结果为解析生殖细胞与体细胞的关键特征提供了重要研究基础，将有助于深入理解鱼类配子发生与性腺发育背后的分子机制。