Single-cell transcriptome reveals insights into the development and function of the zebrafish ovary.
Single-cell transcriptome reveals insights into the development and function of the zebrafish ovary.
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DOI：
10.7554/elife.76014
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发表时间：
2022-05-19
期刊：
影响因子：
7.7
通讯作者：
Postlethwait, John
中科院分区：
文献类型：
--
作者：
Liu, Yulong;Kossack, Michelle E.;McFaul, Matthew E.;Christensen, Lana N.;Siebert, Stefan;Wyatt, Sydney R.;Kamei, Caramai N.;Horst, Samuel;Arroyo, Nayeli;Drummond, Iain A.;Juliano, Celina E.;Draper, Bruce W.;Postlethwait, John
关键词：
斑马鱼是一种成熟的研究生物，为我们理解脊椎动物组织和器官发育做出了许多贡献，但我们对调节性腺发育，性别和生殖的基因的理解仍然存在重大差距。与许多器官的发育不同，例如在发育的最初几天形成的大脑和心脏，斑马鱼性腺直到幼虫阶段（受精后≥5天）才开始形成。因此，正向遗传筛选已经确定了非常少的性腺发育所需的基因。此外，批量RNA测序研究，确定基因表达的性腺没有必要的决议，以确定少数细胞群体，可能发挥重要作用，这些器官的发展和功能。为了克服这些限制，我们使用单细胞RNA测序来确定从幼年斑马鱼卵巢分离的细胞的转录组。这导致了10，658个生殖细胞和14，431个体细胞的概况。我们的生殖细胞数据代表了从生殖干细胞到早期减数分裂卵母细胞的所有发育阶段。我们的体细胞数据代表了所有已知的体细胞类型，包括卵泡细胞、卵泡膜细胞和卵巢基质细胞。进一步的分析显示，在这些广泛定义的细胞类型中，细胞亚群的数量出乎意料。为了进一步确定它们的功能意义，我们确定了这些细胞亚群在卵巢中的位置。最后，我们使用基因敲除实验来确定foxl 2l和wnt 9 b分别在卵母细胞发育和性别决定和/或分化中的作用。我们的研究结果揭示了斑马鱼卵巢发育和功能的新见解，转录组谱将为未来的研究提供有价值的资源。
Zebrafish are an established research organism that has made many contributions to our understanding of vertebrate tissue and organ development, yet there are still significant gaps in our understanding of the genes that regulate gonad development, sex, and reproduction. Unlike the development of many organs, such as the brain and heart that form during the first few days of development, zebrafish gonads do not begin to form until the larval stage (≥5 days post-fertilization). Thus, forward genetic screens have identified very few genes required for gonad development. In addition, bulk RNA-sequencing studies that identify genes expressed in the gonads do not have the resolution necessary to define minor cell populations that may play significant roles in the development and function of these organs. To overcome these limitations, we have used single-cell RNA sequencing to determine the transcriptomes of cells isolated from juvenile zebrafish ovaries. This resulted in the profiles of 10,658 germ cells and 14,431 somatic cells. Our germ cell data represents all developmental stages from germline stem cells to early meiotic oocytes. Our somatic cell data represents all known somatic cell types, including follicle cells, theca cells, and ovarian stromal cells. Further analysis revealed an unexpected number of cell subpopulations within these broadly defined cell types. To further define their functional significance, we determined the location of these cell subpopulations within the ovary. Finally, we used gene knockout experiments to determine the roles of foxl2l and wnt9b for oocyte development and sex determination and/or differentiation, respectively. Our results reveal novel insights into zebrafish ovarian development and function, and the transcriptome profiles will provide a valuable resource for future studies.
影响因子：
14.9
作者：
Hsu RJ;Lin CY;Hoi HS;Zheng SK;Lin CC;Tsai HJ
通讯作者：
Tsai HJ