基于单细胞转录组测序分析构建花斑裸鲤三种造血器官细胞图谱
作者:
作者单位:

1.青海大学省部共建三江源生态与高原农牧业国家重点实验室 西宁 810016;2.青海大学生态环境工程学院 西宁 810016;3 .青海大学农牧学院动物科学系 西宁 810016

作者简介:

寇若彬,女,硕士研究生;研究方向:生态学;E-mail:krb18009712507@163.com。

基金项目:

青海省自然科学基金项目(No. 2020-ZJ-907),国家自然科学基金项目(No. 31960127);


Generating Cellular Atlases of Three Haematopoietic Organs of the Gymnocypris eckloni using Single-Cell RNA Sequencing
Author:
Affiliation:

1.State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining 810016; 2.College of Eco-Environmental Engineering, Qinghai University, Xining 810016; 3.Animal Science Department of Agriculture and Animal Husbandry College, Qinghai University, Xining 810016, China

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    摘要:

    利用单细胞转录组测序技术在细胞水平探究花斑裸鲤(Gymnocypris eckloni)肾、肝和脾三种造血器官中的细胞组成类型,系统描绘花斑裸鲤这三种造血器官的细胞图谱,为进一步研究花斑裸鲤造血系统发育,尤其是红细胞发育积累科学数据。研究方法包括花斑裸鲤肾、肝和脾三种造血器官的单细胞悬液制备、单细胞RNA-seq文库构建、测序以及降维聚类分析、细胞注释、差异基因分析和功能富集分析。结果表明,花斑裸鲤三种造血器官内存在类型丰富的细胞,包括红细胞、中性粒细胞、T细胞、B细胞、树突状细胞、上皮细胞、巨噬细胞、血管内皮细胞、肌细胞、成纤维细胞、单核细胞和神经细胞。在三种器官中细胞分布存在较大的异质性:脾中参与发挥红细胞功能的细胞群分布占比较大;肾中参与免疫反应的细胞群分布占比较大;肝中血管内皮细胞和肌细胞分布占比较大。在功能方面,花斑裸鲤与多数硬骨鱼类相似,它的肾、脾和肝不仅参与造血及红细胞的发育,而且对于免疫系统的发育和应答等过程也发挥一定作用。本研究从单细胞角度对花斑裸鲤造血系统发育研究进行了一定的补充与完善,展现了三种造血器官中细胞分布的特征与差异,对于后续进一步预测花斑裸鲤的红细胞发育路径提供了一定的研究基础。

    Abstract:

    [Objectives] Single-cell RNA sequencing was used to investigate the cellular composition of the three hematopoietic organs-kidney, liver, and spleen of Gymnocypris eckloni. The study aims to systematically map the cellular composition of these organs, and to accumulate scientific data for the study of the haematopoietic system, particularly erythropoiesis, in this species. [Methods] Single-cell suspensions were prepared and sequenced using single cell RNA sequencing software to clarify the characteristics and differences in the distribution of cells in the haematopoietic organs and to construct a preliminary cellular atlas of the haematopoietic organs of G. eckloni. [Results] Cells within the three organs of G. eckloni were initially classified into 16 subpopulations based on the sequencing results (Fig. 2). This, combined with the expression of the top 20 differentially expressed genes and known marker genes for each cell population, identified a variety of cell types. These included erythrocytes, neutrophils, T cells, B cells, dendritic cells, epithelial cells, macrophages, vascular endothelial cells, myoblasts, fibroblasts, monocytes and neuronal cells. There is considerable heterogeneity in the distribution of cells in the three organs (Fig. 4). The G. eckloni had a relatively large distribution of cell populations involved in erythrocyte functions, the kidney had a relatively large distribution of cell populations involved in the immune response, and the liver had a relatively large distribution of vascular endothelial cells and myocytes. [Conclusion] The similarity to the haematopoietic cell types of the Zebrafish Danio rerio demonstrates the high conservation of haematopoietic cell development and evolution. According to functional enrichment analysis, G. eckloni, like most scleractinian fishes, has kidneys, spleen and liver, which are not only involved in haematopoiesis and erythropoiesis, but also play roles in immune system development, response and other processes. Meanwhile, the present study complements and improves the study of the haematopoietic phylogeny of G. eckloni from a single-cell perspective, shows the characteristics and differences of cell distribution in different haematopoietic tissues, and provides a certain research basis for further predicting the developmental path of erythrocytes in G. eckloni.

    参考文献
    Agius C. 1980. Phylogenetic development of melano-macrophage centres in fish. Journal of Zoology, 191(1):11–31.
    Akashi K, Traver D, Miyamoto T, et al. 2000. A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature, 404(6774):193–197.
    Alberti-Servera L, von Muenchow L, Tsapogas P, et al. 2017. Single-cell RNA sequencing reveals developmental heterogeneity among early lymphoid progenitors. The EMBO Journal, 36(24):3619–3633.
    Dalmo R A, Ingebrigtsen K, B?gwald J. 1997. Non-specific defence mechanisms in fish, with particular reference to the reticuloendothelial system (RES). Journal of Fish Diseases, 20(4):241–273.
    Dong F, Hao S, Zhang S, et al. 2020. Differentiation of transplanted haematopoietic stem cells tracked by single-cell transcriptomic analysis. Nature Cell Biology, 22(6):630–639.
    Ferguson H W, Claxton M J, Moccia R D, et al. 1982. The quantitative clearance of bacteria from the bloodstream of rainbow trout (Salmogairdneri). Veterinary Pathology, 19(6):687–699.
    Giladi A, Paul F, Herzog Y, et al. 2018. Single-cell characterization of haematopoietic progenitors and their trajectories in homeostasis and perturbed haematopoiesis. Nature Cell Biology, 20(7):836–846.
    Gordeuk V R, Stockton D W, Prchal J T. 2005. Congenital polycythemias/erythrocytoses. Haematologica, 90(1):109–116.
    Hinton D, Lauren J. 1990. Integrative histopathological approaches to detecting effects of environmental stressors on fishes. Transactions of the American Fisheries Society, 8:51–66.
    Kaattari S L, Irwin M J. 1985. Salmonid spleen and anterior kidney harbor populations of lymphocytes with different B cell repertoires. Developmental & Comparative Immunology, 9(3):433–444.
    Lorenzo F, Dalla Venezia N, Morlé L, et al. 1994. Protein 4.1 deficiency associated with an altered binding to the spectrin-actin complex of the red cell membrane skeleton. Journal of Clinical Investigation, 94(4):1651–1656.
    Nestorowa S, Hamey F K, Pijuan Sala B, et al. 2016. A single-cell resolution map of mouse hematopoietic stem and progenitor cell differentiation. Blood, 128(8):e20–e31.
    Paul F, Arkin Y, Giladi A, et al. 2015. Transcriptional heterogeneity and lineage commitment in myeloid progenitors. Cell, 163(7):1663–1677.
    Tusi B K, Wolock S L, Weinreb C, et al. 2018. Population snapshots predict early haematopoietic and erythroid hierarchies. Nature, 555(7694):54–60.
    Villani A C, Satija R, Reynolds G, et al. 2017. Single-cell RNA-seq reveals new types of human blood dendritic cells, monocytes, and progenitors. Science, 356(6335):eaah4573.
    Watcham S, Kucinski I, Gottgens B. 2019. New insights into hematopoietic differentiation landscapes from single-cell RNA sequencing. Blood, 133(13):1415–1426.
    Wilson N K, Kent D G, Buettner F, et al. 2015. Combined single-cell functional and gene expression analysis resolves heterogeneity within stem cell populations. Cell Stem Cell, 16(6):712–724.
    Xia J, Kang Z X, Xue Y Y, et al. 2021. A single-cell resolution developmental atlas of hematopoietic stem and progenitor cell expansion in zebrafish. Proceedings of the National Academy of Sciences of the United States of America, 118(14):e2015748118.
    Xie X W, Liu M Y, Zhang Y W, et al. 2020. Single-cell transcriptomic landscape of human blood cells. National Science Review, 8(3):nwaa180.
    陈涛. 2022. 单细胞整合分析解析小鼠早期造血发育. 昆明:昆明理工大学硕士学位论文.
    陈仲扬, 马艳妮, 余佳. 2022. 基于单细胞转录组测序分析描绘人类早期胚胎红细胞发育图谱. 基础医学与临床, 42(5):776–781.
    初小雅. 2016. 斑马鱼胸腺和头肾的结构特征及其组织样品的制备程序. 南京:南京农业大学硕士学位论文.
    韩光明, 毕建花, 张家宏, 等. 2015. 间氨基苯甲酸乙酯甲磺酸盐对泥鳅的麻醉效果. 广东海洋大学学报, 35(4):17–23.
    黄英毅. 2022. 基于单细胞测序的斑马鱼肝脏细胞异质性研究. 上海:上海海洋大学硕士学位论文.
    李华, 邓林. 2014. 黄河花斑裸鲤肌肉中矿物质元素测定与营养评价. 四川动物, 33(6):923–925.
    李玉邦. 2023. 单细胞RNA测序分析揭示了斑马鱼头肾中的造血通路. 济南:山东师范大学硕士学位论文.
    马燕梅, 林树根, 王全溪, 等. 2008. 花鲈头肾的显微结构和超显微结构. 福建农林大学学报:自然科学版, 37(2):190–193.
    祁得林. 2009. 黄河上游花斑裸鲤Cyt b基因的序列变异和遗传多样性. 动物学研究, 30(3):255–261.
    王军平, 陈乃成. 2021. 单细胞测序在造血系统研究中的应用与展望. 第三军医大学学报, 43(17):1603–1610.
    王胜涛, 朱浩然, 徐淑娟, 等. 2023. 基于单细胞测序数据的骨肉瘤预后相关细胞亚群特征分析. 中国医学科学院学报, 45(5):773–782.
    王新栋. 2018. 斑马鱼脾脏、心脏与体肾的微细结构及其样品制备标准化. 南京:南京农业大学硕士学位论文.
    武云飞, 谭齐佳. 1991. 青藏高原鱼类区系特征及其形成的地史原因分析. 动物学报, 37(2):135–152.
    武云飞, 吴翠珍. 1982. 青藏高原鱼类. 成都:四川科学技术出版社.
    岳兴建, 张耀光, 敖磊, 等. 2008. 南方鲇前肾的发生及头肾的形成. 西南大学学报:自然科学版, (7):110–116.
    张成鹏. 2022. 单细胞测序技术及其应用综述. 河南科学, 40(9):1390–1397.
    张向辉. 2022. 单细胞测序探究斑马鱼红细胞发育及细胞核保留机制. 大连:大连海洋大学硕士学位论文.
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寇若彬,刘丹,高强,晁燕,张存芳,聂苗苗,贾军梅,谭瑾,郭守全,姚占雯,祁得林.2025.基于单细胞转录组测序分析构建花斑裸鲤三种造血器官细胞图谱.动物学杂志,60(1):69-79.

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  • 收稿日期:2023-12-19
  • 在线发布日期: 2025-03-04