DFG-Center for Regenerative Therapies Dresden, Cluster of Excellence, Technische Universität Dresden, Dresden, Germany.
Paul Langerhans Institute Dresden of the Helmholtz Zentrum München at the University Hospital and Faculty of Medicine Carl Gustav Carus of Technische Universität Dresden, Dresden, Germany.
Methods Mol Biol. 2020;2128:159-179. doi: 10.1007/978-1-0716-0385-7_12.
During embryogenesis, beta-cells arise from the dorsal and ventral bud originating in the endoderm germ layer. As the animal develops to adulthood, the beta-cell mass dramatically increases. The expansion of the beta-cell population is driven by cell division among the embryonic beta-cells and supplanted by neogenesis from post-embryonic progenitors. Here, we describe a protocol for multicolor clonal analysis in zebrafish to define the contribution of individual embryonic beta-cells to the increase in cell numbers. This technique provides insights into the proliferative history of individual beta-cells in an islet. This insight helps in defining the replicative heterogeneity among individual beta-cells during development. Additionally, the ability to discriminate individual cells based on unique color signatures helps quantify the volume occupied by beta-cells and define the contribution of cellular size to the beta-cell mass.
在胚胎发生过程中，β细胞起源于内胚层芽的背侧和腹侧。随着动物发育到成年，β细胞质量显著增加。β细胞群体的扩张是由胚胎β细胞之间的细胞分裂驱动的，并由胚胎后祖细胞的新生所取代。在这里，我们描述了一种在斑马鱼中进行多色克隆分析的方案，以确定单个胚胎β细胞对细胞数量增加的贡献。该技术提供了对胰岛中单个β细胞增殖历史的深入了解。这种见解有助于在发育过程中定义个体β细胞之间的复制异质性。此外，基于独特颜色特征区分单个细胞的能力有助于量化β细胞占据的体积，并定义细胞大小对β细胞质量的贡献。
