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The dynamics of epidermal stratification during post-larval development in zebrafish.

机译:斑马鱼幼虫发育过程中表皮分层的动态。

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Background: The epidermis, as a defensive barrier, is a consistent trait throughout animal evolution. During post-larval development, the zebrafish epidermis thickens by stratification or addition of new cell layers. Epidermal basal stem cells, expressing the transcription factor p63, are known to be involved in this process. Zebrafish post-larval epidermal stratification is a tractable system to study how stem cells participate in organ growth. Methods: We used immunohistochemistry, in combination with EdU cell proliferation detection, to study zebrafish epidermal stratification. For this procedure, we selected a window of post-larval stages (5-8 mm of standard length or SL, which normalizes age by size). Simultaneously, we used markers for asymmetric cell division and the Notch signaling pathway. Results: We found that epidermal stratification is the consequence of several events, including changes in cell shape, active cell proliferation and asymmetrical cell divisions. We identified a subset of highly proliferative epidermal cells with reduced levels of p63, which differed from the basal stem cells with high levels of p63. Additionally, we described different mechanisms that participate in the stratification process, including the phosphorylation of p63, asymmetric cell division regulated by the Par3 and LGN proteins, and expression of Notch genes.
机译:背景:表皮作为一种防御屏障,在动物进化过程中是一种一致的特征。在仔鱼后期发育期间,斑马鱼表皮通过分层或添加新的细胞层而增厚。表皮基底干细胞表达转录因子p63,已知参与这一过程。斑马鱼幼虫后表皮分层是研究干细胞如何参与器官生长的一个易于处理的系统。方法:采用免疫组织化学方法,结合EdU细胞增殖检测,研究斑马鱼表皮分层。对于这个过程,我们选择了一个幼虫后期窗口(标准长度为5-8mm或SL,通过大小使年龄正常化)。同时,我们使用不对称细胞分裂和Notch信号通路的标记。结果:我们发现表皮分层是多种事件的结果,包括细胞形状的变化、活跃的细胞增殖和不对称的细胞分裂。我们鉴定了一组p63水平降低的高度增殖表皮细胞,这与p63水平高的基底干细胞不同。此外,我们还描述了参与分层过程的不同机制,包括p63的磷酸化、由Par3和LGN蛋白调节的不对称细胞分裂,以及Notch基因的表达。

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