Live imaging of echinoderm embryos to illuminate evo-devo.
Live imaging of echinoderm embryos to illuminate evo-devo.
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DOI：
10.3389/fcell.2022.1007775
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发表时间：
2022
影响因子：
5.5
通讯作者：
Lyons, Deirdre C.
中科院分区：
文献类型：
--
作者：
Barone, Vanessa;Lyons, Deirdre C.
150多年来，棘皮动物胚胎一直是细胞和发育生物学的模型系统，这在很大程度上是因为它们的光学透明度。这些发现塑造了我们对受精、细胞分裂和细胞分化的理解，这是因为海胆卵和胚胎的透明性，可以直接观察细胞内结构。最近，海胆胚胎的实时成像，再加上荧光显微镜，已被证明是揭示上皮间充质转化，细胞迁移和原肠胚形成机制的关键。然而，实时成像主要是在海胆胚胎上进行的，而棘皮动物包括许多实验上易于处理的物种，这些物种在形态发生的关键方面存在有趣的变化，包括胚胎压实和囊胚形成机制的差异。对这种变异的研究不仅可以让我们了解棘皮动物的组织是如何形成的，还可以确定细胞形状、细胞-基质和细胞-细胞接触形成中的哪些变化更可能导致新的胚胎形状的进化。在这里，我们认为，适应活成像技术，以更多的棘皮动物物种将是根本的利用这种进化的方法来研究形态发生，因为它将允许测量动态细胞行为的差异-如细胞形状和细胞粘附的变化-物种之间。我们简要回顾现有的方法，活成像棘皮动物胚胎，并详细描述了我们如何适应这些方法，使长期活成像的几个物种，即海胆Lytechinus pictus和海星Patiria miniata和Patiriella regularis。我们概述了程序成功地标记，安装和图像早期胚胎10-16小时，从卵裂阶段到早期囊胚。我们表明，使用这些方法获得的数据允许随着时间的推移对单个细胞进行3D分割和跟踪，这是分析细胞形状和细胞接触在物种之间如何不同的第一步。这里提出的方法可以很容易地通过大多数细胞和发育生物学实验室，并适应成功地成像其他物种的早期胚胎，因此扩大了我们的理解形态发生的进化。
Echinoderm embryos have been model systems for cell and developmental biology for over 150 years, in good part because of their optical clarity. Discoveries that shaped our understanding of fertilization, cell division and cell differentiation were only possible because of the transparency of sea urchin eggs and embryos, which allowed direct observations of intracellular structures. More recently, live imaging of sea urchin embryos, coupled with fluorescence microscopy, has proven pivotal to uncovering mechanisms of epithelial to mesenchymal transition, cell migration and gastrulation. However, live imaging has mainly been performed on sea urchin embryos, while echinoderms include numerous experimentally tractable species that present interesting variation in key aspects of morphogenesis, including differences in embryo compaction and mechanisms of blastula formation. The study of such variation would allow us not only to understand how tissues are formed in echinoderms, but also to identify which changes in cell shape, cell-matrix and cell-cell contact formation are more likely to result in evolution of new embryonic shapes. Here we argue that adapting live imaging techniques to more echinoderm species will be fundamental to exploit such an evolutionary approach to the study of morphogenesis, as it will allow measuring differences in dynamic cellular behaviors - such as changes in cell shape and cell adhesion - between species. We briefly review existing methods for live imaging of echinoderm embryos and describe in detail how we adapted those methods to allow long-term live imaging of several species, namely the sea urchin Lytechinus pictus and the sea stars Patiria miniata and Patiriella regularis. We outline procedures to successfully label, mount and image early embryos for 10–16 h, from cleavage stages to early blastula. We show that data obtained with these methods allows 3D segmentation and tracking of individual cells over time, the first step to analyze how cell shape and cell contact differ among species. The methods presented here can be easily adopted by most cell and developmental biology laboratories and adapted to successfully image early embryos of additional species, therefore broadening our understanding of the evolution of morphogenesis.
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