Democratizing embryo biomechanics: Development of robust and accessible methods to quantify sub-cellular mechanics in vivo

摘要： 现在人们普遍认为，对形态发生的任何全面理解都将 需要仔细量化体内的机械力。然而，我们面临着双重挑战 方法：首先，我们目前只在胚胎中拥有少量的亚细胞力学样本， 部分是因为这种分析受到对先进仪器的需要的限制。第二, 大多数力学性能分析不能提供足够的空间分辨率来量化 细胞离散子区域中的局部化机械不均一性。在这里，我们攻击这两个 发展稳健的方法对机械结构进行非侵入性量化的问题 细胞-细胞连接和细胞质的性质。提案将在以下方面开发这些工具 脊椎动物胚轴延长的背景，但这两种方法都应该被广泛使用， 只需要荧光记者和标准显微镜。该项目的成功将是1) 通过它们的高空间和时间分辨率以及它们的非 侵犯性和2)通过量化来提高我们的理解广度 胚胎的生物力学分析更容易为范围广泛的研究人员所接受。

Abstract: It is now widely recognized that any comprehensive understanding of morphogenesis will require careful quantification of mechanical forces in vivo. However, twin challenges stand in our way: First, we currently possess only a sparse sampling of subcellular mechanics in embryos, in part because such analyses are limited by the need for advanced instrumentation. Second, most analyses of mechanical properties do not provide sufficient spatial resolution to quantify localized mechanical heterogeneities in discrete sub-regions of the cell. Here, we attack both problems by developing robust methods for non-invasive quantification of the mechanical properties of both cell-cell junctions and the cytoplasm. The proposal will develop these tools in the context of vertebrate embryo axis elongation, but both methods should be widely accessible, requiring only fluorescent reporters and standard microscopes. Success of the project would 1) improve our depth of understanding through their high spatial and time resolution and their non- invasive nature and 2) improve our breadth of understanding by making quantitative biomechanical analysis in embryos far more accessible to a wide range of investigators.