A Cell Migration Tracking Tool Supports Coupling of Tissue Rotation to Elongation
A Cell Migration Tracking Tool Supports Coupling of Tissue Rotation to Elongation
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DOI：
10.1016/j.celrep.2017.09.083
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发表时间：
2017-10-17
期刊：
影响因子：
8.8
通讯作者：
Bilder, David
中科院分区：
文献类型：
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作者：
Chen, Dong-Yuan;Crest, Justin;Bilder, David
细胞迁移是形态发生和稳态不可或缺的。实时成像允许机械的见解，但长期观察可以改变正常的生物学，并且缺乏在体内跟踪运动而不受干扰的工具。我们在这里开发了一种称为M-TRAIL（实时和推断位置的矩阵标记技术）的工具，它揭示了固定组织中的迁移历史。使用过表达GFP标记的细胞外基质（ECM）成分的克隆，基于沉积在基底膜上的持久痕迹绘制运动轨迹。我们将M-TRAIL应用于果蝇卵泡旋转，比较体内和体外迁移动力学。在体内和体外测量的野生型（WT）卵泡旋转的速率、轨迹和停止是相同的，fat 2突变体的旋转失败也是如此。然而，携带细胞内截短的Fat 2的卵泡，以前报道缺乏旋转离体，实际上在体内旋转速度降低，从而重新验证了旋转是组织伸长所需的假设。M-TRAIL方法可用于跟踪和定量其他组织和生物体中的体内细胞运动。
Cell migration is indispensable to morphogenesis and homeostasis. Live imaging allows mechanistic insights, but long-term observation can alter normal biology, and tools to track movements in vivo without perturbation are lacking. We develop here a tool called M-TRAIL (matrix-labeling technique for real-time and inferred location), which reveals migration histories in fixed tissues. Using clones that overexpressGFP-tagged extracellularmatrix (ECM) components, motility trajectories are mapped based on durable traces deposited onto basement membrane. We applied M-TRAIL to Drosophila follicle rotation, comparing in vivo and ex vivo migratory dynamics. The rate, trajectory, and cessation of rotation in wild-type (WT) follicles measured in vivo and ex vivo were identical, as was rotation failure in fat2 mutants. However, follicles carrying intracellularly truncated Fat2, previously reported to lack rotation ex vivo, in fact rotate in vivo at a reduced speed, thus revalidating the hypothesis that rotation is required for tissue elongation. The M-TRAIL approach could be applied to track and quantitate in vivo cell motility in other tissues and organisms.