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A Cell Migration Tracking Tool Supports Coupling of Tissue Rotation to Elongation

机译:细胞迁移跟踪工具支持组织旋转与伸长的耦合

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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 overexpress GFP-tagged extracellular matrix (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.
机译:细胞迁移对于形态发生和体内稳态是必不可少的。实时成像可以提供机械方面的见解,但是长期观察可以改变正常生物学,因此缺乏在不干扰的情况下跟踪体内运动的工具。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,该工具可揭示固定组织中的迁移历史。使用过度表达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,比较了体内和离体的迁徙动力学。在体内和离体测量的野生型(WT)卵泡的速率,轨迹和旋转停止是相同的,fat2突变体中的旋转失败也是相同的。然而,先前报道过的缺乏胞内旋转的胞内截短的Fat2的卵泡实际上在体内以降低的速度旋转,因此重新验证了组织伸长需要旋转的假设。 M-TRAIL方法可用于跟踪和定量其他组织和生物体内的体内细胞运动。

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