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Imaging macrophage chemotaxis in vivo: studies of microtubule function in zebrafish wound inflammation.

机译:体内成像的巨噬细胞趋化性:斑马鱼伤口炎症中微管功能的研究。

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摘要

The inflammatory response is one of the most dramatic examples of directed cell movement in nature. Inflammation is triggered at the site of injury and results in the migration of immune cells to the site to protect the host from infection. We have devised an in vivo inflammation assay using translucent zebrafish embryos, which allow live imaging and pharmacological manipulation of macrophage chemotaxis to wounds inflicted with a laser. Using this assay, we test the role of the microtubule cytoskeleton in macrophage chemotaxis in vivo using nocodazole to disrupt microtubule polymerization. We find that de-stabilisation of microtubules with nocodazole impairs macrophage recruitment to wounds, but that addition of the Rho kinase inhibitor Y-27632 suppresses these effects and restores the recruitment of macrophages to wounds. Taken together, these results suggest that destabilizing microtubules activates Rho kinase and that this increase in Rho kinase activity interferes with leukocyte recruitment in vivo.
机译:炎症反应是自然界中定向细胞运动的最引人注目的例子之一。炎症在受伤部位被触发,并导致免疫细胞迁移到该部位以保护宿主免受感染。我们设计了一种使用半透明斑马鱼胚胎的体内炎症测定方法,该方法可对巨噬细胞的趋化性进行实时成像和药理处理,以应对激光造成的伤口。使用这种测定法,我们使用诺考达唑破坏微管聚合反应,测试了微管细胞骨架在体内巨噬细胞趋化作用中的作用。我们发现用诺考达唑使微管失去稳定作用会损害巨噬细胞向伤口的募集,但是添加Rho激酶抑制剂Y-27632会抑制这些作用并恢复巨噬细胞向伤口的募集。综上所述,这些结果表明不稳定的微管激活了Rho激酶,而Rho激酶活性的这种增加会干扰体内白细胞的募集。

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