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

Cell Motil Cytoskeleton. 2006 Jul;63(7):415-22. doi: 10.1002/cm.20133.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / pharmacology
  • Animals
  • Chemotaxis / drug effects
  • Chemotaxis / physiology*
  • Embryo, Nonmammalian
  • Enzyme Inhibitors / pharmacology
  • Green Fluorescent Proteins / metabolism
  • Inflammation / physiopathology
  • Intracellular Signaling Peptides and Proteins
  • Macrophages / drug effects
  • Macrophages / physiology*
  • Microscopy, Video
  • Microtubules / drug effects
  • Microtubules / metabolism*
  • Nocodazole / pharmacology
  • Protein-Serine-Threonine Kinases / antagonists & inhibitors
  • Pyridines / pharmacology
  • Wounds and Injuries / etiology
  • Wounds and Injuries / physiopathology
  • Yolk Sac / cytology*
  • Yolk Sac / injuries*
  • Zebrafish / embryology*
  • Zebrafish / physiology
  • rho-Associated Kinases

Substances

  • Amides
  • Enzyme Inhibitors
  • Intracellular Signaling Peptides and Proteins
  • Pyridines
  • Y 27632
  • Green Fluorescent Proteins
  • Protein-Serine-Threonine Kinases
  • rho-Associated Kinases
  • Nocodazole