title: Drosophila hemocyte migration: an in vivo assay for directional cell migration.
abstract: This protocol describes an in vivo assay for random and directed hemocyte migration in Drosophila. Drosophila is becoming an increasingly powerful model system for in vivo cell migration analysis, combining unique genetic tools with translucency of the embryo and pupa, which allows direct imaging and traceability of different cell types. In the assay we present here, we make use of the hemocyte response to epithelium wounding to experimentally induce a transition from random to directed migration. Time-lapse confocal microscopy of hemocyte migration in untreated conditions provides a random cell migration assay that allows identification of molecular mechanisms involved in this complex process. Upon laser-induced wounding of the thorax epithelium, a rapid chemotactic response changes hemocyte migratory behavior into a directed migration toward the wound site. This protocol provides a direct comparison of cells during both types of migration in vivo, and combined with recently developed resources such as transgenic RNAi, is ideal for forward genetic screens.
authors: Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Unidade de Morfogenese, Lisboa, Portugal.
authors: Carolina G A
authors: CG
authors: Moreira
authors: Jennifer C
authors: JC
authors: Regan
authors: Anna
authors: A
authors: Zaidman-Rémy
authors: Antonio
authors: A
authors: Jacinto
authors: Soren
authors: S
authors: Prag
mesh_terms: D000465:Algorithms; D000818:Animals; D030801:Animals, Genetically Modified; D054443:Cell Migration Assays / Q000379:methods*; D058948:Cell Tracking / Q000379:methods*; D002633:Chemotaxis; D004330:Drosophila / Q000166:cytology* / Q000235:genetics / Q000502:physiology; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics; D055785:Gene Knockdown Techniques; D006434:Hemocytes / Q000166:cytology* / Q000502:physiology; D007814:Larva / Q000166:cytology* / Q000235:genetics / Q000502:physiology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018613:Microscopy, Confocal; D034622:RNA Interference; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029721:Drosophila Proteins; D008164:Luminescent Proteins; D011994:Recombinant Proteins