title: Drosophila pupal macrophages--a versatile tool for combined ex vivo and in vivo imaging of actin dynamics at high resolution.
abstract: Molecular understanding of actin dynamics requires a genetically traceable model system that allows live cell imaging together with high-resolution microscopy techniques. Here, we used Drosophila pupal macrophages that combine many advantages of cultured cells with a genetic in vivo model system. Using structured illumination microscopy together with advanced spinning disk confocal microscopy we show that these cells provide a powerful system for single gene analysis. It allows forward genetic screens to characterize the regulatory network controlling cell shape and directed cell migration in a physiological context. We knocked down components regulating lamellipodia formation, including WAVE, single subunits of Arp2/3 complex and CPA, one of the two capping protein subunits and demonstrate the advantages of this model system by imaging mutant macrophages ex vivo as well as in vivo upon laser-induced wounding.
authors.affiliation: Institute for Neurobiology, University of Münster, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Sander
authors.forename: Anna Julia
authors.initials: AJ
authors.lastname: Squarr
authors.forename: Benjamin
authors.initials: B
authors.lastname: Risse
authors.forename: Xiaoyi
authors.initials: X
authors.lastname: Jiang
authors.forename: Sven
authors.initials: S
authors.lastname: Bogdan
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D004330:Drosophila / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D023421:Models, Animal; D011679:Pupa / Q000166:cytology / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins
keywords: Cell motility; Cell shape; Drosophila; F-actin; Live cell imaging; Macrophages; RNAi; Structured illumination microscopy