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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Luster, A. D., Alon, R., and von Andrian, U. H. (2005) Immune cell migration in inflammation: present and future therapeutic targets, Nat Immunol 6, 1182–1190.
Yamaguchi, H., Wyckoff, J., and Condeelis, J. (2005) Cell migration in tumors, Current Opinion in Cell Biology 17, 559–564.
Rørth, P. (2009) Collective Cell Migration, Annual Review of Cell and Developmental Biology 25, 407–429.
Petrie, R. J., Doyle, A. D., and Yamada, K. M. (2009) Random versus directionally persistent cell migration, Nat Rev Mol Cell Biol 10, 538–549.
Chen, H. C. (2005) Boyden chamber assay, Methods Mol Biol 294, 15–22.
Zicha, D., Dunn, G., and Jones, G. (1997) Analyzing chemotaxis using the Dunn direct-viewing chamber, Methods Mol Biol 75, 449–457.
Fraley, S. I., Feng, Y., Krishnamurthy, R., Kim, D.-H., Celedon, A., Longmore, G. D., and Wirtz, D. (2010) A distinctive role for focal adhesion proteins in three-dimensional cell motility, Nat Cell Biol 12, 598–604.
Gaggioli, C., Hooper, S., Hidalgo-Carcedo, C., Grosse, R., Marshall, J. F., Harrington, K., and Sahai, E. (2007) Fibroblast-led collective invasion of carcinoma cells with differing roles for RhoGTPases in leading and following cells, Nat Cell Biol 9, 1392–1400.
Brand, A. H., and Perrimon, N. (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes, Development 118, 401–415.
Duffy, J. B. (2002) GAL4 system in drosophila: A fly geneticist’s swiss army knife, Genesis 34, 1–15.
Stramer, B., and Wood, W. (2009) Inflammation and Wound Healing in Drosophila, Methods Mol Biol, pp 137–149.
Xu, T., and Rubin, G. M. (1993) Analysis of genetic mosaics in developing and adult Drosophila tissues, Development 117, 1223–1237.
Wood, W., and Jacinto, A. (2007) Drosophila melanogaster embryonic haemocytes: masters of multitasking, Nat Rev Mol Cell Biol 8, 542–551.
Bruckner, K., Kockel, L., Duchek, P., Luque, C. M., Rorth, P., and Perrimon, N. (2004) The PDGF/VEGF Receptor Controls Blood Cell Survival in Drosophila, Developmental Cell 7, 73–84.
Bainbridge, S., and Bownes, M. (1981) Staging the metamorphosis of Drosophila melanogaster, J Embryol Exp Morph 66, 57–80
Acknowledgments
The authors would like to acknowledge Marco Antunes for suggestions on hemocytes, Jan Stühmer for suggestions on cell tracking, and Luis Almeida for input on analysis. This work was supported by FCT PTDC/SAU-OBD/101259/2008 (S.P.) and SFRH/BD/62345/2009 (C.G.A.M.), ARC and FCT SFRH/BPD/44613/2008 fellowships (A.Z.R.), and EMBO ALTF 178-2009 (J.R.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Moreira, C.G.A., Regan, J.C., Zaidman-Rémy, A., Jacinto, A., Prag, S. (2011). Drosophila Hemocyte Migration: An In Vivo Assay for Directional Cell Migration. In: Wells, C., Parsons, M. (eds) Cell Migration. Methods in Molecular Biology, vol 769. Humana Press. https://doi.org/10.1007/978-1-61779-207-6_17
Download citation
DOI: https://doi.org/10.1007/978-1-61779-207-6_17
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-206-9
Online ISBN: 978-1-61779-207-6
eBook Packages: Springer Protocols