Abstract
Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing late stage egg chambers was long feasible, stage 9 egg chambers survived only briefly outside the female body. We identified culture conditions that support stage 9 egg chamber development and sustain complete migration of border cells ex vivo. This protocol enables one to compare the dynamics of egg chamber development in wild type and mutant egg chambers using time-lapse microscopy and taking advantage of a multiposition microscope with a motorized imaging stage. In addition, this protocol has been successfully used in combination with fluorescence resonance energy transfer biosensors, photo-activatable proteins, and pharmacological agents and can be used with widefield or confocal microscopes in either an upright or inverted configuration.
This volume includes electronic supplemental material that will appear on the website http://extras.springer.com/wells.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Ilina, O., and Friedl, P. (2009) Mechanisms of collective cell migration at a glance. J. Cell Sci. 122, 3203–8.
Aman, A., and Piotrowski, T. (2010) Cell migration during morphogenesis. Dev. Biol. 341, 20–33.
Weijer, C.J. (2009) Collective cell migration in development. J. Cell Sci. 122, 3215–23.
López-Schier, H. (2010) Fly fishing for collective cell migration. Curr. Opin. Genet. Dev. 20, 428–32.
Rørth, P. (2009) Collective cell migration. Annu. Rev. Cell Dev. Biol. 25, 407–29.
Spradling, A.C. (1993) Developmental Genetics of Oogenesis. In The Development of Drosophila melanogaster. M. Bate and A. Martinez-Arias, eds. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press), pp. 1–70.
King, R.C. (1970) Ovarian Development in Drosophila melanogaster (Academic Press, New York).
Edgar, B.A., and Orr-Weaver, T.L. (2001) Cell. 105, 297–306.
Ruohola, H., Bremer, K.A., Baker, D., Swedlow, J.R., Jan, L.Y., and Jan, Y.N. (1991) Role of neurogenic genes in establishment of follicle cell fate and oocyte polarity during oogenesis in Drosophila. Cell. 66, 433–449.
Silver, D.L., and Montell, D.J. (2001) Paracrine signaling through the JAK/STAT pathway activates invasive behavior of ovarian epithelial cells in Drosophila. Cell. 107, 831–41.
Montell, D.J., Rørth, P., and Spradling, A.C. (1992) Slow border cells, a locus required for a developmentally regulated cell migration during oogenesis, encodes Drosophila C/EBP. Cell. 71, 51–62.
Starz-Gaiano, M., Melani, M., Wang, X., Meinhardt, H., and Montell, D.J. (2008) Feedback inhibition of Jak/STAT signaling by apontic is required to limit an invasive cell population. Dev. Cell. 14, 726–38.
Prasad, M., and Montell, D. J. (2007) Cellular and Molecular Mechanisms of Border Cell Migration Analyzed Using Time-lapse Live-cell Imaging. Dev. Cell. 12, 997–1005.
Bianco, A., Poukkula, M., Cliffe, A., Mathieu, J., Luque, C. M., Fulga, T. A., and Rorth, P. (2007) Two distinct modes of guidance signalling during collective migration of border cells. Nature. 448, 362–65.
Duchek, P., Somogyi, K., Jekely, G., Beccari, S., and Rorth, P. (2001) Guidance of cell migration by the drosophila PDGF/VEGF receptor. Cell. 107, 17–26.
McDonald, J. A., Pinheiro, E. M., Kadlec, L., Schupbach, T., and Montell, D. J. (2006) multiple EGFR ligands participate in guiding migrating border cells. Dev. Biol. 296, 94–103.
Duchek, P., and Rorth, P. (2001) Guidance of cell migration by EGF receptor signaling during Drosophila oogenesis. Science. 291, 131–33.
Prasad, M., Jang, A.C., Starz-Gaiano, M., Melani, M., and Montell, D.J. (2007) A protocol for culturing Drosophila melanogaster stage 9 egg chambers for live imaging. Nat. Protoc. 2, 2467–73.
Wang, X., He, L., Wu, Y.I., Hahn, K.M., and Montell, D.J. (2010) Light-mediated activation reveals a key role for Rac in collective guidance of cell movement in vivo. Nat. Cell Biol. 12, 591–7.
Horne-Badovinac, S., and Bilder, D. (2005) Mass transit: epithelial morphogenesis in the Drosophila egg chamber. Dev. Dyn. 232, 559–72.
Grunert, S. and St Johnston, D. (1996) RNA localization and the development of asymmetry during Drosophila oogenesis. Curr. Opin. Genet. Dev. 6, 395–402.
Hudson, A.M., and Cooley, L. (2002) Understanding the function of actin-binding proteins through genetic analysis of Drosophila oogenesis. Annu. Rev. Genet. 36, 455–88.
Rørth, P., Szabo, K., Bailey, A., Laverty, T., Rehm, J., Rubin, G.M., Weigmann, K., Milán, M, Benes, V., Ansorge, W., and Cohen, S.M. (1998) Systematic gain-of-function genetics in Drosophila. Development. 125, 1049–57.
Wang, X., Bo, J., Bridges, T., Dugan, K. D., Pan, T. C., Chodosh, L. A., and Montell, D. J. (2006) Analysis of cell migration using whole-genome expression profiling of migratory cells in the Drosophila ovary. Dev. Cell. 10, 483–95.
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
Prasad, M., Wang, X., He, L., Montell, D.J. (2011). Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement. 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_19
Download citation
DOI: https://doi.org/10.1007/978-1-61779-207-6_19
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-206-9
Online ISBN: 978-1-61779-207-6
eBook Packages: Springer Protocols