Culturing Drosophila Egg Chambers and Investigating Developmental Processes Through Live Imaging
- 1.9k Downloads
Drosophila oogenesis provides many examples of essential processes in development. A myriad of genetic tools combined with recent advances in culturing egg chambers ex vivo has revealed several surprising mechanisms that govern how this tissue develops, and which could not have been determined in fixed tissues. Here we describe a straightforward protocol for dissecting ovaries, culturing egg chambers, and observing egg development in real time by fluorescent microscopy. This technique is suitable for observation of early- or late-stage egg development, and can be adapted to study a variety of cellular, molecular, or developmental processes. Ongoing analysis of oogenesis in living egg chambers has tremendous potential for discovery of new developmental mechanisms.
KeywordsDrosophila melanogaster oogenesis Live imaging Ex vivo egg culturing Fluorescent microscopy Developmental genetics Cell migration Morphogenesis
This work was funded in part by a Department of Education Grant, Graduate Assistance in the Areas of National Need (GAANN) training fellowship (P200A120017), and by an NIGMS Initiative for Maximizing Student Development Grant (2 R25-GM55036), IMSD Meyerhoff Graduate Fellowship, to L.M. and a National Science Foundation CAREER Award (IOS-1054422) to M.S.G. We appreciate helpful comments on the manuscript from Dr. N. Sanchez-Alberola, G. Wunderlin, E. Desai, and D. DiMercurio, and we thank Dr. J. McDonald and Dr. A.C.C. Jang for sharing culturing information.
- 27.Weil TT, Parton RM, Davis I (2012) Preparing individual Drosophila egg chambers for live imaging. J Vis Exp (60): e3679Google Scholar
- 29.Pokrywka NJ (2013) Live imaging of GFP-labeled proteins in Drosophila oocytes. J Vis Exp (73): 50044Google Scholar
- 43.Cliffe A, Poukkula M, Rørth P (2009) Culturing Drosophila egg chambers and imaging border cell migration. Nat Protoc 10:289Google Scholar
- 46.Davis I, Parton RM (2006) Selection of appropriate imaging equipment and methodology for live cell imaging in Drosophila. CSH Protoc doi: 10.1101/pdb.ip21
- 47.Stonko D, Manning L, Starz-Gaiano M, Peercy B (2015) A force-based biophysical model of collective cell migration in a three-dimensional, heterogeneous environment. PLoS One. 10(4):e0122799Google Scholar