Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement.
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 wide-field or confocal microscopes in either an upright or an inverted configuration.
Department of Biological Chemistry, Johns Hopkins School of Medicine, Baltimore, MD, 21205, USA.
Mohit
M
Prasad
Xiaobo
X
Wang
Li
L
He
Danfeng
D
Cai
Denise J
DJ
Montell
D000818:Animals; D002465:Cell Movement / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics; D005260:Female; D008856:Microscopy, Fluorescence; D008967:Molecular Biology / Q000379:methods*; D009866:Oogenesis / Q000235:genetics; D009924:Organ Culture Techniques / Q000379:methods*; D010063:Ovum / Q000254:growth & development*