Title | Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement. |
Publication Type | Journal Article |
Year of Publication | 2015 |
Authors | Prasad M, Wang X, He L, Cai D, Montell DJ |
Journal | Methods Mol Biol |
Volume | 1328 |
Pagination | 89-97 |
Date Published | 2015 |
ISSN | 1940-6029 |
Keywords | Animals, Cell Movement, Drosophila melanogaster, Female, Microscopy, Fluorescence, Molecular Biology, Oogenesis, Organ Culture Techniques, Ovum |
Abstract | <p>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.</p> |
DOI | 10.1007/978-1-4939-2851-4_6 |
Alternate Journal | Methods Mol. Biol. |
PubMed ID | 26324431 |
PubMed Central ID | PMC4762686 |
Grant List | R01 GM073164 / GM / NIGMS NIH HHS / United States GM73164 / GM / NIGMS NIH HHS / United States |