TY - JOUR
T1 - A protocol for culturing drosophila melanogaster stage 9 egg chambers for live imaging
AU - Prasad, Mohit
AU - Jang, Anna C.C.
AU - Starz-Gaiano, Michelle
AU - Melani, Mariana
AU - Montell, Denise J.
N1 - Funding Information:
ACKNOWLEDGMENTS This work was supported by National Institutes of Health grants GM46425 and GM73164 to D.J.M.
PY - 2007/10
Y1 - 2007/10
N2 - This protocol describes a method for the dissection of egg chambers from intact Drosophila females and culture conditions that permit live imaging of them, with a particular emphasis on stage 9. This stage of development is characterized by oocyte growth and patterning, outer follicle cell rearrangement and migration of border cells. Although in vitro culture of egg chambers of later developmental stages has long been possible, until recently stage 9 egg chambers could only be kept alive for short periods, did not develop normally, and border cell migration failed entirely. We have established culture conditions that support overall egg chamber development including border cell migration in vitro. This protocol makes possible direct observation of molecular and cellular dynamics in both wild-type and mutant egg chambers, and opens the door to testing of pharmacological inhibitors and the use of biosensors. The entire protocol takes ˜24 h while the preparation of egg chambers for live imaging requires only 15-20 min.
AB - This protocol describes a method for the dissection of egg chambers from intact Drosophila females and culture conditions that permit live imaging of them, with a particular emphasis on stage 9. This stage of development is characterized by oocyte growth and patterning, outer follicle cell rearrangement and migration of border cells. Although in vitro culture of egg chambers of later developmental stages has long been possible, until recently stage 9 egg chambers could only be kept alive for short periods, did not develop normally, and border cell migration failed entirely. We have established culture conditions that support overall egg chamber development including border cell migration in vitro. This protocol makes possible direct observation of molecular and cellular dynamics in both wild-type and mutant egg chambers, and opens the door to testing of pharmacological inhibitors and the use of biosensors. The entire protocol takes ˜24 h while the preparation of egg chambers for live imaging requires only 15-20 min.
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U2 - 10.1038/nprot.2007.363
DO - 10.1038/nprot.2007.363
M3 - Article
C2 - 17947988
AN - SCOPUS:38449092945
SN - 1754-2189
VL - 2
SP - 2467
EP - 2473
JO - Nature Protocols
JF - Nature Protocols
IS - 10
ER -