Ex vivo Follicle Rupture and in situ Zymography in Drosophila

Abstract

Ovulation, the process of releasing a mature oocyte from the ovary, is crucial for animal reproduction. In order for the process of ovulation to occur, a follicle must be fully matured and signaled to rupture from the ovary. During follicle rupture in both mammals and <i>Drosophila</i>, somatic follicle cells are enzymatically degraded to allow the oocyte to be liberated from the follicle. Here, we describe a detailed protocol of our newly developed <i>ex vivo</i> follicle rupture assay in <i>Drosophila</i>, which represents a first assay allowing direct quantification of follicles' capacity to respond to ovulation stimuli and rupture. This assay can be modified to stimulate rupture with other reagents (for example, ionomycin) or to query enzymatic activity (<i>in situ</i> zymography). In addition, this assay allows genetic or pharmacological screens to identify genes or small molecules regulating follicle rupture in <i>Drosophila</i>.