File:Identification-and-Functional-Analysis-of-Healing-Regulators-in-Drosophila-pgen.1004965.s024.ogv

English: Live imaging of imaginal disc healing in vitro . Imaginal discs were cultured in modified medium during 16–18 hours. Membranes are shown in red (FM4–64) and puc expression is shown in green (pucE69-Gal4 A; UAS-GFP). Few hours (6) after wounding a clear shortening of the wound edge while cells at the wound margin showed actin-rich protrusions more abundant in the close proximity of the vertex of the healing wound is observed. In addition to these filopodia, we detected a rapid actin cytoskeleton reorganization that occurred in both, the columnar and the peripodial epithelium with the formation of a purse-string. 12 hours after cutting the discs, the size of the gap is considerably reduced and by 16–18 hours the incised discs have zipped and sealed the gap. Hence, healing does undergo a slightly slower kinetics than that showed for implanted discs (12 hours for completion). Also, in contrast to in vivo cultured discs, a mass of cell debris was frequently observed, attached to the wound vertex that was actively extruded upon edges apposition. puc expression initiates after 2 hours in culture in scattered cells at the wound edges. At 7 hours post wounding most of the cells at the wound edges express puc. This expression develops further and reaches a maximum at 15 hours of cultured when several rows of cells decorated the almost fully healed region.