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Fig. 4. Effect of olomoucine on F-actin and Src activation at the wound edge. Debridement wounds were made in corneas of wild-type mice, eyes were organ cultured for 12 hours, fixed and double-stained with rhodamine-phalloidin (F-actin) and antibody to activated Src (Src pY416). Stained tissues were dissected, whole mounted and examined by confocal microscopy. (A) F-Actin staining in the absence of olomoucine shows a sharp, regular wound edge, with actin staining at cell-cell boundaries and along the wound edge. (B) In the presence of olomoucine, the wound edge is disorganized and cells have separated from the epithelial cell sheet (arrows). F-Actin staining is seen in a broader band of cells along the wound edge than in untreated corneas. (C) Immunostaining of active Src (Src pY416) shows that Src is activated in a narrow band of cells adjacent to the wound edge. (D) In the presence of olomoucine, immunostaining of active Src suggests that inhibiting Cdk5 augments the activation of Src in many cells along the wound edge (arrows). Bar, 125 µm.
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