Effect of Fertilization on the distribution of active Src-family PTKs near the point of sperm entry.
Eggs were fixed before and at 30 seconds or 60seconds post-insemination. The chorion was permeabilized by proteolysis but not removed so that the micropyle could be accurately visualized. Eggs were immunolabeled with the clone 28 antibody to detect active Src-family PTKs followed by goat anti-mouse Alexa 488 (green). Sperm chromatin was labeled with DAPI (blue) and in some cases, actin was detected by binding of alexa Fluor 568-phalloidin (red). Conditions of illumination and fluorescent signal amplification were adjusted to optimize visualization of the pattern of immunofluorescence and are not intended for comparison of the intensity of immunofluorescence. All eggs were oriented with the micropyle (black arrows) toward the top of each panel. The sperm head, stained with DAPI, is indicated by a white arrow. Magnification is indicated by the bar which represents 100μm.