Myocilin expression and apoptosis analysis in the immature (28 dpf) gonad of zebrafish KO for myoc. Analyses were performed on three consecutive tissue sections (1 to 3, upper left insert in panel A). Vasa immunolabeling was carried out on section one (green, A,B,E,F). Areas indicated by yellow rectangles in (A,E) are magnified in (B,F), respectively. Tissue section two was incubated with an anti-myocilin (TNT) antibody (green signals) to localise myocilin expression (C,G). Apoptosis was assessed in tissue section three using terminal dUTP nick-end labeling (TUNEL) of fragmented DNA (D,H). Arrows in (C) indicate myocilin immunoreactivity. Arrows and arrowheads in H indicate TUNEL-positive cells in gonadal tissue and in the outer intestinal wall, respectively. Blue and red signals correspond to DAPI nuclear staining and tissue autofluorescence, respectively. gn: gonad. in: intestine. ki: kidney. li: liver. sb: swimbladder. The vertical double arrow in (A) indicate de dorsovental axis (D: dorsal; V: ventral). The images are representative of the results observed in four fishes of each genotype. Negative controls of the juvenile gonad are shown in Figure S10 (tissue sections four to six).
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