The shape, position and nuclei number of lens fiber nuclei are affected in the vov mutant. (A) Color key for lens epithelial cells and lens fiber cells. From 36 to 72 hpf, the margin of Pax6-positive lens epithelium (red) is associated with the retinal CMZ (light blue). Lens fiber cells are defined as Pax6-negative lens cells (green). The interface between the anterior and posterior half of the lens sphere is indicated by a dotted line. Index of nucleus flatness (Inf) is calculated as the ratio of the maximum (Lmax) to the minimum (Lmin) nuclear thickness. Inf=1.0-2.0, dark blue; 2.0-3.0, light blue; 3.0-4.0, green; 4.0-5.0, yellow; more than 5.0, red. (B-I) EdU incorporation (green) and DAPI labeling (red) of wild-type (B-E) and the vov mutant (F-I) zebrafish lens. Asterisk shows non-specific EdU signals (I). (J,K) Nuclear staining (upper panels) of wild-type (J) and the vov mutant (K) lens with DAPI. Inf is plotted to individual nuclei of lens fiber cells on the lower row of images. The degree of flatness is indicated by the height of bars and their color (see A). Dotted lines indicate the interface between the anterior and posterior halves. (L) The percentage of EdU-positive cells relative to the total number of lens epithelial cells in wild type (light blue bars) and the vov mutant (red bars). (M) The number of lens fiber cell nuclei in the wild type (light blue bars) and vov mutant (red bars). (N) Index of nuclear anterior localization (Ina) of the wild type and vov mutant. (O) The Inf of individual lens fiber cell nuclei in the wild type and vov mutant. Scale bars: 50 µm.
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