Figure 7. Whole lens staining for F-actin (phalloidin, green) and nuclei (DAPI, red) in 4-month-old and 18-month-old lenses. (A) The maximum intensity projection of the anterior lens epithelium and underlying fibers in the 4-month-old lens shows evenly distributed epithelial cell nuclei (DAPI) with a normal branched Y-suture (F-actin) under the epithelial cells. In contrast, there is an obvious defect at the apex of the 18-month-old lens with abnormal distribution of epithelial cell nuclei (DAPI, arrows) and a gap in the anterior suture (F-actin, arrowheads). (B) A 2D YZ projection of the 3D reconstruction of a Z-stack through the anterior epithelium and underlying fiber cells in the 4-month-old lens reveals tight adhesion of the anterior epithelial and fibers. In the 18-month-old lens near the fiber cell defect, the anterior epithelial cell layer is wrinkled and is depressed into the gap of the Y-suture. Although there was a defect in the epithelial cell sheet organization, there was no evidence of multilayered epithelial cells or abnormal epithelial cell proliferation in the 18-month-old lens. These results reveal that anterior cataracts in 18-month-old lenses are correlated with detachment and wrinkling of the anterior epithelial cells from the underlying Y-suture formed by fiber cells. Scale bars, 0.5mm in A and 20μm in B.