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Fig. 4. 
A-crystallin expression prevents cataract formation in
cloche. A- (panel 3) or 
-crystallin (panel
4) mRNA was injected into embryos at the one- to four-cell stage and
phenotypes were compared with uninjected wild type (panel 1) and
cloche lens (panel 2). (A) Living embryos were analyzed by
stereomicroscopy and the number of opaque lenses was counted. Red arrows point
to scattered light. The percent of lenses that demonstrate opacity is shown in
parentheses. (B) Living embryos lenses were analyzed by confocal
microscopy and the intensity of reflected light was measured. Red arrows
indicate areas of reflected light. The median reflectance was measured as in
Fig. 3D. The Mann-Whitney U test was used to test for differences
between two groups. The differences in the lens between cloche
embryos and cloche embryos overexpressing
A-crystallin (P=0.0433) and the differences between
cloche overexpressing A-crystallin versus
-crystallin (P=0.0209) are significant. (C)
Sections were prepared from 4 dpf embryos and analyzed by Hematoxylin and
Eosin staining. The green arrows indicate nuclei. The number of lens fiber
cell nuclei per section was analyzed at 4 dpf. Three sections of each lens
were analyzed by an image analysis program (ImageJ). Fourteen embryos were
analyzed. At 4 dpf, wild type and cloche had 0.0±0.0 and
22.6±5.0 nuclei, respectively. In the rescue experiment,
A-crystallin overexpression reduced the nuclei from
22.6±5.0 to 3.0±0.0 (mean±s.d.).
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