αA-crys knockdown with morpholinos (MOs). (A- E) Effects of MO on lens development. (A) Embryos injected with control MO have a normal sized lens. (B- D) Embryos injected with αA-crys splice-blocking morpholinos (sbMO) and translation-blocking morpholino (tbMO) develop a normal sized lens (B), a lens of reduced size and a reduced ventral optic cup (VOC) (C)1, or no lens (D). (E) Embryos injected with αA-crys sbMO and αA-crys mRNA show normal lens development. (F- J) Apoptosis detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). (F) An uninjected embryo treated with DNase shows apoptotic cells throughout the eye. (G, H) Uninjected (G) and control MO-injected (H) embryos show background levels of lens apoptosis. (I) Embryos injected with αA-crys sbMO and tbMO embryos show apoptotic cells in the lens (L) and retina (R). VR: reduced ventral retina. (J) Embryos injected with αA-crys sbMO and αA-crys mRNA show background levels of lens apoptosis. (K) Semiquantitative RT-PCR . SF: uninjected embryos. SF-MO: αA-crys MO-injected embryos. (L, M) In situ hybridization shows mip gene expression in the lens of uninjected (L) and αA-crys sbMO-injected (M) embryos. All embryos are shown at 40 hr post fertilization (hpf). All scale bars are 80 μm; A-E, F-J, L and M are the same magnifications. (N) Histogram showing lens apoptotic cells in control MO-injected embryos (blue), αA-crys sbMO-injected embryos (red), and αA-crys sbMO and αA-crys mRNA-injected embryos (gray). Error bars represent SD. Numbers at the base of the histogram represent sample sizes. P = 0.00 (one-way analysis of variance (ANOVA).