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Fig. 5. Retinal phenotype of 3-month-old Crb1-/- mice exposed to 3000 lux for 72 hours. (A) Representative experiment with wild-type and Crb1-/- retinas indicating number of protrusions, ingressions and total amount in cycled light (12 hours dark/12 hours 100 lux) versus 72 hours of 3000 lux. Error bars represent s.e.m. Asterisks indicate statistical difference (P<0.02) between the groups exposed to cycled light and 72 hours of 3000 lux. (B) Numerous ingression areas through the OPL (arrow) and protrusions through a distorted OLM (arrowhead) in the Crb1-/- retina. (C) OLM present in rosette (arrow), no segments are present on the disorganized PRCs (arrowhead). (D-J) Fluorescence microscopy images of degenerated areas. Nuclei are stained with Hoechst (blue). Note that in unaffected areas, adjacent to the ingression areas, localization of the proteins is normal. GFAP localized near the inner limiting membrane, at the MGC end-feet and in horizontally radiating MGC rootlets in the OPL (data not shown) in wild-type mice exposed to both cycled light and 72 hours of 3000 lux. (E) Detail of D, strong staining of GFAP in the ONL and through OLM. (F) Areas of protrusions (arrowheads) and ingressions (arrows) where Mpp4 is lost at the OLM and OPL as well as mislocalized into the ONL (asterisk). (G) ß-Catenin mislocalization throughout the ONL in ingression areas (arrows). (H) ZO-2 localization perturbed in a protrusion and ingression area. (I,J) Mislocalization and loss of Patj or Mupp1 in affected areas (arrowheads). (K) Apoptotic cells are rarely present in wild-type retinas. (L) Slightly increased apoptosis is apparent in Crb1-/- mice around ingression areas. Scale bars: 50 µm. (M) SLO image (514.5 nm) of a 3-month-old Crb1-/- mouse fundus after exposure for 72 hours to 3000 lux. The multiple dots (arrowhead) indicate areas of rosette formation in the inferior temporal quadrant of the retina. Scale bars: 50 µm.
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