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Fig. 2. Microphthalmia and disrupted retinal morphology following Notch1 inactivation in the RPCs. (A) Normal eye size is observed in the control, while microphthalmia is evident in the Notch1^f/f;-Cre mice (E). (B-D,F-H) H&E staining of eye sections from P15 (B,F), E17.5 (C,G) and E13.5 (D,H) embryos demonstrating the normal retinal morphology in the control eyes (B-D), while in the Notch1^f/f;-Cre eyes rosettes are detected in the retina (F-H) as early as E13.5 (H). (I) Diagram illustrating the lineage tracing of the Notch1^- cells employing the Notch1^f/f;-Cre;Z/AP mice. In these mice, Notch1 gene remains intact and AP is not expressed in cells that have not expressed Cre. Notch1, however, is deleted and AP activity detected in cells that expressed Cre and in their progeny. (J-Q) AP activity was monitored on sections from control (J-M) and Notch1^f/f;-Cre;Z/AP (N-Q) eyes. At P15 (J,K) and P0 (L), AP is detected throughout the peripheral -Cre;Z/AP retina as expected from the early activity of Cre observed in most RPCs in this region (M). In the Notch1^f/f;-Cre;Z/AP P15 eyes, however, AP⁺ (Notch1^-) cells are dramatically reduced in number thus most of the peripheral retina is missing (N,O). The iris maintains its normal structure despite loss of retinal tissue (compare J with N). At P0 (P), fewer AP⁺ cells are detected in the Notch1^f/f;-Cre;Z/AP retina when compared with the wide distribution of AP⁺ cells in the control (L). The rosettes at P0 are composed of a mixture of AP⁺ (Notch1^-) and AP^- (Notch1⁺) cells. However, at E13.5 (Q), when rosettes first appear in the Notch1^f/f;-Cre;Z/AP, the distribution of AP is similar to its distribution in the controls (M), encompassing most of the peripheral retina and the rosettes seem to be mostly composed of AP⁺ cells (Q, inset). The red arrowheads in J,N,L,P indicate the AP⁺ amacrine precursors that seem to be unaffected following Notch1 inactivation. Control genotypes: Notch1^f/+;-Cre (A-D); Notch1^f/+;-Cre;Z/AP (B-D,J-M). co, cornea; gc, ganglion cells; inl, inner nuclear layer; i, iris; le, lens; nr, neuroretina; onl, outer nuclear layer; rpe, retinal pigmented epithelium. Scale bars: 100 µm in F,H,O,Q; 200 µm in G,P; 500 µm in N.

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