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. 2011 Apr 18;121(5):1974–1985. doi: 10.1172/JCI44900

Figure 3. Neuron-derived Sema3E restricts vascular pathfinding in developing retinas.

Figure 3

(A) ISH in P4 retinal sections. Plxnd1 is expressed in vascular ECs and in 7.4% of the neuronal populations in the GCL. Sema3e is expressed in 37% of neurons in the GCL. INL, inner nuclear layer. (B) High-magnification image of the advancing vascular front in AP-stained retina after intravitreal injection of Sema3E-AP fusion protein. (C) Whole-mount PECAM-1 IHC in P4 retinas. In the Sema3e–/– retina, the vascular network is disorganized in the upper-right quadrant. Note the attachment of hyaloid vessels to the retinal surface (arrowhead). (D) Quantification of filopodium length (n = 18 per group) and filopodium number/100 μm vessel length (n = 7 per group) at the sprouting vascular fronts, and vascular density (n = 4 per group) in P4 Sema3e-deficient retinas. Error bars represent SEM; ***P < 0.001, *P < 0.05. (E) Triple labeling for neurofilaments (green), PECAM-1 (red), and nuclei (blue) in retinal sections of P4 Sema3e-deficient mice. The superficial vessels (arrowheads) over the neuronal axons ectopically invade the GCL of Sema3e–/– mice (arrow). (F) A schematic diagram representing VEGF-expressing astrocyte (AC, red), PlexinD1-expressing EC (green), and Sema3E-expressing neuron (blue) in the superficial area of the developing mouse retina. Scale bars: 20 μm (A, B, and E); 100 μm (C).