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Larger Image Figure 2. EFTF-expressing pluripotent cells form eye-like structures in tadpole flank. (A) Schematic illustrating the expected retinal layers of the eye including the outer nuclear (ONL), inner nuclear (INL), and ganglion cell (GCL) layers. The mitotically active CMZ is indicated in red. (B) Transverse section of a stage 41 wild-type retina. Dashed lines mark the location of the plexiform layers. The eye is oriented so that dorsal is to the left. (C) Flank eyes expressed YFP throughout, contained an RPE and a trilayered morphology with putative inner and outer plexiform layers (dashed lines in D). Markers were used to detect retinal cell classes. (C) Whole tadpole bright field image overlayed with fluorescent image, showing flank eye (yellow box) on top of gut (dashed line). Inlay is a magnified view of the boxed flank eye. (D) Cryostat section (12 ) shows cells labeled for the retinal ganglion cell (RGC) marker, hermes, by in situ hybridization. (E) R5 antibody labels Mler glia (M red), which extend processes the breadth of the flank retina. (F) GABA labels horizontal and amacrine cells (HC, AM; red) of the INL. (G) Cells expressing YFP (green) co-express the cone photoreceptor (cPR) marker Calbindin (yellow). (H) Cells expressing the rod photoreceptor marker XAP2 (rPR; red); BrdU-immunoreactivity (yellow) identifies mitotically active cells in the periphery of the same flank retina. Scale bars, 25 . Image published in: Viczian AS et al. (2009) Viczian et al. Creative Commons Attribution license Permanent Image Page Printer Friendly View XB-IMG-81057 |