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Figure 2. The Translation of the Xenopus Homeobox Xotx5b, Xvsx1, and Xotx2 mRNAs Parallels the Generation of Photoreceptors and Bipolar CellsIn situ hybridization of Xotx2, Xotx5b, and Xvsx1 mRNAs (Fast Red detection) compared to immunostaining of the corresponding proteins (green detection) on serial 10-μm sections of embryonic retinas at st. 34 (mid-neurogenesis), st. 37 (late-neurogenesis), and st. 42 (mature embryonic retina). Schematics show the retinal cell types present at the corresponding times of analysis (see also Figure S2). Dashed lines border the entire thickness of neural retinas (st. 34–37), or indicate the boundaries between different cell layers (st. 42, magnification of central retinal aspect); GCL: ganglion cell layer, INL: inner nuclear layer, ONL: outer nuclear layer. |
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Figure S1. Xotx5b, Xvsx1, and Xotx2 Expression in the CMZ
Comparison between Xotx5b, Xvsx1, and Xotx2 mRNA or protein detection (red) and BrdU-positive cells (green) in the CMZ of st. 42 retinas, after an 8-h BrdU pulse. Since this time of incorporation corresponds to the average cell cycle length of a late embryonic retinal progenitor (see Figure S5), the region of green-labeled cells reasonably excludes the majority of post-mitotic cells. From its most marginal aspect (M) towards the central side of the retina (C), the CMZ recapitulates the different stages of embryonic retinal neurogenesis [23], with more marginal cells earlier (less mature) than more central ones. Green arrows point at the central boundary of BrdU immunodetection, and red arrows indicate the marginal border of mRNA/protein detection. Whereas mRNA detection of the three genes always largely co-localizes with BrdU-labeled cells, none of the cells expressing detectable levels of the corresponding protein contains BrdU. These data indicate that Xotx5b, Xvsx1, and Xotx2 start to be translated at a measurable level in post-mitotic cells.
https://doi.org/10.1371/journal.pbio.0040272.sg001 |
