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. 2014 Feb 13;9:3. doi: 10.1186/1749-8104-9-3

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Copyright © 2014 Bertolesi et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Distinct cell types induce c-fos expression in response to light. (A) Transverse Stage 42 retinal sections co-immunolabeled for Isl1 (green), and either Otx2 (top), Pax6 (middle), or Prox1 (bottom) (red). Higher magnification of a merged picture (right column) and DAPI staining (middle column; blue). The Isl1 antibody recognized On-BCs and ACs in the inner nuclear layer (INL), and RGCs in the RGC layer. Otx2 stained PRs in the outer nuclear layer (ONL) and On-BCs (Otx+/Isl1+) and Off-BCs (Otx+ / Isl1−) cells. Pax6 identified ACs in the INL, and RGCs. Three populations of cells in the RGC layer were defined by Pax6 and Isl1 expression: Isl1+ / Pax6− (RGC1; green), Isl1+ / Pax6+ (RGC2; yellow) and Isl1− / Pax6+ (RGC3; red). Prox1+ HCs were in the outer region of the INL. (B) Light induced c-fos in On-BCs. c-fos mRNA in central retinal sections of dark-reared Stage 42 embryos exposed to light (2500 lux) for 30 minutes, followed by immunohistochemistry against Isl1 (green) or Otx2 (red), and by DAPI staining (blue). A higher magnification of the region is indicated, and the corresponding merges are shown. Two c-fos+ / Otx2+ / Isl1+ (arrowheads) and two c-fos+ / Otx2− / Isl1− (arrows) cells are indicated. The percentage of c-fos + cells in central retinal sections expressing the corresponding markers (mean ± SD; n = 10 retinas) in the INL is shown in tabular form. (C) Two sub-populations of RGCs expressed c-fos. In situ hybridization against c-fos, immunohistochemistry against Isl1 (green) or Pax6 (red), and DAPI staining (blue). Two c-fos+ / Pax6− / Isl1− cells (arrowhead) in the INL, and three c-fos + cells (arrows) corresponding to RGC1 (Isl1+ / Pax6−), RGC2 (Isl1+ / Pax6+), and RGC3 (Isl1− / Pax6+) are indicated. Scale bar = 50 μm for lower, and 10 μm for higher, magnifications.