Figure 2

Visualization of the development of the retinas targeted at E13. (A-I) Retinas from E13 embryos were electroporated with GFP-bearing plasmids and sacrificed at E14, E16 or E18. Left panels show retinal sections from electroporated embryos incubated with anti-Islet1/2 antibodies to detect post-mitotic RGCs. Middle panels show targeted cells in the same retinal sections. Note that axons projecting to the inner layer can already be visualized in panel B at E14. Right panels show the merged images. At E16 GFP-positive cells are located closer to the inner layer (labelled by Islet 1/2, red) and a few double-labelled cells are observed (white arrows). At E18 the majority of the electroporated cells are located in the inner retinal layer and many of them are positive for Islet1/2. Scale bar: 20 μm. (J) Diagram showing the retrograde labelling paradigm. Dextran-rhodamine is applied at E17 in the optic tract (red) contralateral to the retina that was electroporated at E13 (green). The typical distribution of dextran-labelled cells and axons in the contralateral retina at E17 are shown (red), together with the GFP targeted cells that were electroporated at E13. (K) Retinal section electroporated at E13 (green cells) and retrogradely labelled with dextran-rhodamine (red cells). In all of the merged images, the double/labelled cells are yellow and they are indicated by white arrows. Scale bar: 100 μm (L-N) High magnification of the boxed area in K. Scale bar: 25 μm INL, Inner layer; VZ, ventricular zone.