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Fig. 4. Expression of dnRobo2-MT in developing RGCs reduces dendrite outgrowth and branching. Examples of transgene-expressing RGCs in sections of stage 40 Xenopus retinas following electroporation at stage 27. Cell morphology was analyzed for RGCs expressing GFP alone (A), GFP plus dnRobo2-MT (B), or GFP plus dnRobo3-MT (C). (D–F) anti-myc immunoreactivity (E) for dnRobo2 accurately reflects the dendritic arbour as revealed by GFP fluorescence (D). Merge is shown in F. (G–L) dnRobo2-expressing cells in the RGC layer (G, J) express differentiated protein markers of RGCs, including the transcription factors islet-1 (H, I) and Brn3b (K, L). (M–O) A dnRobo3-expressing RGC (M) expresses Pax6 protein (N, O). In all panels the cells are oriented so that the IPL would be at the top and the lens the bottom. Arrows point to primary dendrites extended by the RGCs. Note that dnRobo2-MT and dnRobo3-MT were expressed at lower transfection efficiencies than GFP. Scale bar is 5 μm. |
