Fig. 10

Cell-autonomous effect of cilia reduction on RGC differentiation. a-b Blastomeres from atoh7:gap-RFP embryos injected with control MO (12 ng) (a) or elipsa-SP/ift88-SP MOs (5/5 ng) (b) were transplanted into atoh7:gap-GFP hosts. The dotted line indicates de apical margin of the RGC layer of the host and the asterisk denotes the donor cells with high RFP expression. c-d Blastomeres from un-injected atoh7:gap-GFP embryos were transplanted into atoh7:gap-RFP hosts injected with elipsa-SP/ift88-SP MOs (6/6 ng). Retinas with small (c) and large (d) clones of GFP positive cells were observed. In C the dotted line indicates de apical margin of the RGC layer formed by the donor GFP positive cells, and the asterisk denotes host cells with high RFP expression. Single arrowheads mark GFP expressing cells that appear before RFP expressing cells (double arrowheads). In all the cases (a-d) the resulting host embryos were imaged by time-lapse confocal microscopy beginning at around 30 hpf. Fluorescence images show montages from 3D maximum intensity projections of the stacks from three different time points. On the left, the external phenotype at 48 hpf of the donor and host embryos is shown in each case. Time is expressed in hrs:min. Scale bars: A-D, 20 μm. e Comparison of the number of mitotic cells (identified by anti-pHistone H3 labeling) in the ventro-nasal region of 24 and 36 hpf retinas, expressed as the number of positive cells per unit of volume. (*) p = 0.0014. f Comparison of the number of apoptotic cells (identified by anti-activated Caspase 3 labeling) per retina in 36 and 48 hpf embryos. (*) p = 0.009. All experiments were performed twice, with similar results; only the results from one of the experiments are shown. The numbers in brackets represent the number of embryos quantified in each case (we quantified one eye per embryo). (ns) non-significant difference. Comparisons were made using rank transformation and Student’s t test. For a high resolution image of Fig. 10 please see Additional file 21