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Fig. 6.
Prdm13 expression is deregulated upon Ptf1a gain of function. Whole-mount in situ hybridization analysis of prdm13 and gad1 expression in ptf1a-GR mRNA injected embryos treated with dexamethasone (Dex) and analysed at the indicated stages. Shown are lateral views of the embryos (a), of the head at higher magnification (b), and transversal sections of the retinas (c). The number of analysed embryos and percentage of embryos with represented phenotype are indicated in each panel. Scale bars represent 400 μm (a, b) or 100 μm (c) |
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Fig. 2.
Prdm13 is expressed in glycinergic- and GABAergic-amacrine cells. a Double in situ hybridization with prdm13 (red) and gad1 (green) probes on stage 39/40 retinal section. bIn situ hybridization with prdm13 probe (red) coupled with anti-Glycine immunostaining (green) on stage 41/42 retinal section. Arrows point to double labelled cells. Dotted lines separate the GCL and the INL. c Quantification of the percentages of GABA (gad1
+), Glycine (Gly) and Calretininin (Cal) amacrine cells among prdm13
+ cells in the INL and in the GCL. Data are presented as mean ± SEM. Number of analysed sections is indicated in each bar. GCL: Ganglion Cell Layer, INL: Inner Nuclear Layer. Scale bar represents 25 μm |
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Fig. 4.
Prdm13 loss of function leads to a decrease in glycinergic but not GABAergic-amacrine cells. a Whole-mount in situ hybridization analysis of prdm13, glyt1, gad1 and vglut1 expression on stage 39/40 embryos injected with prdm13-M0 or control-MO. Lateral views of the head and transversal sections of the retinas are shown. The number of analysed embryos and the percentage of embryos with represented phenotypes are indicated in each panel. b Stage 39/40 sections following GABA or Glycine-immunostaining on control-MO and prdm13-MO injected embryos. Arrows point to Glycine-positive cells. c Quantification of the average number of GABA- or Glycine-positive cells per section. Number of analysed sections is indicated in each bar. Data are presented as mean ± SEM. p < 0.001 (***) (Mann-Whitney test). Scale bars represent 200 μm (whole mount), 100 μm (sections) |
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Fig. 5.
Prdm13 expression in the retina upon Ptf1a gain and loss of function. a Double fluorescent in situ hybridization with prdm13 (red) and ptf1a (green) probes on wild type retina at stage 35/36 and 40. Right panels are enlargement of central or peripheral retina (white squares). Arrows show double labelled cells. b Quantification of the percentage of ptf1a
+ cells among the prdm13
+ cell population (top graph) and the percentage of prdm13
+ cells among the ptf1a
+ cell population (bottom graph) at different stages of retinogenesis. Data are presented as mean ± SEM. Number of analysed sections is indicated in each bar. c Analysis of gad1, prdm13 and glyt1 expression on stage 41 retinal transversal sections following whole mount in situ hybridization on embryos injected with ptf1a-MO, control-MO, GFP mRNA (control) or ptf1a-GR mRNA. Dexamethasone was added at stage 21/22 to activate the Ptf1a-GR fusion protein. Scale bar represents 100 μm |
