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Inhibition of Wnt signaling converts lateral right into lateral left neuronal identities in developing catshark habenulae. a–n Transverse sections of catshark stage 31 habenulae from control (a–g) and IWR-1-treated (h–n) embryos, after IHC with an antibody directed against β-catenin in red and DAPI-staining in blue (a,h), HCR-FISH double-labeling with territories of ScSox1 in yellow (b,i) and those of ScProx1 in cyan (c,j), ISH with probes for ScSox1 (d,k), ScProx1 (e,l), ScNtng2 (f,m) and ScKiss1 (g,n). Sections are shown at a medial organ level, dorsal to the top. (a) and (b,c) show adjacent sections of the same embryo, same for (d) and (e), (f) and (g), (h) and (i,j), (k) and (l), (m) and (n). (a1,a2) and (h1,h2) show magnifications of the territories boxed in (a) and (h), with β-catenin signals in red and DAPI signals in blue. Thin white arrows point to β-catenin labeled nuclei. Arrowheads in (i–n) show expansions of Left-LHb markers to the Right-LHb, in territories where expression of Right-LHb markers and nuclear β-catenin accumulation are lost. “n = “ in (a–n) refers to the number of embryos exhibiting the same phenotype as shown in the figure, over the total number of embryos analyzed, using the same detection method. o Schemes showing the experimental procedure used for pharmacological treatments and the resulting habenular phenotypes, with territories of nuclear β-catenin accumulation in red, territories of Left- and Right-LHb identity in yellow and blue respectively. Abbreviations: LHb, lateral habenula; L, left; R, right, st., stage. Scale bar = 100 µm.
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