Figure 8

Barhl1a:GFP in the preoptic diencephalon marks populations of mitotic progenitors and differentiating neurons. Images represent single confocal Z-stacks through the diencephalon of Tg(barhl1a:GFP;atoh7:gap43-RFP) embryos at 35 hpf (A, A’) and 30 hpf (B, C). Frontal view, dorsal is up. Embryos in (A-B’) have been immunohistochemically stained with anti-GFP, anti-HuC (A, A’) and anti-pH3 (B, B’) antibodies (in red) as well as counterstained with DAPI (in blue). (A’) magnifications of the images in (A) in correspondence with the barhl1a:GFP expressing cell bodies. (A) yellow arrowheads point at the optic nerves from each side marked by Atoh7:gap43-RFP (in gray). White arrows point at Barhl1a:GFP cells co-labelled with HuC, a marker of differentiating neurons located in the basal half of the neuroepithelium. In (B) and its magnification (B’), Barhl1a:GFP cells are also co-labelled by pH3, which marks mitotic neurons located at the apical surface of the neuroepithelium (white arrowhead in B’). (C) 30 hpf Tg(barhl1a:GFP) embryo injected with the gfap:lssKate DNA construct shows sparse LssKate labelling in the diencephalon (in red); which co-localize with GFP (green). Scale bars: (A–C) 50 µm, (A’,B’) 20 µm.