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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Nature. 2015 Oct 28;527(7578):371–374. doi: 10.1038/nature15758

Figure 3. Bipolar tail neuron specification and differentiation.

Figure 3

b, Wild-type Neurogenin (Neurog) b-line>unc-76∷VenusYFP expression (green) in embryos treated with DMSO vehicle, counterstained with DAPI (blue). b, Neurog expression was abolished in 43/50 embryos treated with 10 μM MEK inhibitor U0126 at 5.5 hpf. c, Supernumerary bipolar tail neurons (BTNs) were specified in 28/50 embryos treated with 10 μM U0126 at 7 hpf. d, Two BTN precursors migrating on one side of a DMSO-treated embryo. e, Expanded chain of four BTNs resulting from treatment with U0126 at 7 hpf. f, BTN expressing Asic>unc-76∷eGFP reporter in embryo electroporated with Neurog b-line>nls∷lacZ as a control. g, Overexpression of Neurog induces specification of ectopic Asic+ BTNs in 53/100 embryos. h, Overexpression of a dominant repressor form of Neurog (Neurog∷WRPW) abolishes BTNs in 97/100 embryos. i, In situ hybridization reveals expression of Cadherin.b (green) in the neural tube but not migrating BTN precursors, and j, expression of Protocadherin.c (green) in dorsal epidermis midline but not BTNs. Embryos in i,j electroporated with Neurog b-line>unc-76∷mCherry (immunolabeling of mCherry in magenta). k, Forced overexpression of Protocadherin.c in the BTN lineage inhibits delamination and migration of BTNs in 7/14 embryos. l, Normal BTNs as seen in 9/12 control embryos (overexpression of β-galactosidase instead). Embryos in k,l electroporated with Neurog b-line>unc-76∷VenusYFP and Neurog b-line>H2B∷VenusYFP (green) and counterstained with phalloidin (magenta). All scale bars, 25 μm. Embryos in a-e, i, j fixed at stage 22. Embryos in f-h, k,l at stage 23.