Fig. 5
Pax6b but not NeuroD is sufficient to induce endodermal hb9 expression. (A) Schematic of the induction experiment: One cell of a 16–32 cell embryo stage was injected with mRNA encoding either Cas alone or a mixture of mRNAs encoding Cas together with NeuroD or Pax6b. Injected embryos were fixed 8 h later (9 hpf) and analysed by mRNA in situ hybridisation. (B–E) Expression of hb9 mRNA in uninjected control embryos (B) and in embryos injected with cas (C), cas/paxb6 (D) and cas/neuroD (E) mRNA. Note that only cas/pax6b mRNA injected embryos (D) show ectopic hb9 signals. Signals in posterior notochord (bracket in B, C, E) represent endogenous hb9 expression, while reduced signals in the cas (C), and cas/neuroD (E) injected embryos correlate with the developmental delay caused by the injection. (F–I) Expression of gfp mRNA in transgenic reporter lines in which gfp is controlled by PE-containing (Tg[hb9:GFP]ml2 in F–H) or PE-deleted (Tg[hb9-NheI:GFP] in I) hb9 promoter-sequences. Uninjected (F) and cas-mRNA injected Tg[hb9:GFP]ml2 embryos (G) show no gfp signals while injection of cas/pax6b mRNA results in a robust gfp induction (H). Lack of gfp signals in cas/pax6b injected Tg[hb9-NheI:GFP] shows that gfp induction by Pax6b is PE-dependent. |
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| Stage: | 90%-epiboly |
Reprinted from Developmental Biology, 365(1), Arkhipova, V., Wendik, B., Devos, N., Ek, O., Peers, B., and Meyer, D., Characterization and regulation of the hb9/mnx1 beta-cell progenitor specific enhancer in zebrafish, 290-302, Copyright (2012) with permission from Elsevier. Full text @ Dev. Biol.