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Figure 3. Correlation of the expression patterns of Notch pathway components with that of a stem/progenitor marker in the metamorphosing intestine. Cross-sections of the intestine from tadpoles at stages 60 (A, D) and 61 (B, E, G, I), and stage-54 tadpoles treated with 10 nM T3 for 3 (C, F) and 5 days (H, J) were hybridized with antisense Hairy1 (A-C) or Hairy2b (G, H) probes. The corresponding serial sections were hybridized with antisense LGR5 probe (D-F, I, J) for comparison. Hairy1 is expressed in the adult epithelial stem (AE)/progenitor cells (A, arrowhead) when they first appear as the small islets expressing LGR5 (D, arrow) at stage 60. These islets grew in size as metamorphosis proceeded and continued to co-express Hairy1 (B, arrowheads) and LGR5 (E, arrows). During T3-induced metamorphosis, Hairy1 (C, arrowhead) is also expressed in the AE/progenitor cells expressing LGR5 (F, arrow) after 3 days of T3 treatment. Some, although not all, cells expressing Hairy2b were in the connective tissue (CT) (G, arrowheads) underlying the AE/progenitor cells expressing LGR5 (I, arrows). Hairy2b is also expressed in the CT after 5 days of T3 treatment (H, arrowheads), again, with some underlying the AE/progenitor cells (J, arrows). Cross-sections of the intestine from tadpoles at stage 62 were double-immunostained with anti-CK19 to detect the islet of AE/progenitor cells (K-M, red) and anti-DLL1 (K, green), anti-Jag1 (L, green), or anti-Notch1 (M, green) followed by counterstaining with DAPI (K-M, blue). DLL1 (K, arrowheads) and Notch1 (M, arrowheads) are expressed in the islets (arrows). The cells expressing Jag1 are scattered in both the larval Ep and CT with some just beneath the islet, but not in the islets (L, arrowheads). The dashed-lines indicate the boundary of the Ep and the CT. Scale bars = 20 μm. Abbreviations: AE, adult epithelial stem/progenitor cells, CT, connective tissue, Ep, epithelium. Image published in: Hasebe T et al. (2017) © 2016 The Authors STEM CELLS published by Wiley Periodicals, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives license Larger Image Printer Friendly View

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