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. 2020 Dec 17;11:596457. doi: 10.3389/fphys.2020.596457

Figure 2.

Figure 2

Early feeding increases villus PepT1 expression and does not affect crypt Lgr5 expression at d10. mRNA expression of the stem cell marker Leucin-rich repeat-containing G-protein coupled receptor 5 (Lgr5) detected by RNAscope in situ hybridization (ISH) at Hatch (A, X100 magnification; B, crypt zone by X400 magnification) and d10, in DF (C) and EF (D) chicks (X400 magnification) is restricted to crypts at both timepoints. Probes were detected with fast-red, and tissues were counterstained with 50% hematoxylin. mRNA expression of the enterocyte marker Peptide transporter 1 (PepT1) detected by RNAscope (ISH) at Hatch (E, X100 magnification; F, crypt zone by X400 magnification) and d10, in DF (G) and EF (H) chicks (X100 magnification) is restricted to villi at both timepoints. Probes were stained with 3,3'-diaminobenzidine (DAB), and tissues were counterstained with 50% hematoxylin. Scale bars indicate 50 μm. At d10, Lgr5 probe densities in DF and EF chicks were calculated relative to crypt area (I), and PepT1 probe densities in DF and EF chicks were calculated relative to villus area (J). Values are means + SEM, and asterisks mark significant differences by t-test, p < 0.05.