Figure 5.

H. polygyrus infection alters organoid morphology and the intestinal stem cell niche. Developmental changes in the intestinal epithelium were investigated by analysis of organoid morphology and key developmental gene expression following HpES exposure, and expression of the same key genes in intestinal tissue following infection with H. polygyrus. (A) Images 16 h after culture of control organoids and organoids incubated with HpES. Scale bar is 100 µm. (B and C) Quantification of organoid architecture after addition of HpES, showing in the distribution of organoids classed as budding or spheroid in the control (top) and HpES (bottom) treatment conditions (B); and quantification by the area of organoid images over 48 h (C). Unpaired t tests were used for statistical analysis; ****, P < 0.0001. (D–F) Expression of intestinal development–related genes, Atoh1, Neurog3, and Hes1, from organoid cultures under the indicated conditions. Change shown compared with nonstimulated control in four independent biological replicates analyzed in parallel by RNA-seq. One-way ANOVA with Tukey’s multiple comparisons test was used; *, P < 0.05; **, P < 0.01; ***, P < 0.001. (G–I) qRT-PCR on in vivo intestinal samples for the same intestinal development–related genes. Data shown are from five individual mice in one of three replicate experiments. One-way ANOVA with Tukey’s multiple comparisons test was used; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Hp, H. polygyrus; Nb, N. brasiliensis; Stim, stimulation.