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View full-text article in PMC

. 2018 Dec 27;7(3):655–678. doi: 10.1016/j.jcmgh.2018.12.007

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© 2018 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Histopathologic analysis of moderate and marked HSCR/HAEC in P18–P25 Gfra1^hypo/hypomice. (A–C) Exacerbated mucin type changes, epithelial damage, scarce inflammation in the lamina propria, and a lack of bacterial invasion in P18–P25 Gfra1^hypo/hypo mice with moderate-grade HAEC. (A) In WT mice, the crypts in the colon (A1 and A2, white arrow) are relatively shallow and straight, and the surface epithelium is even and regularly palisading (A1, curved black arrow). The goblet cells in the surface epithelium and in the proximal parts of the crypts contain PAS-positive (ie, neutral) mucins, staining deep purple (A2, white arrowhead). In the basal parts of the crypts, apical mucin droplets in the colonocytes and the mucus in the goblet cells stain primarily with AB, staining light blue (mucin droplets; A2, yellow arrowhead) to dark blue (goblet cells; A2, open arrowhead). In A3, gram-positive (blue to black) and gram-negative (intense red) bacteria are isolated from the surface epithelium by a layer of mucus (A3, black arrow). (B) The intestinal lumens of Gfra1^hypo/hypo mice contain fecal material and copious amounts of mucus with eosinophilic debris of shed epithelial cells (B1–B3, white bold arrows). The crypts were markedly dilated with retained mucin (B1–B3, white arrow). The surface epithelium had diffuse degeneration (B1 and B2, curved black arrow) and an increased number of goblet cells (B1–B3, white arrowhead). Multifocal necrotic and apoptotic colonocytes (B2 and B3, open white arrowhead) and scattered intraepithelial neutrophils (B2, red open arrowhead). (C) The basal lamina propria contains moderately increased numbers of eosinophils (C1, black arrowhead) and mildly increased numbers of lymphocytes (C1, yellow open arrowhead), indicative of an inflammatory process. Note the reduction in AB-stained (ie, acidic) mucin (C2, black open arrowhead) in the cytoplasm of basal crypt colonocytes. Gram staining shows copious amounts of mucus (C3, black bold arrow) and a lack of bacteria in mildly dilated crypts (C3, white arrow). (D and E) Marked epithelial damage at the surface epithelium, crypt abscesses, bacterial invasion, and inflammation in P18–P25 Gfra1^hypo/hypo mice with marked HAEC score. (D) In Gfra1^hypo/hypo mice, crypt dilatation (D1, white arrow) along with sparse neutrophil infiltrates in basal (D1, red open arrowheads) and apical (D2, red open arrowheads) crypt epithelium. The superficial epithelium shows multifocal necrotic areas and degenerative changes accompanied by superficial dense infiltrates of neutrophils (ie, microabscesses; D2, green arrowheads). Crypt abscesses (E1, green arrow), dense bacterial aggregates (D3, black arrow), and enterocyte-attaching bacteria (D3 and E2, green bold arrow) also are indicated. Bacteria inside a submucosal blood vessel (E3, black arrow) and a bacterial aggregate in mildly dilated crypt base (E3, white arrow) are present. Single granulocytes (E3, yellow bold arrows) and lymphocytes (E3, yellow arrowheads) are present in the lamina propria and submucosa, indicative of inflammation.