Fig. 2: Brief exposure to P.e. infection or DSS triggers ISC migration towards sites of injury. | Nature Communications

Fig. 2: Brief exposure to P.e. infection or DSS triggers ISC migration towards sites of injury.

From: Pvf1-Pvr-mediated crosstalk between trachea and gut guides intestinal stem cell migration to promote gut regeneration

Fig. 2

a–d Guts expressing E-Cad::GFP and Moesin::GFP from endogenous promoters were used to visualize adherens junctions (AJs) and the apical epithelial surface in control (a–a’), DSS-treated (b–b’), and P.e.-infected (c–c’) flies. In controls, ingested fluorescent dextran (red) remains luminal. DSS and P.e. cause epithelial disruption indicated by loss of E-Cad::GFP/Moesin::GFP signals and dextran leakage (white arrows in (b–c’), n = 16, 14, 14 guts). e–f”’ Guts with E-Cad::GFP (white) and ISCs (cytoplasmic white, ISCts > YFP) after 1 h DSS feeding. Loss of membrane E-Cad was used as a proxy for epithelial damage. Live imaging over 6 h revealed ISCs migrating towards an area with disrupted E-Cad “pattern” (orange stippled circle, e–e”’, g, h, n = 5/11, 5/19 guts/ISCs). In intact E-Cad areas (green stippled circle), ISCs remained static (f–h). i–i”’ Tracked trajectories following a 2 h P.e. infection show directed ISC (white) migration toward a damaged region, identified by the loss of E-Cad-GFP signal and dextran leakage. j–n Guts with EC-specific expression of the apoptotic reporter, Apoliner, infected for 2 (k) or 16 (l) hours to induce localized or widespread damage. To induce Apoliner/fluorophore expression in all ECs, mexts>Apoliner flies were shifted to the permissive temperature (29 °C) for 24 h prior to infection, then returned to room temperature (RT, non-permissive) and infected with PE for 2 h (OD200) or overnight (OD100). As Gal80ts is reactivated at RT, newly regenerated ECs are marked by the loss of fluorophore signal (black areas, stippled lines). A 2-h infection led to small, localized renewal zones (k, m, n), while a 16-h infection caused widespread renewal (l–n; m: n = 5/26, 12/189, 13/46 guts/renewed regions, n: n = 5, 11, 13 guts). o–o”’ Optogenetics damage was induced by expressing a light-activatable OptoDronc in small EC clones. ISC trajectories show directed movement towards apoptotic ECs (marked by yellow stars). Significance was tested with a two-tailed Mann–Whitney test (g, h), or Kruskal–Wallis with post-hoc multiple comparison analysis (for the rest). Data are presented as mean values ± SD. Source data are provided as a Source data file.

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