Skip to main content
. 2022 Jan 11;7(1):e152676. doi: 10.1172/jci.insight.152676

Figure 3. Time-lapse in vivo MPM imaging of the effects of hyaluronidase and ACEi treatment in late-stage AS mice.

Figure 3

(A–H) Acute hyaluronidase (H) treatment in control (A and C) and late-stage AS mice (B and D). Plasma was labeled with albumin–Alexa Fluor 680 (gray), glycocalyx with FITC-WGA lectin (green; linear pattern), and immune cells with anti-CD44–Alexa Fluor 488 antibodies (green; round cell pattern). (A–D) Projection 5-minute time-lapse images (overlay of plasma albumin, gray; glycocalyx and immune cell labeling, green; tissue autofluorescence, orange; insets show glycocalyx and CD44+ cells separately) of the glomerular microenvironment before (A and B) and within 1 hour of i.v. H (50 U) injection (C and D) in control (A and C) and late-stage AS (B and D). Note the high glomerular capillary albumin intensity in late-stage AS mice and its reductions after H treatment (B and D), indicating improved blood flow (flow of nonfluorescent red blood cells [RBC] rather than only the highly fluorescent plasma; Supplemental Video 1). (E–H) Summary of the effects of H treatment in control and late-stage AS mice on GEC glycocalyx thickness (E), CD44+ cell number per glomerulus (F), glomerular capillary blood flow (RBC velocity; G), and glomerular albumin leakage (albumin GSC; H). (I–O) Chronic treatment with H, ACEi, or control PBS for 1 week in late-stage AS mice. Projection 5-minute time-lapse images, as in A–D (insets show glycocalyx (green) and CD44+ cells (red) separately), in control vehicle (PBS) (I), H (J), and ACEi treatment groups (K). (L–O) Summary of the effects of control PBS, H, or ACEi treatment on GEC glycocalyx thickness (L), CD44+ cell number per glomerulus (M), glomerular capillary blood flow (N), and albuminuria (albumin/creatinine ratio [ACR]; O). Data are shown as the mean ± SEM. **P < 0.01, ***P < 0.001, ****P < 0.0001, using 1-way ANOVA followed by Tukey’s or Šidák’s multiple-comparison test. Scale bar: 50 μm. Data points represent the average of multiple measurements/mouse (O) or from 2 glomeruli/mouse (E–N); n = 5–6 mice in each group.