Fig. 5: Enteric nitrergic neuron and glial cell dynamics in chronic T. cruzi infections and after benznidazole treatment. | Nature Communications

Fig. 5: Enteric nitrergic neuron and glial cell dynamics in chronic T. cruzi infections and after benznidazole treatment.

From: Enteric nervous system regeneration and functional cure of experimental digestive Chagas disease with trypanocidal chemotherapy

Fig. 5

a Representative immunofluorescent confocal z-stack whole-mount images of nNOS+ neurons in the myenteric plexus of the mouse colon. b Bar plots show number of nNOS+ neuronal cell bodies per field of view in control (n = 10), infected (n = 9), benznidazole treated and cured (BZ-Cured, n = 6 in proximal [PC] and n = 7 in distal colon [DC]), and benznidazole treated and relapsed (BZ-Relapsed, n = 9 in PC and n = 8 in DC) C3H/HeN mouse proximal and distal colon regions. c Representative immunofluorescent confocal z-stack images to display changes in anti-GFAP (gold yellow colour intensity scale) stained enteric glial cells (EGCs) co-labelled with anti-TuJ1 (blue colour intensity scale) enteric neural network across different experimental groups in the colon myenteric plexus. Top panel shows merged images of GFAP and TuJ1 labelled cells. Bottom panel shows images of morphologically diverse GFAP+ EGCs (red pixel colour intensity scale) in the myenteric plexus of infected and BZ-Relapsed colons compared to control and BZ-Cured. White arrows show indicate putative degraded GFAP+ EGC and white stars indicate activated GFAP+ EGC morphologies. All confocal images (a, c) were taken at 400× magnification, scale bar = 50 µm. Colour heat map scales show pixel intensity. All micrographs are representative images of two independent experiments. d Bar plots and e paired dot plot show Western blot analysis of GFAP protein abundance in whole tissue lysates from mouse colons (as a ratio of control levels) from infected, BZ-Cured and BZ-Relapsed (n = 3, biological samples, all groups). Representative immunoblot in d shows α-GFAP staining using 12 µg of lysate (corresponding to the bar plot groups above). To demonstrate equal sample loading, the most abundant protein in each group is presented below as a stain-free gel image. For comparison, GFAP abundance quantified by Western blotting of 12 and 21 µg of each lysate is indicated in plot (e).

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