mouse model wild type mice  (Jackson Laboratory)

Journal: Journal of Inflammation Research

Article Title: Nociceptor Neurons are Involved in the Host Response to Escherichia coli Urinary Tract Infections

doi: 10.2147/jir.s356960

Figure Lengend Snippet: Figure 1 CFT073 induces neuron activation and CGRP release (A) Representative Fura-2 ratiometric fields (left) and calcium traces (right) of DRG neurons responding to CFT073 (5 x 107 CFUs), LPS (10 ng/mL) and capsaicin (1 μM) (n=3). (B) Experimental schematic of TRPV1+ neuron ablation and CFT073-induced UTIs to mice. (C) Representative images of TRPV1+ (red) neurons by immunofluorescence assays (n=5). (D) Measurement of CGRP release with an ELISA kit. CGRP released from bladder (0, 8 and 24 hours) after CFT073 infection (5 x 107 CFUs) of vehicle-treated or Caph mice (n = 3; ns, P > 0.05; ***P < 0.001).

Article Snippet: Bacterial Strain, Culture and Mouse UTI Model Uropathogenic wild-type E. coli strain CFT073 (American type culture collection, 700,928) was grown at 37 °C with 5% CO2 in Luria-Bertani broth for 24 hours and resuspended in PBS to an optical density at 600 nm of 0.6.

Techniques: Activation Assay, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Infection

Journal: Journal of Inflammation Research

Article Title: Nociceptor Neurons are Involved in the Host Response to Escherichia coli Urinary Tract Infections

doi: 10.2147/jir.s356960

Figure Lengend Snippet: Figure 2 Nociceptor neurons mediate bladder epithelial function and bacterial load during UPEC infection Vehicle-treated mice were randomly divided into uninfected or infected group. Caph mice were infected equivalent CFT073 (5 x 107 CFUs). (A) H&E Staining of bladders from groups “vehicle-uninfected”, “vehicle-infected” and “Caph- infected” (n=6). (B) Representative images of WGA-FITC in superficial bladder epithelial cells (n=4). (C) Representative images of trypan blue staining in bladders (n=4). (D) Bacterial load recovery (log10 CFU) from urine in “Caph-infected” or “vehicle-infected” group (n =22-24; **P < 0.01). (E) Ex vivo gentamicin protection assays representing invaded bacteria in “Caph-infected” or “vehicle-infected” group (n =12; **P < 0.01).

Article Snippet: Bacterial Strain, Culture and Mouse UTI Model Uropathogenic wild-type E. coli strain CFT073 (American type culture collection, 700,928) was grown at 37 °C with 5% CO2 in Luria-Bertani broth for 24 hours and resuspended in PBS to an optical density at 600 nm of 0.6.

Techniques: Infection, Staining, Ex Vivo, Bacteria

Journal: Journal of Inflammation Research

Article Title: Nociceptor Neurons are Involved in the Host Response to Escherichia coli Urinary Tract Infections

doi: 10.2147/jir.s356960

Figure Lengend Snippet: Figure 3 Nociceptor neurons suppress recruitment and bactericidal function of neutrophils (A) Representative FACS plots (left) showing neutrophils (CD11b+Ly6G+ gates) in Caph or vehicle-treated mouse bladders. Assessment ratio (right) of Caph or vehicle-treated mice bladder neutrophils by flow cytometry analysis (n=3; *P < 0.05). Mouse neutrophils were co-cultured with CFT073 in presence of vehicle or CGRP (1μM) for 1 hour (B–D). (B) Myeloperoxidase (MPO) activity of neutrophils (n=5; ***P < 0.001; #P < 0.0001). (C) Bacterial phagocytosis of neutrophils (n=6; ns, P > 0.05). (D) Bacterial killing function of neutrophils (n=6; *P < 0.05).

Article Snippet: Bacterial Strain, Culture and Mouse UTI Model Uropathogenic wild-type E. coli strain CFT073 (American type culture collection, 700,928) was grown at 37 °C with 5% CO2 in Luria-Bertani broth for 24 hours and resuspended in PBS to an optical density at 600 nm of 0.6.

Techniques: Flow Cytometry, Cell Culture, Activity Assay