Immunology and Infection
Published: December 1st, 2017
The established model of transurethral catheterization of mice allows the study of bladder pathologies, including urinary tract infection, but can only be performed in females. A new model of male transurethral instillation, presented here, will enable research in an area marked by strong clinical and epidemiological differences between the sexes.
Urinary tract infections (UTI) are extremely common worldwide, incurring significant morbidity and healthcare-associated expenses. Small animal models, which accurately reflect disease establishment and progression, permit dissection of host-pathogen interactions and generation of immunity to infection. In mice, intravesical instillation of uropathogenic E. coli, the causative agent in more than 85% of community acquired UTI, recapitulates many of the stages of infection observed in humans. Until recently, however, UTI could only be modeled in female animals. This limitation has hindered the study of sex-related differences in UTI, as well as other bladder pathologies, such as cancer. Here, we describe a method to instill male mice that allows direct comparison between female and male animals and provide a detailed protocol to assess bladder tissue by flow cytometry as a means to better understand host responses to infection. Together, these approaches will aid in the identification of host factors that contribute to sex biases observed in UTI and other bladder-associated diseases.
Urinary tract infections (UTI) are one of the most common infections in developed countries1. Infection rates are similar between females and males among neonates and the elderly2. Premenopausal adult women, however, have a greatly increased incidence of community-acquired UTI compared to men2,3. Given that this disease primarily impacts women, fundamental and clinical research has overwhelmingly focused on UTI in females. However, UTI in men is a significant and understudied health care challenge4. Indeed, because UTI in men are associate....
Mouse experiments were conducted in accordance with approval of protocol number 2012-0024 by the Comité d'éthique en expérimentation animale Paris Centre et Sud (the ethics committee for animal experimentation), in application of the European Directive 2010/63 EU.
1. Preparation of Catheters
In the development of this protocol, cohorts of female and male C57Bl/6 mice aged 6 to 8 weeks were instilled, and bacterial burden evaluated in bladders at time points ranging from 1 hr to 30 days. The results from these studies are detailed elsewhere (Zychlinsky Scharff et al., pending). Here, we present representative data from 24 hr infections. Notably, bacterial burden was equivalent between male and female mice at 24 hr post-infection (Figure 1). Little var.......
Transurethral instillation of male mice offers many new research opportunities into the influence of sex on bladder mucosal disease, but also presents several challenges. Foremost, one limitation is that the instillation may initially prove to be technically difficult, resulting in excessive inflammation during catheter insertion. Improvement in technique can be achieved by the instillation of dead mice with a colored solution, such as Evans blue dye. To confirm that the instillation is successful, bladders should be vis.......
We thank Dr Matthieu Rousseau for critical reading of the manuscript and the Laboratory of Dendritic Immunobiology for their helpful insights during the development of this protocol and project. This work was supported in part by funding from the European Union Seventh Framework Programme Marie Curie Action (PCIG11-GA- 2012-3221170, and the Immuno-Oncology LabEx (MAI).....
|BD Insyte Autoguard Shielded IV Catheters 24G, 0.7 mm external diameter, 14 mM long
|381811 or 381411*
|* catalog number is country-dependent
|LB Miller broth
|LB Miller agar
|Thumb (Adson) forceps
|Fine Science Tools
|5 mL tubes, polypropylene
|Tissue Ruptor disposable probes
|15 mL flip cap tubes
|should be aliquotted and stored at -20 °C to avoid repeated freeze-thaw
|100 µM MACS SmartStrainers
|compatible with 15 mL tubes, preferred
|100 µm cell strainers
|compatible with 50 mL tubes, if MACS Smart Strainers are not available
|96 well plate
|anti-mouse CD45 antibody
|many different fluorescent conjugation options are available
|5 mL tubes polystyrene with filter cap
|hand held homogenizer
|Flow cytometry software
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