A Model of Cystitis Pain in the Mouse

Liza Leventhal1, Brian Strassle2

1 EnVivo Pharmaceuticals, Watertown, Massachusetts, 2 Wyeth Research, Neuroscience Discovery Research, Princeton, New Jersey
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.52
DOI:  10.1002/0471141755.ph0552s43
Online Posting Date:  December, 2008
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Abstract

Interstitial cystitis (IC) is a chronic pelvic-perineal pain syndrome of unknown etiology that mainly targets the lower urinary tract. It is characterized by urinary frequency and urgency, inflammation, stiffening of the bladder wall, and visceral pain. Pain is the most prominent feature of IC and current treatments provide limited relief. To facilitate the identification of novel therapeutic options for IC, efforts are being made to identify more predictive rodent models. A well-characterized rat model is based on administration of cyclophosphamide (CP). The protocol described in this unit replicates this model in mice, demonstrating that CP increases both spontaneous and evoked pain behaviors in this species. This mouse model can be used for understanding disease etiology as well as for the pharmacological evaluation of novel therapeutics for the treatment of IC pain. Curr. Protoc. Pharmacol. 43:5.52.1-5.52.11. © 2008 by John Wiley & Sons, Inc.

Keywords: interstitial cystitis; pain; inflammation; bladder

     
 
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Table of Contents

  • Basic Protocol
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

 Basic Protocol
 Materials
  • Male CD-1 mice (20 to 25 g; Charles River)
  • Cyclophosphamide (CP; Sigma-Aldrich)
  • 0.9% sterile saline (sodium chloride)
  • Test compound
  • Vehicle (control)
  • Behavioral testing apparatus: clear Plexiglas chambers (10.5 × 10.5 × 15.0–cm) on an elevated 21-G perforated steel floor (McNichols, 0.200-in. Hanover Square, ¼-in. straight rows (http://www.mcnichols.com/products/perforated/decorative/stockPatterns.html) (Fig. 5.52.2)
  • von Frey filaments (Semmes-Weinstein monofilaments, Stoelting) with evaluator sizes of 1.65, 2.44, 3.22, 3.61, 3.84, 4.08, 4.17, and 4.31 (0.0008 to 2 g of target force) (Table 5.52.1)
  • 1-ml syringes and 25-G needles
     FigureFigure 5.52.2 A series of testing chambers used for behavioral evaluation. The dimensions of the individual chambers are 10.5 × 10.5 × 15.0–cm. A translucent Plexiglas box is used to allow for visual assessment of the animal behavior. The boxes are placed on an elevated wire mesh floor. The von Frey fibers are applied through the wire mesh floor to the upper abdomen region of the mouse.
     
    Table 5.52.1 Touch-Test Sensory Evaluatorsa
    Evaluator sizeTarget forceColor code
    1.650.008Green
    2.440.04
    3.220.16Blue
    3.610.4
    3.840.6Purple
    4.081
    4.171.4
    4.312
     avon Frey Filaments (Semmes-Weinstein Monofilaments, Stoelting) with specific evaluator size and target force were used for testing withdrawal threshold when applied to the upper abdomen of mice.
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Figures

  •  FigureFigure 5.52.1 Photographic representation of vehicle- (left) and cyclophosphamide (CP, 300 mg/kg, i.p.)-treated (right) mouse bladder 4 hr after CP administration. Note the excessive hemorrhaging and edema in the CP-treated mouse bladder.
    View Image
  •  FigureFigure 5.52.2 A series of testing chambers used for behavioral evaluation. The dimensions of the individual chambers are 10.5 × 10.5 × 15.0–cm. A translucent Plexiglas box is used to allow for visual assessment of the animal behavior. The boxes are placed on an elevated wire mesh floor. The von Frey fibers are applied through the wire mesh floor to the upper abdomen region of the mouse.
    View Image
  •  FigureFigure 5.52.3 Photographic representation of spontaneous pain characteristics following cyclophosphamide (CP) administration: (A) ptosis, (B) licking of the abdomen, and (C) rounded back. (D) Illustration of the rating scale test sheet used during behavioral of spontaneous pain behaviors.
    View Image
  •  FigureFigure 5.52.4 Effect of intraperitoneal cyclophosphamide (CP) administration on spontaneous and evoked pain in CD-1 mice. (A) CP increased spontaneous pain as measured using a rating scale. (B) CP produced referred hyperalgesia as measured by von Frey fibers. Asterisks denote overall significant differences (p < 0.05) from vehicle-treated mice. All data are plotted as mean ± S.E.M.
    View Image

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Literature Cited

Literature Cited
    Bon, K., Lichtensteiger, C.A., Wilson, S.G., and Mogil, J.S. 2003. Characterization of cyclophosphamide cystitis, a model of visceral and referred pain, in the mouse: Species and strain differences. J. Urol. 170:1008-1012.
    Boucher, M., Meen, M., Codron, J.P., Coudore, F., Kemeny, J.L., and Eschalier, A. 2000. Cyclophosphamide-induced cystitis in freely-moving conscious rats: Behavioral approach to a new model of visceral pain. J. Urol. 164:203-208.
    Chaplan, S.R., Bach, F.W., Pogrel, J.W., Chung, J.M., and Yaksh, T.L. 1994. Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53:55-63.
    Cox, P.J. 1979. Cyclophosphamide cystitis—Identification of acrolein as the causative agent. Biochem. Pharmacol. 28:2045-2049.
    Dixon, W.J. 1980. Efficient analysis of experimental observations. Ann. Rev. Pharmacol. Toxicol. 20:441-462.
    Laird, J.M., Martinez-Caro, L., Garcia-Nicas, E., and Cervero, F. 2001. A new model of visceral pain and referred hyperalgesia in the mouse. Pain 92:335-342.
    Laird, J.M., Souslova, V., Wood, J.N., and Cervero, F. 2002. Deficits in visceral pain and referred hyperalgesia in Nav1.8 (SNS/PN3)-null mice. J. Neurosci. 22:8352-8356.
    Lanteri-Minet, M., Bon, K., de Pommery, J., Michiels, J.F., and Menetrey, D. 1995. Cyclophosphamide cystitis as a model of visceral pain in rats: Model elaboration and spinal structures involved as revealed by the expression of c-Fos and Krox-24 proteins. Exp. Brain Res. 105:220-232.
    Olivar, T. and Laird, J.M. 1999. Cyclophosphamide cystitis in mice: Behavioral characterization and correlation with bladder inflammation. Eur. J. Pain 3:141-149.
    Wantuch, C., Piesla., M.P., and Leventhal, L. 2007. Pharmacological validation of a model of cystitis pain in the mouse. Neurosci. Lett. 421:250-252.
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