Collection of Bile and Urine Samples for Determining the Urinary and Hepatobiliary Disposition of Xenobiotics in Mice

José E. Manautou1, Chuan Chen1

1 University of Connecticut, Storrs, Connecticut
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 5.7
DOI:  10.1002/0471140856.tx0507s22
Online Posting Date:  January, 2005
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Abstract

One of the most simple and informative method commonly used by researchers in the field of xenobiotics metabolism and dispostion to investigate the fate of chemicals in living organisms involves identification and/or quantitation of metabolites in excreta such as urine and bile. This method can be performed on dogs and larger rodents such as rats and hamsters rather easily. However, many investigators find it difficult to collect bile samples from mice. This unit describes, in detail, how to collect urine and bile samples from a mouse for further metabolite analysis.

     
 
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Table of Contents

  • Basic Protocol 1: Collection of Bile and Urine Samples from a Bile Duct–Cannulated Mouse
  • Alternate Protocol 1: Collection of Bile Samples from Gall Bladder–Cannulated Mouse
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Collection of Bile and Urine Samples from a Bile Duct–Cannulated Mouse

  Materials
  • 0.9% saline (NaCl), sterile
  • Mice (3 to 4 months old or body weight >25 g)
  • Paraffin oil
  • Ketamine and xylazine mixture or other appropriate anesthetics
  • Test compound dissolved in appropriate sterile vehicle
  • Electric clippers
  • Scalpel blades
  • Cotton‐tipped applicators
  • Surgical microscope or appropriate magnifier
  • 3‐0 silk sutures
  • Microdissecting forceps
  • Hemostatic forceps
  • 1.2‐French polyurethane tubing (Access Technologies)
  • Gauze pads
  • Temperature‐control system composed of the following:
    • Rectal thermistor probe for rodents
    • Heating lamp
    • Homeothermic control unit
  • 1.5‐ml microcentrifuge tubes
  • 1‐ml polypropylene disposable syringe
  • 30‐ and 25‐G needles
  • Analytical balance
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Figures

  •   FigureFigure 5.7.1 Diagram of the bile duct system in mice. Figure reproduced from Iwaki et al., () with permission of Braintree Scientific, Inc.
  •   FigureFigure 5.7.2 Effect of the non‐metabolizable organic anion dibromosulphothphalein (DBSP) on bile flow rate (A); biliary concentration of acetaminophen‐glutathione conjugate (APAP‐GSH) (B); and the cumulative excretion of APAP and its metabolites (C). Following overnigt (18 hr) fasting, adult male CD‐1 mice were anesthetized and the common bile duct was cannulated. Bile was collected for 15 min before mice received 120 µmol DBSP/kg immediately prior to APAP (1 mmol/kg, i.v.). Controls received saline vehicle before APAP. After APAP dosing, bile was collected for 150 min at 30‐min intervals. Results are expressed as means ±SEM for 4 to 5 animals per group. *Significantly different from controls ( p < 0.05).

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Literature Cited

Literature Cited
   Iwaki, T., Yamashita, H., and Hayakawa, T. 2001. A Color Atlas of Sectional Anatomy of the Mouse. Braintree Scientific, Inc. Braintree, Mass.
   Kanz, M.F., Whitehead, R.F., Freguson, A.E., Kaphalia, L., and Moslen, M.T. 1992. Biliary function studies during multiple time periods in freely moving rats. A useful system and set of marker solutes. J. Pharmacol. Toxicol. Methods 27:7‐15.
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