Assessment of Gastrointestinal Propulsive Activity Using Three Different Models of Peristalsis In Vivo in the Mouse

Enzo Poli1, Cristina Pozzoli1

1 University of Parma Medical School, Parma, Italy
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 21.9
DOI:  10.1002/0471140856.tx2109s46
Online Posting Date:  November, 2010
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Abstract

The protocols described in this unit are designed to assess the acute effects of drugs on the propulsive activity of the gastrointestinal muscles in the conscious mouse. These protocols are currently applied to investigate the pharmacological activity of novel compounds undergoing preclinical development and to obtain predictive data needed to advance drugs into clinical trials. Moreover, these methods could be useful in evaluating the functional toxicity by environmental or alimentary pollutants, like xenobiotics and naturally occurring toxins endowed with noxious activity in the control of physiologic peristalsis. The three models detailed—the measurement of gastric emptying, ileal transit, and colonic propulsion—are substantially non‐invasive and do not require analgesic pretreatments or the induction of general anesthesia. In contrast to an in vitro approach, these in vivo studies provide a unified understanding of drug effects on gut functionality, in particular when the central nervous system, the extrinsic nerves, or the (neuro)endocrine system is targeted by the test drugs.Curr. Protoc. Toxicol. 46:21.9.1‐21.9.20. © 2010 by John Wiley & Sons, Inc.

Keywords: gastrointestinal tract; peristaltic activity; gastric emptying; intestinal transit; colonic transit; gut propulsion

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Measurement of Gastric Emptying of Solid Nutrients
  • Basic Protocol 2: Measurement of Gastrointestinal Transit with Charcoal
  • Basic Protocol 3: Measurement of Colonic Propulsion
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Measurement of Gastric Emptying of Solid Nutrients

  Materials
  • Male Swiss mice, ∼70 days old, ∼30 to 35 g of body weight (before fasting)
  • Standard mice pellet diet (e.g., Harlan Laboratories Teklad Global Diet)
  • Test compound(s) (see recipe)
  • Atropine sulfate (see recipe)
  • Clonidine hydrochloride (positive control alternative to atropine; see recipe)
  • (2‐hydroxypropyl)‐β‐cyclodextrin (HBC), 20% (w/v) solution (optional, to be used when lipophilic drugs do not dissolve freely in physiological saline; see recipe)
  • Physiological saline: 0.9 g NaCl in 1 liter of distilled water
  • Diethyl ether
  • Polypropylene cages, with an approximate floor area 1000 cm2 and a wire mesh grid to fit the bottom, to house animals before experiments
  • Latex gloves
  • Animal balance (accuracy ±0.1 g; e.g., Mettler)
  • Permanent waterproof markers, one color for each test group
  • 1‐ml plastic syringes (e.g., PIC Indolor), with 0.01‐ml scale (tuberculin syringes) equipped with 26‐G needles
  • Chronometer (for recording treatment times)
  • Polypropylene cages for single animals, with an approximate floor area 150 to 200 cm2 and a wire mesh grid to fit the bottom
  • Metabolic cages (as an optimum), adapted to measure food and water intake in mice, suitable also for measuring the metabolism of the animal
  • Calculator
  • Glass bell (25‐cm diameter, total volume 3 to 5 liter)
  • Hydrophilic cotton (standard product used for medications)
  • Surgical equipment including:
    • Two small hemostats (curved forceps 10‐cm long; Rudolf Surgical Instruments)
    • Curved hemostat (14‐cm long; Rudolf Surgical Instruments)
    • Straight scissors (10‐cm long; Rudolf Surgical Instruments)
    • Curved scissors (14.5‐cm long; Rudolf Surgical Instruments)
  • 50‐ml plastic beakers
  • Paper towels

Basic Protocol 2: Measurement of Gastrointestinal Transit with Charcoal

  Materials
  • Male Swiss mice, ∼70 days old, 30 to 35 g of body weight (before fasting)
  • Standard mice pellet diet (e.g., Harlan Laboratories Teklad Global Diet)
  • Test compound(s) (see recipe)
  • Reference compounds (to be used as controls in step 6):
    • Atropine sulfate (Sigma; see recipe)
    • Clonidine hydrochloride (Sigma; see recipe)
    • Bethanechol hydrochloride (Sigma; see recipe)
  • Charcoal suspension (see recipe)
  • (2‐hydroxypropyl)‐β‐cyclodextrin (HBC), 20% (w/v) solution (optional; see recipe), to be used when lipophilic drugs do not dissolve freely in physiological saline (HBC should be employed as vehicle in control experiments)
  • Physiological saline: dissolve 0.9 g/liter NaCl (analytical grade) in distilled water
  • Diethyl ether
  • Polypropylene cages, with an approximate floor area 1000 cm2 and a wire mesh grid to fit the bottom of the cage
  • Latex gloves
  • Animal scale accuracy ±0.1 g; e.g., Mettler
  • Permanent waterproof markers, one color for each group
  • 1‐ml plastic syringes (e.g., PIC Indolor), with 0.01‐ml scale (tuberculin syringes) equipped with 26‐G needles
  • Chronometer
  • Calculator
  • Plastic cannula 1‐mm outer diameter, 6‐ to 7‐cm long
  • 2.5‐ml plastic syringes, graduation 0.1 ml
  • 50‐ml plastic beakers
  • Glass bell (25‐cm diameter, total volume 3 to 5 liter)
  • Hydrophilic cotton
  • Hemostat (curved forceps 14‐cm long; Rudolf Surgical Instruments)
  • Two curved hemostats (10‐cm long; Rudolf Surgical Instruments)

Basic Protocol 3: Measurement of Colonic Propulsion

  Materials
  • Male Swiss mice, ∼70 days old, 25 to 30 g of body weight (before fasting)
  • Standard mice pellet diet (e.g., Harlan Laboratories Teklad Global Diet)
  • Test compound(s) (see recipe)
  • Loperamide hydrochloride (see recipe)
  • (2‐hydroxypropyl)‐β‐cyclodextrin (HBC), 20% (w/v) solution (optional; see recipe), to be used when lipophylic drugs do not dissolve freely in physiological saline (HBC should be employed as vehicle in control experiments)
  • Physiological saline: 0.9 g NaCl in 1 liter of distilled water
  • Diethyl ether
  • Polypropylene cages, with an approximate floor area 1000 cm2 and a wire mesh grid to fit the bottom, to house animals before experiments
  • Latex gloves
  • Animal scale (accuracy ±0.1 g, e.g., Mettler)
  • Permanent waterproof markers, one color for each test group
  • 1‐ml plastic syringes, (e.g., PIC Indolor, Artsana), 0.01‐ml scale (tuberculin syringes) equipped with 26‐G needles
  • 50‐ml plastic beakers
  • Glass bell (25‐cm diameter, total volume 3 to 5 liter)
  • Hydrophilic cotton (standard product used for medications)
  • Chronometer for recording time
  • Glass bead, 2‐mm wide, 3‐mm long, fire‐polished to avoid mucosal damage, resembling, in form and dimension, the normal mice stool
  • Glass rod, 1‐mm caliber, with fire‐polished tip, to avoid colonic damage when used to insert beads into the rectum, marked at a distance of 2‐cm from the tip with a permanent marker
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Figures

Videos

Literature Cited

Literature Cited
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   Megumi, T., Song, C.W., Park, Y.G., Tanaka, Y., Uchida, I., and Okumoto, M. 2004. Mapping of putative ether‐anesthesia resistance gene using C57BL/6J and MSM/Ms mouse strains. J. Anesthesiol. 18:267‐276.
   Pozzoli, C., Todorov, S., Schunack, W., Timmerman, H., Coruzzi, G., and Poli, E. 2002. Role of histamine H3 receptors in control of mouse intestinal motility in vivo and in vitro. Comparison with α2‐adrenoceptors. Dig. Dis. Sci. 47:1065‐1072.
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