Rodent Models of Colorectal Distension

Siobhain M. O'Mahony1, Monica Tramullas1, Patrick Fitzgerald1, John F. Cryan1

1 Alimentary Pharmabiotic Centre, Biosciences Institute, University College Cork, Cork, Ireland
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 9.40
DOI:  10.1002/0471142301.ns0940s61
Online Posting Date:  October, 2012
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Colorectal distension (CRD) is a widely accepted, reproducible method for assessing visceral sensitivity in both clinical and pre‐clinical studies. Distension of the colon mirrors the human scenario of visceral pain with regard to intensity and referral of pain in patients. There are several readouts that can be applied to the CRD protocol depending on the species being evaluated, two of which are described in this unit. CRD can be used to measure the impact of novel compounds, strain, or genetic differences as well as the effect of physical and psychological stressors on the sensitivity of the colon. Investigation of the impact of a noxious visceral stimulus (CRD) on other systems within the body can also be carried out. Given that visceral pain is a major clinical problem and one of the most common reasons patients seek out medical advice, the ability to assess this type of pain is essential to the discovery of successful treatments. This unit outlines two protocols that describe CRD of rats and mice. Curr. Protoc. Neurosci. 61:9.40.1‐9.40.13. © 2012 by John Wiley & Sons, Inc.

Keywords: visceral hypersensitivity; rodent models of pain; colorectal distension; abdominal withdrawal reflex; visceromotor response

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Basic Protocol 1: Abdominal Withdrawal Reflex During Colorectal Distension in Rats
  • Support Protocol 1: Construction of Balloons for Use in Rats
  • Basic Protocol 2: Assessment of Visceral Pain Responses to Colorectal Distension in Mice
  • Support Protocol 2: Construction of Balloon Used in Mice
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: Abdominal Withdrawal Reflex During Colorectal Distension in Rats

  Materials
  • Male Sprague‐Dawley rats (at least 250 g and ∼10 to 11 weeks old)
  • Anesthetic (Isoflurane; IsoFlo, Abbott)
  • Water‐soluble and non‐irritating lubricant jelly (Johnson & Johnson)
  • Home cages
  • Barostat (Distender Series II, G&J Electronics; http://www.barostat.com/rat.html)
  • Personal computer (P.C.) and Protocol Plus software (G&J Electronics)
  • Polyethylene tubing (PE60; 0.76‐mm i.d., 1.22‐mm o.d.; Becton Dickinson)
  • Fasting cages
  • Vaporizer unit (Bartons)
  • Balloons (see protocol 2)
  • Surgical hypoallergenic tape (∼25‐mm, Micropore)
  • Zinc oxide stretch hypoallergenic tape (∼25‐mm, Fleming)
  • Recovery cages
  • Plexiglas observation chamber (40 × 28 × 54–cm)

Support Protocol 1: Construction of Balloons for Use in Rats

  Materials
  • Vygon intravenous catheter with luer‐lock at one end (16‐G, 30‐cm; Vygon)
  • Ruler
  • Marker
  • No. 22 scalpel blade (Fine Science Tools)
  • Extra‐safe latex condoms (Durex)
  • 4‐0 silk suture (Ethicon, cat. no. W329H)
  • 10‐ml syringe (Becton Dickenson)

Basic Protocol 2: Assessment of Visceral Pain Responses to Colorectal Distension in Mice

  Materials
  • Adult mice, either sex, at least 6 weeks of age (21 to 23 g)
  • Anesthetic (isoflurane; IsoFlo, Abbott)
  • Water‐soluble and non‐irritating lubricant jelly (Johnson & Johnson)
  • Personal computer (P.C.) and Protocol Plus software (G&J Electronics)
  • Barostat (Distender Series II, G&J Electronics)
  • Transducer amplifier (LabTrax 4, World Precision Instruments)
  • Vaporizer unit (Bartons)
  • Cotton‐tipped applicators
  • Balloon (see protocol 2 or 2)
  • Surgical hypoallergenic tape (∼25 mm, Micropore)
  • Zinc oxide stretch hypoallergenic tape (∼25 mm, Fleming)
  • Data Trax 2 software (World Precision Instruments)
  • Statistic software (SPSS or GraphPad Prism)

Support Protocol 2: Construction of Balloon Used in Mice

  Materials
  • Polyethylene tubing (PE60; I.D = 0.76‐mm i.d., 1.22–mm o.d., Becton Dickinson)
  • 27‐G sterile hypodermic needle (BD, cat. no. 300635)
  • Custom‐made balloons (2‐cm length × 1‐cm inflated diameter, GMC Medical)
  • 4‐0 silk suture (Ethicon, cat. no. W329H)
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Al‐Chaer, E.D., Feng, Y., and Willis, W.D. 1998. A role for the dorsal column in nociceptive visceral input into the thalamus of primates. J. Neurophysiol. 79:3143‐3150.
   Al‐Chaer, E.D., Kawasaki, M., and Pasricha, P.J. 2000. A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development. Gastroenterology 119:1276‐1285.
   Arvidsson, S., Larsson, M., Larsson, H., Lindstrom, E., and Martinez, V. 2006. Assessment of visceral pain‐related pseudo‐affective responses to colorectal distension in mice by intracolonic manometric recordings. J. Pain 7:108‐118.
   Cevese, A., Mary, D.A., Poltronieri, R., Schena, F., and Vacca, G. 1992. Haemodynamic effects of distension of the descending colon in anaesthetized dogs. J. Physiol. 447:409‐423.
   Christianson, J.A. and Gebhart, G.F. 2007. Assessment of colon sensitivity by luminal distension in mice. Nat. Protoc. 2:2624‐2631.
   Coutinho, S.V., Plotsky, P.M., Sablad, M., Miller, J.C., Zhou, H., Bayati, A.I., McRoberts, J.A., and Mayer, E.A. 2002. Neonatal maternal separation alters stress‐induced responses to viscerosomatic nociceptive stimuli in rat. Am. J. Physiol. Gastrointest. Liver Physiol. 282:G307‐G316.
   Crawford, M.E., Jensen, F.M., Toftdahl, D.B., and Madsen, J.B. 1993. Direct spinal effect of intrathecal and extradural midazolam on visceral noxious stimulation in rabbits. Br. J. Anaesth. 70:642‐646.
   Gibney, S.M., Gosselin, R.D., Dinan, T.G., and Cryan, J.F. 2010. Colorectal distension‐induced prefrontal cortex activation in the Wistar‐Kyoto rat: Implications for irritable bowel syndrome. Neuroscience 165:675‐683.
   Gosselin, R.D., O'Connor, R.M., Tramullas, M., Julio‐Pieper, M., Dinan, T.G., and Cryan, J.F. 2010. Riluzole normalizes early‐life stress‐induced visceral hypersensitivity in rats: role of spinal glutamate reuptake mechanisms. Gastroenterology 138:2418‐2425.
   Guthrie, E., Barlow, J., Fernandes, L., Ratcliffe, J., Read, N., Thompson, D.G., Tomenson, B., and Creed, F. 2004. Changes in tolerance to rectal distension correlate with changes in psychological state in patients with severe irritable bowel syndrome. Psychosom. Med. 66:578‐582.
   Kamp, E.H., Jones, R.C., 3rd, Tillman, S.R., and Gebhart, G.F. 2003. Quantitative assessment and characterization of visceral nociception and hyperalgesia in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 284:G434‐G444.
   Larauche, M., Gourcerol, G., Million, M., Adelson, D.W., and Tache, Y. 2010. Repeated psychological stress‐induced alterations of visceral sensitivity and colonic motor functions in mice: Influence of surgery and postoperative single housing on visceromotor responses. Stress 13:343‐354.
   Larsson, M., Arvidsson, S., Ekman, C., and Bayati, A. 2003. A model for chronic quantitative studies of colorectal sensitivity using balloon distension in conscious mice Effects of opioid receptor agonists. Neurogastroenterol. Motil. 15:371‐381.
   Martinez, V. and Melgar, S. 2008. Lack of colonic‐inflammation‐induced acute visceral hypersensitivity to colorectal distension in Na(v)1.9 knockout mice. Eur. J. Pain 12:934‐944.
   Ness, T.J. and Gebhart, G.F. 1988. Colorectal distension as a noxious visceral stimulus: Physiologic and pharmacologic characterization of pseudaffective reflexes in the rat. Brain Res. 450:153‐169.
   Nijsen, M.J., Ongenae, N.G., Coulie, B., and Meulemans, A.L. 2003. Telemetric animal model to evaluate visceral pain in the freely moving rat. Pain 105:115‐123.
   O'Mahony, S.M., Marchesi, J.R., Scully, P., Codling, C., Ceolho, A.M., Quigley, E.M., Cryan, J.F., and Dinan, T.G. 2009. Early life stress alters behavior, immunity, and microbiota in rats: Implications for irritable bowel syndrome and psychiatric illnesses. Biol. Psychiatry 65:263‐267.
   Tramullas, M., Dinan, T.G., and Cryan, J.F. 2011. Chronic psychosocial stress induces visceral hyperalgesia in mice. Stress 15:281‐292.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library