Models of Muscle Pain: Carrageenan Model and Acidic Saline Model

Rajan Radhakrishnan1, Marie K. Hoeger Bement1, David Skyba1, Kathleen A. Sluka1, Lois J. Kehl2

1 University of Iowa, Iowa City, Iowa, 2 University of Minnesota, Minneapolis, Minnesota
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.35
DOI:  10.1002/0471141755.ph0535s25
Online Posting Date:  September, 2004
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Abstract

Carrageenan or acidic saline injected unilaterally into the gastrocnemius muscle or triceps muscle produces a robust and long‐lasting hyperalgesia in rats and mice, which is reversible with systemic administration of opioid or anti‐inflammatory drugs. This unit describes detailed protocols for inducing and measuring hyperalgesia, and provides information on validation of these models. These models are useful for assessing new compounds for their analgesic activity in muscular pain.

Keywords: Muscle; Hyperalgesia; Carrageenan; Acidic saline; Rats; Mice

     
 
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Table of Contents

  • Basic Protocol 1: Carrageenan/Grip‐Force Model of Muscle Pain in Rats
  • Alternate Protocol 1: Carrageenan/Grip‐Force Model of Muscle Pain in Mice
  • Basic Protocol 2: Carrageenan Model of Muscle Pain in Rats
  • Alternate Protocol 2: Carrageenan Model of Muscle Pain in Mice
  • Basic Protocol 3: Acidic Saline Model of Muscle Pain in Rats
  • Alternate Protocol 3: Acidic Saline Model of Muscle Pain in Mice
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Carrageenan/Grip‐Force Model of Muscle Pain in Rats

  Materials
  • Sprague‐Dawley male rats (100 to 150 g, Harlan; see for considerations regarding age, strain and gender)
  • Cylinder of compressed medical oxygen
  • Halothane USP
  • Povidone‐iodine solution
  • 5.3% (w/v) lambda carrageenan, type IV (Sigma) in normal sterile 0.9% saline adjusted to pH 7.0 (see recipe), prepare fresh just before use
  • Reference analgesic compound, i.e., positive control, (e.g., levorphanol tartrate salt, indomethacin, dexamethasone 21‐phosphate disodium salt; see recipe)
  • Animal scale (accurate to 1 g)
  • Grip‐force analyzer with DFIS series force transducer and attached wire mesh grid (12 × 7–cm2 with wire grid 1.2 cm apart) to measure tensile force with minimum and maximum detection limits of 20 g and 1 kg, respectively (Chatillon)
  • Anesthesia induction equipment, which includes a Plexiglas anesthetic chamber (24 × 10 × 10–cm) connected to a halothane vaporizer (Datex‐Ohmeda) with Y‐connectors and polythene tubing to allow nose cone adapter connections
  • Sterile absorbent pads (60 × 40–cm)
  • Nose cone adapter
  • 1‐ml sterile disposable tuberculin syringes
  • 25‐G sterile disposable hypodermic needles
  • Electronic timer (with count‐up and count‐down capability)

Alternate Protocol 1: Carrageenan/Grip‐Force Model of Muscle Pain in Mice

  • C3H/HeJ mice (23 to 28 g; National Cancer Institute)
  • 4% (w/v) lambda carrageenan type IV (Sigma) in normal sterile 0.9% saline adjusted to pH 7.0 (see protocol 2)
  • Reference analgesic compound (positive control), e.g., morphine sulfate (Sigma)
  • Wire mesh grid attachment for force transducer with smaller size grid (12 × 7–cm2 with wire grid 0.5 cm apart) for mice
  • 0.5‐ml sterile, disposable insulin syringes equipped with 29‐G needles

Basic Protocol 2: Carrageenan Model of Muscle Pain in Rats

  Materials
  • Male Sprague‐Dawley rats (250 to 300 g; Harlan)
  • Halothane USP
  • Cylinder of compressed medical oxygen
  • Povidone‐iodine solution
  • 3% (w/v) lambda carrageenan, type IV (Sigma) in normal sterile saline adjusted to pH 7.0 (see recipe)
  • Sterile normal saline adjusted to pH 7.0
  • Reference analgesic compound (positive control), e.g., morphine sulfate (morphine solution for rats, see recipe)
  • Animal weighing scale (accurate to 1 g)
  • Plexiglas cubicles (16 × 8 × 8–cm) with open bottom and ventilation holes in the walls
  • Platform with glass top (75 × 37 × 17–cm; Fig. B)
  • Radiant heat source (halogen lamp, 35 W) powered with a variable (0 to 140 V) but consistent voltage power supply and with a built‐in timer
  • Platform with wire mesh top (75 × 37 × 30–cm; Fig. C)
  • Set of von Frey filaments ranging from 8 mN to 350 mN (8, 12, 16, 32, 44, 56, 75, 104, 162, and 350 mN; North Coast Medical)
  • Anesthesia induction equipment, which includes a Plexiglas anesthetic chamber (24 × 10 × 10–cm) connected to a halothane vaporizer (Datex‐Ohmeda)
  • Sterile absorbent pads (60 × 40–cm)
  • Nose cone adapter
  • 1‐ml sterile, disposable syringes
  • 25‐G sterile, disposable hypodermic needles

Alternate Protocol 2: Carrageenan Model of Muscle Pain in Mice

  • C57BL/6J male mice (25 to 30 g; Jackson Laboratories)
  • Morphine solution for mice (see recipe)
  • Plexiglas cubicles (9 × 3 × 5–cm) with open bottom and ventilation holes in the walls
  • Radiant heat source (halogen lamp, 35 W) powered with a variable (0 to 140 V) but consistent voltage power supply, with a small (e.g., ∼5 × 5–mm) aperture for the heat source and with a built‐in timer
  • von Frey filament with 0.4 mN (0.04 g) bending force (Touch Test, Stoelting)
  • 0.5‐ml sterile, disposable syringes
  • 28‐G sterile, disposable hypodermic needles

Basic Protocol 3: Acidic Saline Model of Muscle Pain in Rats

  Materials
  • Male Sprague‐Dawley rats (250 to 300 g; Harlan)
  • 0.01 N sterile NaOH
  • 0.01 N sterile HCl
  • Preservative‐free normal saline
  • Halothane USP
  • Cylinder of compressed medical oxygen
  • Reference analgesic compound (positive control) e.g., morphine sulfate (morphine solution for rats, see recipe)
  • Plexiglas cubicles (16 × 8 × 8–cm) on an elevated wire mesh
  • 0.5‐ml sterile, disposable syringes
  • 28‐G sterile, disposable hypodermic needles
  • Set of von Frey filaments (8, 12, 16, 32, 44, 56, 75, 104, 162, 350 mN; North Coast Medical)
  • Anesthesia induction equipment including Plexiglas anesthetic chamber (24 × 10 × 10–cm) connected to a halothane vaporizer (Datex‐Ohmeda)

Alternate Protocol 3: Acidic Saline Model of Muscle Pain in Mice

  • C57BL/6J male mice (25 to 30 g; Jackson Laboratories)
  • Morphine solution for mice (see recipe)
  • Plexiglas cubicles (9 × 3 × 5–cm)
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Figures

Videos

Literature Cited

Literature Cited
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