Models of Nociception: Hot‐Plate, Tail‐Flick, and Formalin Tests in Rodents
Experimental models of pain include tests of response thesholds to high intensity stimuli (acute pain tests) and changes in spontaneous or evoked behavioral responses in animals with peripheral injury or inflammation (persistent pain models). Acute thermal pain is modeled by the hot‐plate and tail‐flick test, while persistent pain can be modeled by the formalin test. This unit presents protocols for all three of these tests, including preparation of animals (rats or mice), administration of a compound being tested for its analgesic properties and data collection. Curr. Protoc. Neurosci. 41:8.9.1‐8.9.16. © 2007 by John Wiley & Sons, Inc.
Keywords: pain; nociception; tail flick; formalin; hot plate; rat; mouse; animal model; analgesia
Table of Contents
- Basic Protocol 1: Measurement of Acute Pain Using the Hot‐Plate Test
- Basic Protocol 2: Measurement of Acute Pain Using the Tail‐Flick Test
- Basic Protocol 3: Measurement of Persistent Pain Using the Formalin Test
- Reagents and Solutions
- Literature Cited
Basic Protocol 1: Measurement of Acute Pain Using the Hot‐Plate Test
Basic Protocol 2: Measurement of Acute Pain Using the Tail‐Flick Test
Basic Protocol 3: Measurement of Persistent Pain Using the Formalin Test
Figure 8.9.1 Apparatus used for the tail‐flick test.
Figure 8.9.2 Apparatus used for formalin test.
Figure 8.9.3 Change in pain score through phases 1 and 2 in an animal injected with formalin.
Figure 8.9.4 Effects of morphine on paw lick and jump latency in the hot‐plate assay in mice. Separate experiments were conducted for each endpoint.
Figure 8.9.6 Effect of morphine in the tail flick test in mice. The mice were injected subcutaneously (s.c.) 30 min before the tail‐flick test. n=5 to 8 animals per dose.
|Bannon, A.W., Gunther, K.L., and Decker, M.W. 1995. Is epibatidine really analgesic? Dissociation of the locomotor activity, temperature, and analgesic effects of (±)‐epibatidine. Pharmacol.Biochem. Behav. 51:693‐698.|
|Berge, O.‐G., Garcia‐Cabrera, I., and Hole, K. 1988. Response latencies in the tail‐flick test depend on tail skin temperature. Neurosci. Lett. 86:284‐288.|
|D'Amour, F.E. and Smith, D.L. 1941. A method for determining loss of pain sensation. J. Pharmacol. Exp. Ther. 41:419‐424.|
|Dickenson, A.H. and Sullivan, A.F. 1987. Peripheral origins and central modulation of subcutaneous formalin‐induced activity of rat dorsal horn neurons. Neurosci. Lett. 83:207‐211.|
|Dubuisson, D. and Dennis, S.G. 1977. The formalin test: A quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats. Pain 4:161‐174.|
|Hargreaves, K., Dubner, R., Brown, F., Flores, C., Jorrs, J. 1988. A new sensitive method for measuring thermal nociception in cutaneous hyperalgesia Pain 32:77‐88.|
|Hunskaar, S., Fasmer, O.B., and Hole, K. 1985. Formalin test in mice, a useful technique for evaluating mild analgesics. J.Neurosci. Methods 14:69‐76.|
|Janssen, P.A.J., Niemegeers, C.J.E., and Dony, J.G.H. 1963. The inhibitory effect of fentanyl and other morphine‐like analgesics on the warm water induced tail withdrawal reflex. Arzneim. Forsch. 13:502‐507.|
|Jett, M.F. and Michelson, S. 1996. The formalin test in rat: Validation of an automated system. Pain 64:19‐25.|
|Jourdan, D., Ardid, D., Bardin, L., Bardin, M., Neuzeret, D., Lanphouthacoul, L., and Eschalier, A. 1997. A new automated method of pain scoring in the formalin test in rats. Pain 71:265‐270.|
|Jourdan, D., Alloui, A., and Eschalier, A. 1999. Pharmacological validation of an automated method of pain scoring in the formalin test in rats. J. Pharmacol. Toxicol. Methods 42:163‐170.|
|Malkmus, S., Lu, X., Bartfai, T., Yaksh, T.L., and Hua, X.Y. 2005. Increased hyperalgesia after tissue injury and faster recovery of allodynia after nerve injury in the GalR1 knockout mice. Neuropeptides 39:217‐221.|
|Mogil, J.S., Kest, B., Sadowski, B., and Belknap, J.K. 1996. Differential genetic mediation of sensitivity to morphine in genetic models of opiate antinociception: Influence of nociceptive assay. J. Pharmacol. Exp. Ther. 276:532‐544.|
|Murray, C.W., Porreca, F., and Cowan, A. 1988. Methodological refinements to the mouse paw formalin test. An animal model of tonic pain. J. Pharmacol. Methods 20:175‐186.|
|Sufka, K.J., Watson, G.S., Nothdurft, R.E., and Mogil, J.S. 1998. Scoring the mouse formalin test: Validation study. Eur. J. Pain 2:351‐358.|
|Tjøsen, A., Lund, A., Berge, O.‐G., and Hole, K. 1989. An improved method for tail‐flick testing with adjustment for tail‐skin‐temperature. J. Neurosci. Methods 26:259‐265.|
|Wheeler‐Aceto, H. and Cowan, A. 1991. Standardization of the rat paw formalin test for the evaluation of analgesics. Psychopharmacol. 104:35‐44.|
|Yaksh, T.L., Ozaki, G., McCumber, D., Rathbun, M., Svensson, C., Malkmus, S., and Yaksh, M.C. 2001. An automated flinch detecting system for use in the formalin nociceptive bioassay. J. Appl. Physiol. 90:2386‐23402.|