The formalin test: a dose-response analysis at three developmental stages

We investigated the behavioral response of rat pups to intraplantar injection of varying formalin concentrations using a time-sampling method. At 3 days of age, the response was monophasic and persisted for the whole hour, even at low formalin concentrations. Flexion, shaking and licking the injected limb and hind-limb kicking correlated strongly with log formalin concentration (r = 0.82); behavioral state was altered only at the highest concentration. The response on day 15 was also monophasic, but it waned in 30 min, even at the highest formalin concentration tested. Flexion, shaking and licking of the injected limb were strong pain measures (r = 0.83). The response at 25 days was biphasic, and the adult measures, paw lifting and licking, produced a good formalin concentration-effect relationship (r = 0.80). The log concentration-effect relationships for formalin at the three developmental stages and for adult rats were parallel, but between 3 days and 15 days of age, the relationship shifted to the right by 2.5-fold, and by a further 4-fold between 15 and 25 days, when the sensitivity to formalin-induced pain was similar to that in adults. The data describe efficient, quantitative measures of formalin-induced pain for developing rats, show that the pain response is log-linearly related to formalin concentration throughout development, and demonstrate that the sensitivity to formalin-induced pain is about 10-fold higher in neonatal rats than in weanlings. the data imply that there are major qualitative changes in pain processing as the nervous system develops.