The biological processes that mediate and modulate the perception of pain in the infant animal are not well studied and thus nociception during early development is poorly understood. In the adult animal, injection of formalin into the hind paw produces distinct phases of behavioral and autonomic responses: an early nociceptive response followed by a period of quiescence and a later second phase that matches or exceeds the initial response. The delayed reaction of the second phase has been suggested to be a model of inflammation-induced changes in neuronal sensitivity. Studies in the infant rat have demonstrated that the first phase is present in the fetus and neonate but the onset of the second phase is later maturing. We report here that the first phase occurs in 7- to 35-day-old pups in the formalin test when measured behaviorally and in 14- to 35-day-old pups when assessed by increased heart rate. However, the behavioral response in second phase is greatly attenuated or absent in 7- or 14-day-old pups? a finding consistent with that of others, appearing first at 21 days of age. The biphasic tachycardic response was not noted until even later, at 35 days of age. These data confirm that the neural mechanisms that mediate the secondary behavioral phase in the formalin test are late maturing, that the biphasic cardiovascular response does not occur until substantially later, after weaning, and that the behavioral and cardiovascular responses are dissociated developmentally. (C) 1998 Elsevier Science Inc.