Expression of the c-fos gene in CNS induced by formalin injection into the face as pain stimulus was examined in cats. Fos-positive neurons were demonstrated in the anterior cingulate, anterior insula and other areas of the cerebral cortex bilaterally, midline thalamic nuclei, hypothalamus, and brainstem in both the formalin-injected and control group (anesthesia only). Most of these labeled regions appeared to correspond to stress- and anesthesia-related sites. The difference from the control was the finding of Fos-positive cells in the SI, possible SII and trigeminal subnucleus caudalis in the experimental group. Almost significant increase in Fos-positive cells was also observed in areas 24, 23 and the anterior agranular insular cortex in experimental cats. Our findings appear to be compatible with recently reported PET findings in man, except laterality. For full appreciation of the complex pain experience, these diverse areas of the brain appear to be activated.