The expression of the immediate early gene c-fos has been used extensively as a marker for neural activation in response to acute and chronic stressful stimuli in brain and spinal cord. The present study examined the expression of Fos protein in the brainstem nuclei of male and female gonadectomized mice in response to cold stress. Free-floating sections were processed immunohistochemically for Fos protein using standard avidin-biotin complex methods. The number of Fos-positive neurons in each nucleus was determined. Although the experiment was designed to look for gender differences, results were equivalent between females and males. After mice were exposed to a cold ambient temperature (4 degrees C) for 2 h, elevated numbers of Fos-positive neurons were counted in the medullary gigantocellular reticular nucleus, medullary raphe nucleus, nucleus of the solitary tract and in locus coeruleus. However, no elevated expression was found in nucleus ambiguus, nor in neurons of the A1 group, nor the C2 or C3 group. Similar to rats, these results with mice reveal a widespread arousal system in the lower brainstem activated by cold stress in both genders. These findings in gonadectomized mice have set the stage for investigations following hormonal and genetic manipulations.