To investigate the role of the corpus callosum in the expression of functional brain asymmetries, we compared left and right uptake of [14C]2-deoxyglucose in 43 brain regions measured in 10 C57B1/6 mice with a normal corpus callosum and in 12 congenitally acallosal mice, after 45 min of free activity in a novel, large open-field arena. The metabolic patterns across the brain appeared to be similar in the two groups of mice, as well as the average direction of asymmetry in tracer incorporation, which was higher at right in most of the brain regions for both acallosals and controls. However, the direction of the metabolic asymmetries of any given region was not consistent across individual animals. The largest asymmetries were found in the central auditory nuclei in both groups of mice, with extreme values in some acallosals. Significantly larger asymmetries were found in acallosal mice for the brain and the cortex as a whole, as well as for the lateral geniculate and pretectal nuclei, the olfactory tubercles, and retrosplenial, infrarhinal and perirhinal cortices. The metabolic asymmetries of the thalamic sensory nuclei were correlated with the asymmetries of the corresponding sensory cortical fields in the acallosal, but not in control mice. On the other hand, asymmetries of the cortical regions were largely intercorrelated in control mice, resulting in a general activation of one hemisphere over the other, while in acallosals they were more independent, resulting in a "patchy" pattern of cortical asymmetries. These results suggest that callosal agenesis, combined with the occurrence of ipsilateral Probst bundles, leads to a loss of co-ordination in the activation of different sensory and motor areas. The impaired co-ordination might then be distributed through cortico-subcortical loops, resulting in larger asymmetries throughout the brain. Thus, a normal corpus callosum appears to balance and synchronize metabolic brain activity, perhaps by smoothing the effects of asymmetrically activated ascending systems.