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The midsagittal area of the corpus callosum was measured in its entirety and in seven subdivisions in a sample of 50 brains consecutively obtained from autopsies of individuals who had neuropsychological testing before death. A 12-item test of hand preference was used as an index of the pattern of interhemispheric functional asymmetry. Callosal size was analysed for two factors: hand preference, classified as consistent-right-hand preference (CRH) versus non consistent-right-hand preference, and sex. The group of nCRH (n = 18) was found to have a larger overall callosal area, with the greatest difference occurring in the posterior body segments, especially the isthmus. The isthmus probably includes interhemispheric fibres from posterior parietal and superior temporal cortex which involves cortical regions related to functional asymmetry. The results of variation in callosal morphology are discussed as part of a possible substrate of functional asymmetry and due to variation in axonal elimination in early brain development. Sex differences were found in several aspects of callosal anatomy. (1) The difference between hand groups in the posterior body occurred in interaction with sex:handedness was a factor in callosal size in males (n = 15), but not in females (n = 35). This result is consistent with the general hypothesis of females having less clear lateralization than males. (2) Females did not have a larger overall callosum or a larger splenium, either in absolute size or size proportional to brain weight. The latter measure was considered since callosal area correlated with cerebrum weight (r = 0.48). In contrast, female of both hand groups were found to have a larger proportional isthmus compared to CRH males. (3) Of all callosal regions, only the genu and a part of the anterior body were found to be larger in absolute size in males than females. (4) Callosal size decreased with chronological age in males, but not in females.