Sex and species differences in mouse and rat forebrain commissures depend on the method of adjusting for brain size

Sex differences in the forebrain commissures (corpus callosum, hippocampal commissure, and anterior commissure) were examined in B6D2F2 hybrid mice and Sprague-Dawley rats. Twenty-four male-female littermate pairs of mice were perfused at each of 21, 42 and 63 days of age and the midsagittal area of the commissures was measured from en bloc stained tissue. Twenty-two male-female littermate pairs of rats were examined at 110 days of age using the same methods. Male mice had larger bodies than females but no sex differences were found for mouse brain weight or commissure areas. In contrast, a significant sex difference was found for rat body, brain, corpus callosum and hippocampal commissure sizes. Four methods were used to adjust for differences in brain size (ratio, geometric, linear regression, and allometric). When the two species were analysed separately, neither mice nor rats showed significant sex differences in commissure areas relative to brain size if regression or allometric adjustments were made. Even when data from mice and rats were combined into one large group with a wide range of values, no species or sex differences were apparent after adjustments were made for brain size with either the regression or allometric methods. The use of ratios to adjust for differences in overall size is not recommended, especially because this method does not effectively remove the influence of brain size from commissure size; a substantial correlation is often present between the ratio and brain size.