Significance The relation between structural and functional connectivity has profound implications for our understanding of cerebral physiology and cognitive neuroscience. Yet, this relation remains incompletely understood. Cases in which the corpus callosum is sectioned for medical reasons provide a unique opportunity to study this question. We report functional connectivity assessed before and after surgical section of the corpus callosum, including multiyear follow-up in a limited subsample. Our results demonstrate a causal role for the corpus callosum in maintaining functional connectivity between the hemispheres. Additionally, comparison of results obtained in complete vs. partial callosotomy demonstrate that polysynaptic connections also play a role in maintaining interhemispheric functional connectivity.