A lesion-induced formation of an abnormal projection of hippocampal mossy fiber collaterals to the molecular layer of the fascia dentata was studied in rats. Both immature (1--30 days old) and adult rats were subjected to hippocampal and entorhinal lesions which alone or in combination removed one or more of the major afferents to the dentate molecular layer (commissural, associational, and perforant path). Some lesions in addition transected the main part of the mossy fibers en route from the dentate granule cells to the hippocampal pyramidal cells in regio inferior (CA3). The formation of aberrant mossy fiber terminals in fascia dentata (supragranular mossy fibers) was monitored by the histochemical Timm sulphide silver method, but the presence of aberrant terminals was also observed in the electron microscope. Abnormal amounts of supragranular mossy fiber terminals were found following entorhinal lesions of both immature and adult rats, but not following commissural lesions. Even larger amounts of aberrant terminals were, however, found in immature and adult rats subjected to lesions which removed most of the associational hippocampodentate projection by isolating columns of fascia dentata from major parts of the hilus (CA4). Pure transections of the fascia dentata perpendicular to its longitudinal septotemporal axis did not in itself cause aberrant supragranular terminals, although such lesions partially damaged the associational afferents. When the transections were combined with commissural lesions or entorhinal lesions or both, large amounts of supragranular terminals did, however, form at the denervated levels septal to the transection. After comparison of the amounts and distributions of the aberrant terminals found after the different lesions and in transplants of dentate tissue with different amounts of afferent input, we conclude that it is deafferentation of the dentate molecular layer, and not axotomy of the mossy fibers in the hilus of CA3 (pruning), that causes the aberrant growth of mossy fiber collaterals. Moreover, simultaneous removal of more than one afferent system seems to have a potentiating rather than a simple additive effect on the formation of supragranular mossy fibers.