Male Long-Evans rats received micro-injections of either N-methyl-D-aspartate (NMDA) in the medial septum/vertical diagonal band (MS/DB), 5,7-dihyroxytryptamine (5,7-DHT) in the fimbria/fornix and cingulate bundle or combined NMDA/5,7-DHT micro-injections. NMDA administration caused considerable damage to the MS and enlarged the lateral ventricles. It reduced the activity of choline acetyltransferase as well as the intensity of acetylcholinesterase staining in the hippocampus. 5,7-DHT selectively reduced the concentration of hippocampal serotonin. The rats were assessed for spatial memory in the Morris water maze and the radial arm maze (reference and working memory version). The 5,7-DHT-induced lesion of hippocampal serotonin had no effect by itself on either task. However, it augmented the reference memory impairment caused by the NMDA-induced lesion and delayed the recovery from NMDA-induced impairment of working memory on the radial maze. Combined damage of hippocampal cholinergic and serotonergic afferents did not severely affect spatial memory.