The hypotheses that the medial septal area (MSA) is critical for working memory and that MSA neural activity is positively regulated by cholinergic inputs leads to two testable predictions: (1) working memory can be bidirectionally modulated by muscarinic manipulations of the MSA and (2) muscarinic activation of the MSA can enhance memory under conditions of mnemonic impairment. Memory was assessed by T-maze spatial alternation following intraseptal infusion of muscarinic drugs in rats pretreated with intraperitoneal (IP-) injections of scopolamine. Scopolamine dose-dependently impaired working memory and shifted the hippocampal theta activity to a higher peak frequency. Intraseptal scopolamine mimicked the behavioral effects of IP-scopolamine, and intraseptal carbachol appeared to reverse both the behavioral and physiological effects of IP-scopolamine. The results indicate that the amnestic effect of antimuscarinic drugs may be due to an interaction in the MSA and that conditions of memory impairment may be alleviated by selective muscarinic activation of the MSA.