Previous lesion studies have suggested that the septal-hippocampal system is involved in fear and anxiety. In this study we examined the effects on anxiety of temporary neuronal inhibition of various aspects of the septo-hippocampal system in rats. Infusions of tetrodotoxin (TTX) were used to induce reversible lesions in the fimbria fornix, medial septum, dorsal hippocampus, and ventral hippocampus. To assess anxiety we used the elevated plus-maze and the shock-probe burying tests. A reduction in anxiety in the elevated plus-maze is indicated by increased open arm exploration, whereas a reduction in anxiety in the shock-probe burying test is indicated by decreased burying behavior or increased contacts with the shock-probe. The results suggested that inhibition of the septal-hippocampal system induced site-specific anxiolytic effects that vary in nature. Tetrodotoxin lesions of the fimbria fornix increased both open arm exploration and the number of shocks taken by the rats, while having no effect on burying behavior. Both septal and ventral hippocampal lesions increased open arm exploration and decreased burying behavior, but had no effect on the number of probe shocks. Finally, TTX lesions of the dorsal hippocampus increased the number of shocks taken by the rats, but did not affect open arm activity or burying behavior. Neuroanatomical studies indicated that the effect on the number of shocks induced by dorsal hippocampal TTX lesions was not likely mediated by the amygdala. Collectively, the data suggest that the control of specific anxiety reactions is functionally segregated within different aspects of the septo-hippocampal system.