Vasoactive intestinal polypeptide-immunoreactive interneurons have been classified according to their axonal and dendritic patterns and neurochemical features in the hippocampus of the rat. A correlation of these characteristics unravelled three distinct types of vasoactive intestinal polypeptide-containing cells. Interneurons forming a dense axonal plexus at the border of stratum oriens and alveus always contain the calcium binding protein, calretinin, but lack the neuropeptide cholecystokinin. The axon of another type of vasoactive intestinal polypeptide-positive interneuron surrounds pyramidal cell bodies in a basket-like manner, and co-localizes cholecystokinin but not calretinin. Vasoactive intestinal polypeptide-containing cells projecting to stratum radiatum form two subsets distinguished by dendritic morphology. Those with dendrites restricted to stratum lacunosum-molecular lack both calretinin and cholecystokinin, whereas the other subtype with dendrites spanning all layers contains calretinin in 40% of the cases and occasionally also cholecystokin. GABA was shown to be present, and the calcium binding proteins calbindin D-28k and parvalbumin absent from all three types of vasoactive intestinal polypeptide-positive interneurons. The specific dendritic and axonal arbours imply different input and output properties for the three interneuron types. The correlation of these features with the content of neurochemical markers strongly suggests that they are specialized for distinct inhibitory functions in the hippocampal network.