Neurons containing somatostatin (SOM) are enriched in the dentate gyrus. We sought to establish the ultrastructural localization of this peptide in the dentate gyrus of the rat brain with a double-bridged peroxidase-antiperoxidase (PAP) method localizing antisera directed against somatostatin (SOM)-28 and SOM-28. Initial light microscopic observations confirmed that the majority of perikarya and thick varicose processes with intense SOM-like immunoreactivity (SOM-LI) were observed in the hilus. Fine varicose processes with SOM-LI were found throughout all layers of the dentate gyrus but were most intense in the outer third of the molecular layer (ML), where an occasional perikaryon with SOM-LI was seen. By electron microscopy, SOM-LI was found in neuronal perikarya, dendrites, axons, and axon terminals. Two types of SOM-containing perikarya were observed. The first type was small (6-10 microns), round or avoid, and had a labeled cytoplasma with abundant Golgi complexes and a dense accumulation of PAP-reaction product. The second type of perikarya was larger (11-16 microns) and had a more abundant cytoplasm than the first type, but the Golgi complexes did not appear labeled. Most (96% of 374) of the synapses on the SOM-labeled perikarya and dendrites were from terminals without SOM-LI which formed nearly equal proportions of asymmetric and symmetric junctions. The remainder of the presynaptic terminals contained SOM-LI and made primarily symmetric synapses. Synaptic junctions from both unlabeled and labeled terminals were primarily on the shafts of the small (0.5-1.5 microns) SOM-immunoreactive dendrites. The terminals with SOM-LI (0.25-1.3 microns) contained many small, clear vesicles and from zero to four large dense-core vesicles. Terminals with SOM-LI were associated 1) with one unlabeled perikaryon or dendrite (49% of 215 in the hilus; 76% of 326 in the ML); 2) with two unlabeled perikarya or dendrites simultaneously (5% hilus; 4% ML); and 3) with one SOM-containing perikaryon or dendrite (6% hilus; 3% ML). In all three types of associations, synaptic contacts on perikarya were few while the majority were with small (distal) dendrites. Moreover, most of the terminals with SOM-LI formed symmetric junctions or lacked membrane specializations but were without any apparent glial intervention in the plane of section analyzed. The remaining SOM-labeled terminals (40% hilus; 17% ML) were without any apparent synaptic relations. However, a few of these terminals were in direct apposition to other terminals, some of which were also SOM-immunoreactive.(ABSTRACT TRUNCATED AT 400 WORDS)