The light and electron microscopic localization of antigenic sites for a polyclonal antiserum directed against the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH), was examined in the hippocampal formation of the rat brain with a double-bridged peroxidase-antiperoxidase method. By light microscopy, the majority of varicose processes with intense TH-like immunoreactivity (LI) were contained in the hilus of the dentate gyrus (DG) and strata radiatum and lacunosum-moleculare of the CA3 region of the hippocampus. Only a few immunoreactive fibers were observed in the molecular and granule cell layers of the DG, in strata oriens and pyramidale of CA3, and in all layers of CA1. Electron microscopy confirmed that these labeled processes were primarily axons and axon terminals. Terminals with TH-LI were 0.4-1.1 micron in diameter and contained many small clear vesicles and from 0 to 3 larger dense-core vesicles. The number and types of associations formed by terminals with TH-LI were remarkably similar in the DG and hippocampus proper despite known differences in intrinsic cells and function. In both regions, the majority of terminals with TH-LI formed junctions on small (distal dendrites (52% of 112 in the DG; 67% of 116 in CA3) and dendritic spines (30% in the DG; 18% in CA3) that were both asymmetric and symmetric. In the DG, axosomatic junctions (2% of 112) were symmetric and occurred exclusively on the perikarya of granule cells, whereas junctions on large (proximal) dendrites were more numerous (16%), exhibited symmetric as well as asymmetric membrane specializations, and were of both granule (molecular layer) and nongranule (hilus) cell origin. In CA3, synaptic contacts on perikarya (5% of 116) and large (proximal) dendrites (10%) of both pyramidal cell and nonpyramidal cell origin were few and all symmetric. The distribution and types of synaptic associations formed by terminals with TH-LI in the CA1 region paralleled that seen in the CA3 region. In both the dentate and hippocampus proper, 10% of the terminals with TH-LI were observed closely apposed to unlabeled terminals that formed asymmetric synapses with dendrites and dendritic spines. In rare instances, TH-immunoreactive terminals were found in close association with the basement membrane of blood vessels, astrocytic processes, or with other unlabeled terminals not forming recognizable junctions. In addition TH-LI was occasionally detected within the cytoplasm of a minority of astrocytes.(ABSTRACT TRUNCATED AT 400 WORDS)
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)
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