The serotoninergic nerve cell body population of nucleus raphe dorsalis (RD) was identified by radioautography following cerebroventricular instillation of tritiated serotonin ([3H]5-HT) in adult rats pretreated with a monoamine oxidase inhibitor. Series of histological sections taken throughout the midbrain and upper pons exhibited a similar distribution and number of labeled nerve cell bodies in RD after prolonged administration of either 10-5 or 10-4M [3H]5-HT or 10-4M [3H]5-HT and 10-3M nonradioactive noradrenaline. This allowed systematic mapping and quantification of serotoninergic nerve cell bodies at various levels of the RD. Their extrapolated total number averaged 11,500. Twice as many unreactive (nonserotoninergic) neurons were present within the same region. In electron microscope radioautographs, the labeled cells were usually larger (17.9 micrometer mean diameter) than their unlabeled congeners (13.1 micrometer), but stereological sampling of their perikarial organelle content failed to reveal any difference in cytoplasmic composition. Few [3H]5-HT-labeled axonal varicosities were observed in RD and none were found in close apposition or in synaptic junction with labeled nerve cell bodies, dendrites, or unreactive perikarya. A detailed statistical analysis of silver grain distribution in both labeled and "unlabeled" nerve cell bodies, indicated that in the former, but not in the latter, dense bodies had a relatively high affinity for [3H]5-HT. Mitochondria and the cytoplasmic membrane were the only other organelles to show higher labeling indices in labeled than in unlabeled cells. Other sites of [3H]5-HT localization could be ascribed to artefactitious cross-linkage of the tracer by the fixative, since they had the same relative affinity in the two cell populations. These results provide new insights into the morphology and cytofunctional properties of the 5-HT neurons of rat RD.
In a well-defined sector of adult rat hippocampus (CA1, stratum radiatum), the ultrastructural features of acetylcholine (ACh), noradrenaline (NA), serotonin (5-HT) and GABA axon terminals (varicosities) were compared by electron microscopy after immunostaining with antibodies against choline acetyltransferase, NA, 5-HT and glutamic acid decarboxylase. Approximately 100 sectional profiles of each type were analyzed for size, presence of a synaptic membrane specialization (synaptic incidence) and composition of the microenvironment. An equivalent number of immunonegative varicosity profiles selected at random from the same micrographs were similarly examined. ACh, NA and 5-HT varicosity profiles were of comparable size, and significantly smaller than GABA profiles. They exhibited a low frequency of junctional specialization, amounting to 7%, 15% and 21%, respectively, when extrapolated to the whole volume of these terminals. In contrast, GABA varicosities appeared entirely synaptic. The ACh, NA and 5-HT varicosities also differed from their GABA counterparts in being juxtaposed to a greater number of unlabeled axonal varicosities and a lower number of dendritic branches. In addition, the microenvironment of immunostained terminals showed a much lower number of dendritic spines than that of immunonegative varicosities. This latter finding was viewed as another indication that predominantly asynaptic varicosities do not maintain particular relationships with their immediate surround. It was also concluded that volume transmission represents a major mode of transmission for ACh, NA and 5-HT in adult rat hippocampus, thus contributing to the properties and functions assigned to these transmitters in this part of brain.
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