Serotonergic ependymal fibres in rat and monkey: light and electron microscopic immunocytochemical study

The distribution of serotonin (5HT) immunoreactive fibres in the ependymal epithelium of aqueductus cerebri in adult rat and monkey was studied by means of immunocytochemical methods using specific antibodies against 5HT. Light microscopic examination of immunostained coronal sections of mesencephalon showed, in tryptophan and pargylin pretreated animals, abundant serotonergic fibres distributed along the ependymal cells of the aqueduct, forming supraependymal and subependymal plexi. Serotonin fibres lying either supraependymally or ending freely in the aqueduct lumen contributed to the formation of a rich 5HT containing network on the ependymal surface. Electron microscope images showed dense 5HT-immunoreactive (5HT-IR) profiles with ultrastructural characteristics of axon terminals ending on the ependymal cells. Dense diaminobenzidine (DAB) deposits were found in the axoplasm, on outer mitochondrial surface and in vesicles. No synaptic contacts were observed between 5HT-IR terminals and ependymal cells. Unstained microvilli and cilia were also observed in the aqueduct lumen. Serotonin immunoreactivity disappeared from ependymal fibres in animals treated with parachlorophenylalanine, an inhibitor of 5HT synthesis. 5HT containing fibres described in this paper may be the source of 5HT and its metabolite 5-hydroxyindoleacetic acid (5HIAA) found in cerebrospinal fluid (CSF) and of clinical relevance in some psychiatric conditions such as depression, suicidal attempts, etc

Distribution of GABA in the golden hamster retina: An immunocytochemical study

Distribution of the inhibitory neurotransmitter GABA was studied in the golden hamster retina using immunocytochemistry at cellular and subcellular levels. GABA-immunoreactivity was observed in somata of amacrine, displaced amacrine and horizontal cells. GABA immunoreactive fibers were abundant in the inner plexiform layer. Ultrastructural analysis exhibited dense GABA-immunoreactive deposits in amacrine cell somata, processes and terminals. Immunolabelling was also observed in the cytoplasm of horizontal or interplexiform cells and displaced amacrine cells. In every case DAB deposits were observed in the cytosolic compartment, attached to the inner surface of cell membranes and to outer mitochondrial membranes. Immunolabeled terminals predominated in the inner plexiform layer and immunoprecipitates were also observed attached to the outer face of vesicle membranes as well as completely filling synaptic vesicles. Both clear and dense core vesicles were observed. The present results are similar to those obtained in other mammalian species showing GABA immunoreactivity in amacrine, displaced amacrine and horizontal cells

Ultrastructural study of reaggregated culture of brainstem: synaptogenesis

The synaptogenesis and the morphological differentiation of neural cells were studied in aggregating cultures. Brainstems of 14-15 days old rat embryos were removed and the area located between the mesencephalic flexure and the caudal portion of metencephalon was dissected and mechanically dissociated to single cells. These cells reassociated forming highly organized aggregates in which differentiation took place. Samples were harvested after different time periods, fixed and processed for electron-microscopic study. After one day in culture the aggregates were composed by rounded undifferentiated cells. These cells had a high nuclear/cytoplasmic relation, were devoid of processes and were separated by great intercellular spaces. At the end of the first week of culture cell differentiation and extension of processes were evident. A loose neuropil appeared: it was composed by abundant growing neurites and growth cones. Later, the neuropil became more compact and glial processes and synaptic terminals filled with vesicles appeared. The early appearance of vesicles in the synaptic endings was the first evidence of synaptogenesis. Post and presynaptic membrane densities appeared later, and fully mature synaptic contacts were seen by the end of the 3rd week in culture. Scarce myelin sheaths were observed after 35 days in vitro

Immunocytochemical localization of sorotonin in the pineal gland

Se estudió la localización de serotonina (5HT) en la glándula pineal de ratas en condiciones de luz diurna por medio de microscopía óptica y electrónica. Se usó un anticuerpo policlonal para 5 HT y el método inmunocitoquímico de peroxidasa antiperoxidas (PAP). La glándula mostró fibras varicosas inmunomarcadas distribuidas por toda su extensión. Las observaciones ultraestructurales revelaron la presencia de depósitos electrodensos de diaminobencidina (DAB) en terminales localizados principalmente en los espacios perivasculares