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The distribution of perichromatin granules (PGs), a storage form of pre-mRNAs, was studied in supraoptic neurons of control and osmotically stimulated rats, and also after treatment with cycloheximide, a protein-synthesis inhibitor. In non-cycloheximide-treated rats, neuronal activation by dehydration significantly decreased the number of PGs. Conversely, PGs were drastically increased in the supraoptic neurons of dehydrated rats treated with cycloheximide for 4 h. This suggests that cycloheximide does not interfere with the transcriptional activation induced by dehydration, but it affects the processing of newly synthesized pre-mRNAs. Moreover, protein-synthesis inhibition was associated with the formation of intranuclear bundles of tubular filaments.

The present light and electron microscopic study deals with the morphology and organization of Cajal-Smirnow ansiform fibers (AFs) in the molecular layer of the cerebellar cortex. The cerebella of normal adult rats were processed with Cajal's reduced silver method and conventional electron microscopy. With the silver method AFs appear as isolated elements or, more frequently, as small bundles of myelinated fibers, which emerge from the medullary rays, ascend through the granular, Purkinje cell and molecular layers and curve back to reenter the granular layer or cerebellar white matter. They traced an arciform trajectory of variable width and height in the molecular layer. Relatively large bundles of AFs were rarely found. The occurrence of AFs was confirmed in semithin sections as myelinated fibers of variable diameter ranging from 1 to 6 micron. Oligodendrocytes were often observed near AFs. At the ultrastructural level, the most common type of AF is large, with a relatively thin myelin sheath and a moderately dense axoplasm. Nodal or terminal synaptic differentiations were not observed. We suggest that AFs are misoriented cerebellar mossy fibers and their occurrence may be the consequence of a small-scale error in the axonal guidance of growing mossy fibers.

The present paper describes a simple, efficient method for silver impregnation of supraoptic nuclei of the rat hypothalamus using a modification of the ammoniacal silver technique of Cajal (1903). This procedure, involving a silver-developer sequence in tissue blocks prior to plastic embedding, permits the simultaneous study of Ag-impregnated supraoptic neurons at both light and electron microscopic levels. Visualization of secretory magnocellular neurons impregnated by this technique using the electron microscope reveals a good preservation of nuclear structures. A selective accumulation of silver grains was observed over heterochromatin clumps and nucleoli, which allows the identification of the nucleolar fibrillar centers and also the dense fibrillar component as the main areas involved in the silver reaction. The meaning of such a silver-distribution pattern is discussed in the light of recent ultrastructural and biochemical data.

The present light and electron microscopic study deals with the morphology and staining properties of two intranuclear inclusions - the "accessory body" of Cajal and the "coiled body"--in the supraoptic nuclei of adult rat hypothalamus, and supports the assumption that these structures represent the same intrinsic component of the neuronal nucleus. Consequently, we propose to term it "accessory body". The structure of this body was visualized by several different staining procedures: conventional electron microscopic techniques, a silver reaction, and the regressive EDTA staining for ribonucleoproteins. The silver-impregnation method employed here, which consists of a silver development sequence on hypothalamic tissue blocks prior to plastic embedding, permitted the study of supraoptic neurons at both light and electron microscopic levels. The nature and origin of "accessory bodies" are suggested and their possible functional role is briefly discussed.

Cilia 9+0 have been found in differentiating neurosecretory neurons of the hypothalamic supraoptic nuclei of Wistar rats. These ciliated neurosecretory cells were always observed during the neonatal period of the rat and no more than one cilium per cell has been encountered. Single cilia arising from typical basal bodies were usually located in deep invaginations of the neuronal perikaryon, but can also be seen in superficial positions. Their basal bodies were commonly found in the vicinity of Golgi complexes, and associated structures such as striated rootlets and alar sheets were also present. In addition, single centrioles frequently appeared in these neurons, occurring close to basal bodies but also in centrosomatic areas. The respective roles of these morphological features are suggested and their possible ontogenetic significance is briefly discussed.

Ultrastructural study of the hypothalamic ventromedial nucleus during the postnatal period showed the presence of nuclear inclusions in immature glial cells. These inclusions, identified as "coiled bodies", consist of round-to-oval formations of coiled electron-dense strands embedded in a less dense fibrillar matrix. Coiled bodies are located free within the nucleoplasm, showing no specific relationships with the nucleolus or the nuclear membrane. The cells containing coiled bodies were typified as oligodendrocyte precursors, mainly oligodendroblasts. The coiled bodies were not found in mature glial cells nor in other types of immature glial elements. The nature and possible functional role of coiled bodies are suggested in the light of recent morphological and biochemical data.

The morphological study of the rat fastigial nucleus with the Golgi-Rio Hortega method showed the presence of glial perineuronal nets surrounding the large neurons, but not the small ones. This perineuronal net appeared as a mesh of tenuous glial processes which covers the neuronal perikarya and proximal dendrites. The small alveolate compartments in this mesh seem to correspond to the holes for the synaptic boutons. Our results also indicate that the perineuronal net is derived from interneuronal protoplasmic and velate astrocytes. Using camera lucida drawings of this perineuronal net we have made a quantitative estimation of the size and density of synaptic boutons on these large neurons. The average numerical density of synaptic boutons was about 19 per 100 micron 2 of the neuronal surface, the mean area of the synaptic holes being 2.5 micron 2. Furthermore, the quantitative data evidence that about 52.5% of the neuronal surface is presumably occupied by synaptic boutons whereas the remaining 47.5% is covered by the glial processes of the perineuronal net. Semithin sections prepared from thick Golgi sections were used for the cytological study of the neurons surrounded by this glial pericellular network. The possible functional significance of the perineuronal net in the regulation of synaptic transmission in the fastigial cerebellar nucleus is briefly discussed.