Relationships between glial and neuronal elements in the development of long term cultures of the spinal cord of the fetal mouse.
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Electron microscopic and Golgi studies on the inferior lobes of sunfish and goldfish are described. The inferior lobe consists primarily of a nucleus ventricularis of densely packed cells surrounding the lateral recess of the third ventricle, and a peripherally situated nucleus diffusus consisting mostly of scattered neurons. A cell-sparse zone of dense neuropil is located between the two cellular areas. Neurons of both nuclei have spiny dendrites and axons which originate from basal dendrites. In some cases axons are found to send a collateral into the cell-sparse zone. Neurons of the nucleus diffusus possess collaterals that extend a considerable distance within the nucleus itself. The ultrastructure of cells of both nuclei reveals cytoplasmic organelles typical of most neurons. Synapses containing dense-cored and clear vesicles are present on the spines and shafts of the dendrites of both neuronal types. In only rare cases synapses were observed on the soma of neurons of the nucleus ventricularis. Possible anatomical substrates involved in the control of feeding and aggression in teleosts are considered in light of the present findings. Morphological similarities of the inferior lobes and related areas in various fishes and amphibians are discussed and their possible significance for the understanding of the evolution of hypothalamic mechanisms is considered.
The ultrastructure of axons in the preganglionic cervical sympathetic trunk of the mouse is described with emphasis on the number, distribution and stability of fibrous elements in the axoplasm. Neurofilaments outnumbered microtubules in myelinated and non-myelinated axons of all sizes, and the ratio of neurofilaments to microtubules in non-myelinated axons at each point studied was fairly consistent and independent of axonal diameter. The density of neurofilaments and microtubules, however, was greater in axons of progressively smaller diameter. In non-myelinated axons and small myelinated axons neurofilaments were uniformly distributed throughout the axoplasm resulting in minimum and maximum interfilament distances of 300 angstrom and 500 angstrom respectively; the spacing of fibrous elements within any one axon was dependent upon its diameter and position with respect to the superior cervical ganglion in the preganglionic trunk. The maximum interfilament distance was also found in large myelinated axons where neurofilaments, occurring in fascicles, were separated by distances of approximately 500 angstroms. Cytochemical staining of axons with lanthanum hydroxide, ruthenium red or alkaline bismuth delineated the delicate filamentous matrix interconnecting microtubules, neurofilaments and other organelles in the axoplasm. Alkaline bismuth stain was most intense in myelinated axons where heaviest deposition of reaction product was associated with neurofilaments. Treatment in vitro of the cervical sympathetic trunk with 5 X 10(-5) M vinblastine sulfate dissociated microtubules and induced formation of crystalline arrays of "tubular" elements. A uniform center to center spacing of 250-300 angstrom was found for crystalloids in non-myelinted axons; however, in myelinated axons the center to center spacing was not uniform and varied in the range 300-600 angstrom. Neurofilaments and their surface projections were unaffected by vinblastine. Fixation in the presence of lanthanum enhanced delineation of crystalloid elements. Exposure of 0-4 degrees C for up to three hours had no consistent effect on microtubules or neurofilaments. In contrast, cold treatment disrupted the delicate axonal matrix and resulted in the formation of aggregates of coarse flocculent material in the axoplasm.
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Antisera to the 150K-dalton polypeptide of the bovine neurofilament triplet and chicken neurofilament antisera reacting with the 70K protein in isolated bovine brain filaments stained the same structures in rat cerebellum by immunofluorescence and peroxidase-antiperoxidase methods, that is Purkinje cell baskets, thin nerve fibers in the lower half of the molecular layer and myelinated axons. The 150K bovine neurofilament antisera did not stain large motor neurons in the anterior horns of the spinal cord in rat and rabbit, nor aluminum-induced neurofibrillary tangles in the rabbit. All these structures were demonstrated by the chicken neurofilament antisera and by silver neurofibrillary methods. IDPN-induced axonal balloons containing accumulations of neurofilaments were equally well stained by bovine 150K and chicken neurofilament antisera. These data suggest that the 150K polypeptide of the neurofilament triplet is not a subunit of the neurofilament core and probably plays a role in axonal transport.
In rabbit retina, neurofibrillar methods stain two populations of amacrines whose cell before are located on either side of the inner plexiform layer: one in the ganglion cell layer and the other at the inner margin of the inner nuclear layer. The stained amacrines in the ganglion cell layer have the distinctive cytology of "coronate" amacrines described from Nissl-stained retina (Vaney, '80a) and account from about 85% of the displaced amacrines in the rabbit retina. The coronate amacrines have a streak topography similar to that of the ganglion cells; they comprise about 32% of the neurons in the ganglion cells layer although their proportion increases with eccentricity from the visual streak. The cytology of the neurofibrillar stained amacrines in the inner nuclear layer resembles that of the displaced amacrines and their densities are almost equal. The cell bodies of the stained amacrines are smaller than those of the displaced amacrines but larger than most others in the inner nuclear layer; they account for some 2% of the neurons in the amacrine sublayer. Although the cell bodies of both populations are distributed rather evenly, the mosaic of the inner nuclear layer cells is more regular than that of the ganglion layer cells. We propose that the two populations of amacrines stained by neurofibrillar methods correspond to the acetylcholine synthesizing cells labelled by Masland and Mills ('79).