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The mesencephalic neural plate of early-somite-stage mouse embryos differentiated underneath the renal capsule to form mostly neural tissues together with other tissues some of which were probably of neural crest cell origin. The capacity to form non-neural tissues such as skeletal tissues and melanocytes was lost at about the 5-somite stage. The lateral areas of the plate tended to form non-neural tissues more than the medial areas. The cephalic neural plate of presomite head-fold-stage embryos differentiated extensively to form both ectodermal and mesodermal tissues. However, upon completion of neurulation, the mesencephalic neuroepithelium of forelimb-bud-stage embryos gave rise to neural tissues only. Therefore there is a progressive restriction in the histogenetic capacity of the mesencephalic neural plate during neurulation and this could be attributed to the cellular commitment for neural differentiation and the loss of the neural crest cells.

Three muscle fibers were identified by electron microscopy within a nerve of a frog muscle. They resembled extrafusal muscle fibers but were located in an endoneurial rather than in an endomysial compartment. To call these endoneurial muscle fibers the obvious continuation of extrafusal fibers of a muscle spindle is certainly unwarranted; to label these fibers ectopic and to let the matter rest there is probably an understatement of sorts.

Monoclonal antibodies to squid neurofilament (aNFP) and intermediate filament (aIFA) proteins were used as probes for the biochemical and immunocytochemical analyses of neurofilament structure and distribution in the squid giant axon and stellate ganglion. On Western blots the aNFP antibody stained exclusively the 220 kDa and high-molecular-weight (HMW) components of neurofilaments in the giant axon, whereas the aIFA antibody primarily labeled the 60 kDa protein in the giant axon and the 60 and 65 kDa proteins in the stellate ganglion. Dephosphorylation of axoplasmic proteins by alkaline phosphatase resulted in a decrease in the molecular weights of both the 220 kDa and HMW neurofilament proteins and a concomitant loss of reactivity with the aNFP antibody on Western blots. This indicated that the aNFP antibody is specific for a phosphorylated epitope in the neurofilament. Increased dephosphorylation of the 220 kDa protein led to an enhanced immunostaining of the resultant 190 kDa polypeptide by the aIFA antibody, suggesting that the phosphorylated epitope may mask the conserved epitope recognized by aIFA. Light and electron microscopic immunocytochemical studies show intense labeling by the aNFP antibody in the giant axon. In contrast, the aIFA antibody labeled the glial cells around the giant axon intensely, while labeling of the giant axon itself was considerably less than that with the aNFP antibody. Since the 60 kDa protein in axoplasm is intensely stained by the aIFA antibody on Western blots, the relatively low amounts of labeling seen on semithin and thin sections of the giant axon by this antibody may be due to the masking of the 60 kDa protein by in situ fixed axoplasmic proteins. However, the aIFA antibody intensely labeled glial cells within the stellate ganglion and "islands" of filaments and nuclear membranes within ganglion cells. No reactivity for either antibody was seen in synapses. The aNFP antibody specifically labeled "beadlike" portions and cross-bridges on the axonal neurofilaments, suggesting that these components consist of the 220 kDa and HMW proteins. In contrast, the aIFA antibody labeled relatively smooth filaments in ganglion and glial cells. These data suggest that the 65 kDa protein represents the squid glial filament protein and that the 60 kDa protein found in axoplasm represents the low-molecular-weight subunit in the axonal neurofilament. The latter appears to be formed and/or organized in "islands" of filaments within ganglion cells.(ABSTRACT TRUNCATED AT 400 WORDS)

The ganglion of Cephalodiscus gracilis M'Intosh 1882 is entirely intraepithelial and located in the dorsal epidermis immediately behind the tentacular apparatus that is formed by the mesosome (collar). A characteristic feature of the ganglion is a well-developed neuropile in which different types of nerve fibres can be discerned, many of which contain small granules with electron-dense contents. There are no glia-like cells in association with these fibres. Only slender basal processes of epidermal epithelial cells traverse the neuropile. In the depth of the epithelium the neuropile borders the epidermal basal lamina; apically it is covered by a layer of cell bodies, the majority of which belong to what appear to be ordinary ciliated epidermal cells. Besides these epidermal cells the perikarya of two additional types of cells, which are considered to be neurons, can be discerned. One type is characterised by many rough endoplasmic reticulum cisterns and mitochondria, the other by abundant small, electron-dense granules. The nuclei of these cells are comparatively pale and contain a prominent nucleolus. The neuron cell bodies do not form a distinct layer; but they are loosely distributed somewhat deeper than those of the ordinary epidermal cells. They probably send off an apical process to the epidermal surface and a basally directed one into the neuropile. The ganglion has been compared to the nervous systems in cnidarians, some spiralians, and especially other hemichordates, echinoderms, and chordates; it is found to be of primitive rather than degenerate nature. Furthermore, the possible functional significance of its close connection to the food-capturing tentacular apparatus is discussed.

The cytoskeleton of a newly isolated mouse embryo brain-derived cell line and of dissected mesencephalon-rhombencephalon samples of the 10- to 11-day-old mouse embryo, constituted of a ventricular zone only, has been examined by immunocytochemistry, electrophoretic separation and Western blotting techniques. In accordance with their epithelial organization, the ventricular cells contain cytokeratins as main constituents of their cytoskeleton. Vimentin has been also revealed as early as day 10.5 of gestation. The complex cytokeratin polypeptide pattern puts this histological type of epithelia into the category of complex, rather than simple, epithelia and calls for explanations other than keratinization for the presence and functional role of the high-molecular weight, basic keratins. The cytoskeletal composition of the embryo brain-derived gliogenic cell line reflects the vimentin- and cytokeratin-positive character of the ventricular cells.

All neuronal cells in the dwarf male of the dimorphic polychaete species Dinophilus gyrociliatus were individually identified by means of serial ultrathin sections. Altogether 68 neural cells--including 40 sensory neurons and 2 glial cells--constitute a small but complex nervous system. Fifty-three neural cells are located in three pairs of ganglia and connected by paired nerve cords. The prominent frontal ganglia, each consisting of a well-developed neuropile and surrounded by 20 or 21 neural cells, represent the animal's brain. The ventral ganglia contain only 2 neurons each. The penis ganglia--four cells each--are associated with the copulatory organ. A conspicuous circumpenial fiber mass surrounds the basal part of the penis. The effector cells--22 multiciliated epidermal cells, 34 muscle cells, and different gland cells (?)--were also reconstructed and their innervation was partly elucidated. Sensory-motor neurons were unambiguously identified. They are discussed in regard to the small body size of the animal. The male's nervous organization resembles a very simple rope ladder and may represent a reduced derivative of a nervous system in normal-sized males of monomorphic species. Similarities, however, also occur with the developing nervous system of a planktotrophic metatrochophore. The neuronal organization, with its two centers (frontal ganglia and ventral ganglia vs. penis ganglia and circumpenial fiber mass), accords well with the bipartite behavioral pattern, which is entirely devoted to locomotion and copulation, respectively.

Electron microscopic examination of the developing neural tube in 11th to 13th day rat fetuses revealed a new cell surface relationship between differentiating neuroepithelial cells. Cytoplasmic projections possessing terminal dilations were observed extending from neuroepithelial cells through cytoplasmic furrows into large coated pits at the surface of adjacent cells. This cell-to-cell relationship provides a mechanism for the internalization of surface molecules and possibly even cytoplasmic constituents. Communication between donor and recipient cells mediated in this way suggests a route for the sharing of macromolecules, including cytoplasmic fragments, which could function as regulators in embryonic development and differentiation.

The nervous system (NS) of Microstomum lineare (Turbellaria, Macrostomida) was studied by electron and light microscopy, combined with fluorescence histochemistry (Falck-Hillarp method for biogenic monoamines). The NS is primitively organized, with a bilobed brain, two lateral nerve cords lacking commissures, and peripheral nerve cells scattered along the nerve cords. The stomatogastric NS, with a pharyngeal nerve ring, is joined to the central NS by a pair of connective ganglia. A green fluorescence in all parts of the NS indicates catecholaminergic neurons as the dominant neuron type. Ultrastructurally, two types of neurons were identified on the basis of their vesicle content: 1. Aminergic (catecholaminergic) neurons containing dense-core vesicles of varying electron-density and size, i.e., small dense-core vesicles (diameter 50--100 nm), vesicles with a highly electron-dense core (60--140 nm), and vesicles with an eccentric dense-core. 2. Presumed peptidergic neurosecretory neurons containing large granular vesicles (diameter about 200 nm) in the stomatogastric NS and peripheral parts of the central NS. In light microscopy, paraldehyde-thionin stained neurons were observed in the same areas.

The structure and distribution of synapses on nonspiking local interneurones in the metathoracic ganglion of the locust was revealed by electron microscopy following intracellular injection of horseradish peroxidase (HRP). Before staining, each interneurone was characterized physiologically as nonspiking and its output effects on motor neurones innervating muscles in a hindleg were investigated. Three nonspiking interneurones of different morphologies, each typical of a previously described population, were selected for detailed study. The first has a dorsal soma and ipsilateral neuropilar branches, the second a ventral soma and ipsilateral branches, and the third a ventral soma and contralateral branches. The somata have few trophospongial invaginations, and most of their volume is occupied by the nucleus. The initial parts of the primary neurites are either wrapped in glia or isolated in tracts from the neuropile and thus do not participate in synaptic interactions. Some of the larger secondary neurites are also wrapped in glia, but others both make and receive synaptic contacts. Output synapses have an array of some 500-1,600 round, agranular vesicles (diameter 47.0 +/- 5.7 nm; mean +/- S.D., n = 97) associated with a bar-shaped presynaptic density up to 0.3 micron long. Two postsynaptic processes, whose diameter can vary greatly, are usually associated with each presynaptic density. Processes making input synapses onto nonspiking local interneurones typically contain round, agranular vesicles and often make several contacts within a few microns. Serial reconstructions from one of the interneurones revealed input and output synapses intermingled on the larger processes with outputs dominating by a factor of 3:1, whereas on some of the thinner processes only input synapses are present. In the other two interneurones, however, both input and output synapses are present on the fine branches. No feature of the structure or distribution of synapses observed here on the nonspiking local interneurones distinguishes them from spiking neurones in the same ganglia.

The number of fibers was analysed on cross section electron micrographs (x 1.000 and 4.500). About 300 axons of motoneurons leave the Ist thoracic ganglion. Approximately 180.000 sensory fibers enter the ganglion by lateral nerves. More than 90% of these fibers terminate on the ganglion neurons. The ganglion has descending connections with more caudally lying portions of the cord. The suboesophageal ganglion is closely associated with the 1st thoracic and other ganglia, and presumably contains a large vegetative center. The brain receives considerable information from the body via primary as well as secondary fibers, and controls the activity of the nerve cord through the system of various descending connective fibers.

Cyclic phenyl saligenin phosphate (PSP) proved to be a potent delayed neurotoxin, eliciting clinical disease and lesions, and depressing neuropathy target esterase and plasma cholinesterase at much lower doses than the protoxicant tri-ortho-tolyl phosphate (TOTP). Using adult White Leghorn chickens, we noted qualitative similarities in clinical signs and peripheral nerve and spinal cord lesions elicited by PSP and the TOTP. Ataxia and weakness were prominent clinical effects. Lesions began as a distal axonopathy affecting larger myelinated fibers in spinal cord white matter and peripheral nerve. The latter were studied in detail. Major features of the lesion were intra-axonal collections of mitochondria, dense and lamellar bodies, and granular degeneration of neurofilaments. These led to Wallerian-like degeneration. Percentages of teased peripheral nerve fibers demonstrating such degeneration correlated with severity of clinical signs.

Electron microscopic observations have been made on transverse sections of the auditory (tympanal) nerve of the crickets at various levels from the tympanal ganglion (close to the ganglion, 150, 250, 1,000, 2,000 and 3,000 mu from the latter). In the vicinity of the ganglion, axons of the auditory receptors are separated from each other by the processes of Schwann cells. Beginning from the level of 150-250 mu, the axons are subdivided and form, with the help of collaterals, a subreceptor plexus which is similar to that found in the auditory system of the locust. Collaterals of an axon pass to the adjacent axons and deeply penetrate into the latter loosing their Schwann sheath, so that axonal membranes form a direct contact. At the site of these contacts, intracellular cleft is equal to 50-80 A. No synaptic vesicles were found in the region of the plexus. At the level of 3,000 mu, the auditory axons again attain a roundlike form and become completely isolated from each other by the processes of Schwann cells. It is suggested that in the region of the subreceptor plexus, electrotonic interaction between the receptors of the crickets takes place, as it is common in the auditory sistem of insects.

Hyaluronic acid (HA) added to the medium of quail neural tubes explanted in vitro influences the number of migratory neural crest cells that emerge, compared with controls. Neural crest cells were counted with an ocular grid after 20 h of migration into 0.1 mm wide areas or 'bins' lying parallel to the neural tube, and the results were analyzed by linear regression. A low concentration of HA (5 micrograms/ml) significantly decreased the total number of neural crest cells in all bins adjacent to the neural tube, whereas several high concentrations of HA (250, 500, and 1000 micrograms/ml) significantly increased the number of neural crest cells. Intermediate concentrations of HA (50 and 100 micrograms/ml) did not differ from that of controls. Linear regressions of number of cells versus distance from the tube showed no significant differences among the slopes of control, low HA, and high HA treatments, providing evidence that HA does not influence the rate of cell migration. Scanning electron microscopy showed that cells in neuroepithelia exposed to low HA (5 micrograms/ml) appeared in tighter contact, while cells of neuroepithelia in high HA (500 micrograms/ml) appeared more loosely organized, compared with controls. Cells in tight contact could be restrained from leaving the neuroepithelium, whereas cells in loose contact could more readily move out of the neural tube, thus explaining the differences in cell numbers in low HA and high HA, respectively. We conclude that HA can be a factor in the differential adhesivity among neuroepithelial cells and may be important in the initial separation of the neural crest from the neural tube.

To investigate the calcium requirement for the maintenance of the elevation of the neural folds in mammalian embryos 10.4-day-old-rat embryos, at a stage when the cephalic neural folds had elevated but not yet fused, were cultured in salines containing different concentrations of calcium. At 3 x 10(-3) M calcium the neural folds remained intact. As the concentration of calcium was reduced the elevated cephalic neural folds re-opened, flattened and then, at 10(-5) M calcium, collapsed. Replacement of calcium by strontium at 5 x 10(-3) M did not prevent the collapse of the cephalic neural folds, although at higher concentrations strontium partially prevented the collapse of the neural folds. In the cervical region most of the neuroepithelial cells were apically tapered. In intact cephalic neural folds nearly half of the cells were apically tapered. In cephalic neural folds which had been induced to collapse by culture in saline with low calcium there was an increase in the proportion of round cells and a decrease in the proportion of apically tapered cells.

The development of the cutaneous pectoris muscle was studied and compared with the differentiation of its peripheral nerve in bullfrog (Rana catesbeiana) tadpoles and frogs by light and electron microscopic techniques. This muscle preparation was chosen for this study because it possesses a number of advantages for (and has become a model system for) the study of correlated nerve-muscle development. At the earliest stage examined (stage XI) the presumptive muscle did not contain any contractile or morphologically distinguishable myotubes, but was contacted by the well-defined cutaneous pectoris nerve trunk. Myotubes were present at stage XII, the same time that nerve-associated acetylcholine receptor aggregations and nerve-evoked muscle contractions were first observed. The adult number of axons was present in the cutaneous pectoris nerve at stage XII, but no axons were myelinated. Gradually thereafter, the number of muscle fibers increased and the cutaneous pectoris axons became myelinated. By stages XX and XXI, but prior to metamorphic climax (stage XXV), the adult numbers of cutaneous pectoris muscle fibers and myelinated and unmyelinated nerve fibers were present. These numbers did not change significantly between stages XX and XXI, through metamorphosis, and in the adult, even during the period of the most rapid loss of multiple innervation in the first 2 weeks after metamorphosis. These results show that the nerve was present and in contact with the cutaneous pectoris muscle from the earliest stages of development prior to muscle differentiation, at a time when the muscle was a disorganized mass of undifferentiated cells. Such early contact suggests that the nerve may have a significant influence on muscle maturation.

The brains of individuals with dementia due to Alzheimer's disease (AD), Parkinson's disease (PD), and Down's syndrome (DS) exhibit similar neuropathological features, including neuritic plaques, neurofibrillary tangles, and the loss of specific populations of neurons. In addition, these brains show similar transmitter-specific neurochemical alterations. Recent evidence indicates that several pathological and neurochemical changes are related to diseases involving projection systems innervating the telencephalon. At least two transmitter-specific circuits, cholinergic neurons in the basal forebrain and noradrenergic neurons in the locus coeruleus, are selectively affected in many patients with these disorders. This review focuses on these systems because they provide particularly useful examples of the ways in which multidisciplinary studies can provide new clinical-pathological-chemical correlations. Strategies used in these studies are applicable to a wide variety of other neurodegenerative diseases affecting specific populations of neurons. Modern neuropathological approaches to these diseases should eventually allow investigators to directly relate pathogenic processes involving transmitter-specific neuronal populations, whose projections, physiological properties, and functions are known, with the clinical manifestations occurring in human disorders of behavior and cognition.

Fluorescence histochemical visualization of catecholamines and immunolabeling of dopamine beta hydroxylase (DBH) were employed to study noradrenergic nerve terminals and perivascular nerve specializations in the rat kidney. Plexuses of catecholamine-containing and dopamine-beta-hydroxylase-immunoreactive nerves innervate the intrarenal arterial tree and larger intrarenal veins. Some perivascular nerve bundles have specialized segments composed of clusters of axonal enlargements that are immunoreactive for DBH and fluoresce intensely in ultraviolet light after fixation in a solution of formaldehyde and glutaraldehyde or treatment with glyoxylic acid. No fluorescent neural structures were found in denervated rat kidney sections treated with glyoxylic acid. Many such structures are associated with arteriolar branches of interlobar, arcuate, and interlobular arteries and are composed, in part, of axonal enlargements that contain mitochondria, microtubules, and one or more clusters of synaptic vesicles. These perivascular nerve specializations may be sites of axoaxonal interactions between noradrenergic axons or between these axons and other types of autonomic or sensory axons. The synaptic vesicles evidently store large amounts of catecholamine, but there is no evidence whether it is released into the surrounding tissue. These structures may be involved in changes in intrarenal innervation patterns which may occur as the rat ages. Regardless of the autonomic or sensory nature of intrarenal neural structures, close association of most such structures with arterioles suggests some neurovascular interaction.