Components Of Peripheral Nervous System Research Articles

Normal segmentation and size of the primary sympathetic ganglia depend upon the alternation of rostrocaudal properties of the somites

Metameric organization of the dorsal root ganglia (DRG) and ventral roots depends on the alternation of rostrocaudal properties within the somites. In addition, the size of DRG is likely to be regulated by the adjacent mesoderm, because unilateral creation of a paraxial mesoderm with only rostral somitic (RS) halves, leads to the development of non-segmented DRG that are larger and contain more cells than the sum of the contralateral, control DRG. We have now extended our studies of the role of the paraxial mesoderm in the morphogenesis of the peripheral nervous system (PNS) to another metameric PNS component, the sympathetic ganglia (SG). The development of the primary sympathetic chain was studied in chick-quail chimeras with multiple half-somite grafts using quantitative morphometric analysis. In the presence of an exclusively rostral or caudal somitic mesoderm, segmentation of the initially homogeneous primary sympathetic chain into ganglia is prevented. Therefore, the SG, like the DRG and ventral roots, require the normal rostrocaudal alternation of the somitic mesoderm for segmental morphogenesis. On embryonic day 4 (E4), there is a 38% average decrease in the volume of the primary sympathetic chain opposite a RS mesoderm, compared with the primary chain on the unoperated side. This is in contrast to the average increase of 27% in the volume of the DRG opposite the grafted mesoderm in the same embryos. Our results, and classical observations, have led us to propose a model in which the mesoderm controls DRG and SG size by modulating the partition of migrating NC precursors between the anlage of these two ganglion types. According to this model, the reduction in SG volume and concomitant increase in DRG volume observed opposite RS grafts, results from the arrest in the DRG anlage of neural crest cells that normally migrate to the SG.

THE PATHOPHYSIOLOGY OF CHRONIC RELAPSING EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS IN THE LEWIS RAT

Electrophysiological studies were performed in Lewis rats with chronic relapsing experimental allergic encephalomyelitis (EAE) induced by inoculation with guinea-pig spinal cord and adjuvants and treatment with low dose cyclosporin A. During clinical episodes there was conduction failure in the central nervous system (CNS), namely the spinal cord dorsal columns, and in the afferent fibres in the peripheral nervous system (PNS). The following observations indicated that the conduction failure was mainly due to demyelination-induced conduction block: (1) rate-dependent conduction block in the CNS and PNS; (2) temporal dispersion due to slowing of PNS conduction; (3) restoration of PNS conduction by cooling; (4) restoration of CNS conduction by ouabain; (5) previously demonstrated histological evidence of primary demyelination in the dorsal columns, dorsal root ganglia and dorsal roots; and (6) the temporal association of restoration of conduction with remyelination. However, it is likely that CNS and PNS axonal degeneration, which occurs in this disease, also contributed to the conduction failure. In clinical remissions there was restoration of conduction in the CNS and PNS which can be explained by ensheathment/remyelination by oligodendrocytes and Schwann cells, respectively. In most rats during clinical episodes the cerebral somatosensory evoked potential was reduced in amplitude and prolonged in latency, which can be accounted for by demyelination and axonal degeneration in the CNS and PNS components of the afferent pathway. In 2 rats with episodes of EAE, however, this potential was markedly increased in amplitude, which might have been due to demyelination-induced conduction block of descending pathways that normally inhibit synaptic transmission in the afferent pathway. In well-established remission there was residual conduction failure in the CNS and PNS which can be mainly accounted for by axonal degeneration.

Extracellular matrix components of the peripheral pathway of chick trigeminal axons.

The directed growth of axons to their peripheral targets during development may be influenced by a variety of intrinsic and environmental factors, the latter including the pattern of extracellular matrix components in the region through which they grow. We investigated the localization within the chick trigeminal mesenchyme of a variety of extracellular matrix molecules (laminin, heparan sulfate proteoglycan, entactin, collagen type IV) by using indirect immunofluorescence techniques. The trigeminal mesenchyme contained several of these molecules, but only laminin was specifically localized to trigeminal axon pathways. Double immunofluorescence localization of affinity-purified laminin antibodies and monoclonal antibodies directed against a neuron-specific beta-tubulin (to visualize growing axons and postmitotic V ganglion neurons) demonstrated that the V motor and sensory peripheral nerves confine their growth to two patches of laminin-rich mesenchyme--a ganglionic laminin patch colocalized with V ganglion neurons and their axons, and a mandibular laminin patch colocalized with the V motor axons. Thus, laminin pathways in the mesenchyme may help guide V axons along their appropriate routes in the periphery. Double immunofluorescence localization of these laminin antibodies and monoclonal antibodies that recognize neural crest cells (to visualize precursors of V ganglion neurons and glia) demonstrated that the majority of cells within the ganglionic laminin patch were neural crest and differentiating neurons derived from the trigeminal epidermal placode. Since these cells often were laminin-immunopositive, they might be a source of the ganglionic laminin matrix. The mandibular laminin patch contains nearly no neural crest cells, and probably contains the somitomeric precursors to the mandibular muscle mass. These results suggest that laminin, a matrix molecule implicated in the guidance of trigeminal peripheral axons, might be produced in localized patches by peripheral nervous system components and by muscle targets.

Identification of a cell surface glycoprotein family of olfactory receptor neurons with a monoclonal antibody

A monoclonal antibody (Mab) has been developed which recognizes a family of cell surface glycoproteins found in high levels of rat olfactory receptor neurons. This Mab, designated 2B8, was produced by the fusion of X63-Ag8.653 myeloma cells and spleen cells of a mouse immunized with PC12 rat pheochromocytoma cells. Immunofluorescence analyses of cryostat sections of neonatal olfactory epithelium show prominent 2B8 binding to receptor neurons. Within the olfactory bulb only the glomerular and olfactory nerve layers show 2B8 binding. All other neural structures in the main olfactory bulb have background levels of reactivity. Analyses of 2B8 binding to particulate protein preparations from several central and peripheral nervous system components demonstrated highest 2B8 antigen specific activity in olfactory bulb and epithelium and detectable levels in dorsal root ganglia (DRG), whole cerebrum, cerebellum, and brainstem. However, 2B8 antigen could not be detected in non-olfactory structures by immunofluorescence. Some non-neural tissues also had the ability to bind 2B8 Mab in the particulate protein radioimmunoassay. In order to compare the 2B8-reactive molecules found in each tissue, Mab was applied to polyacrylamide gels of unlabeled membrane proteins. A family of molecules with diverse molecular weights was found. Some were unique to individual tissues whereas others were shared among tissues. Olfactory bulb and epithelium had a unique band with Mr = 215,000 and another band with Mr = 142,000. The 142,000-dalton band was also found with PC12 cells. PC12 cells also had several bands of lesser molecular weight, including 51,000 and 43,000. Testes membranes had immunoreactive bands only at Mr = 46,000 and 43,000. Bone marrow, perinatal liver, and DRG each expressed a single 2B8-reactive band with Mr = approximately 114,000. Salivary gland had four reactive bands, two common to it and only PC12 cells, the 114,000-dalton band which is similar to that found in adult rat bone marrow and DRG, and a unique band at Mr = 152,000. 2B8 immunoprecipitates of olfactory bulb and epithelium were analyzed for glycosyl groups by lectin reactivity. Wheat germ agglutinin and Ricinus communus agglutinin I bound the 2B8 antigens using two distinct assay methods. This suggests that the 2B8 antigens recognized in the olfactory system are glycoproteins having sialic acid and D-galactosyl components.(ABSTRACT TRUNCATED AT 400 WORDS)

PERIPHERAL NERVE CONDUCTION STUDIES IN PASSIVELY ADDICTED NEONATES

The contribution of the peripheral nervous system to the neonatal abstinence syndrome was evaluated. Nerve conduction velocities were determined in 25 passively addicted(PA)infants born to mothers receiving methadone maintenance; 12 of the mothers abused other drugs concomitantly. Nerve conduction velocities in PA infants, both at 3-7 days and 3-4 weeks of age, did not differ from those of 11 control infants. Within the PA group, no conduction velocity differences(p > 0.1) were noted when comparing (a)maternal methadone use versus polydrug abuse, (b) mild versus severe neonatal abstinence, and (c)neonatal treatment with either paregoric or phenobarbital. Nerve conduction velocities were also normal in 3 PA infants who developed abstinence-related seizures. No relationship could be demonstrated between nerve conduction velocities and severity of abstinence on day of study(p> 0.4). Electromyographic studies were normal in 23 PA infants; the remaining two infants displayed minimal denervation abnormalities. Since 2-5 year follow-ups in PA infants have shown motor dysfunction and since this study fails to document a peripheral nervous system component to this dysfunction, the role of the central nervous system should be the prime focus of future research in the neonatal abstinence syndrome.

Spasmodic Torticollis as a Cause of Ulnar Nerve Entrapment

Abstract Spasmodic torticollis is thought to be a central nervous system disorder with no peripheral nervous system component. Several consecutive patients with spastic torticollis were found to have ulnar nerve compromise at the elbow on the side ipsilateral to the direction of chin turning. This article describes three such patients in detail. It is postulated that the cause is the stereotypical posture assumed by these patients in using their hand to maintain a neutral head position. The chronic leaning and elbow flexion place the ulnar nerve at risk for the development of pressure neuropathy.

Spasmodic torticollis as a cause of ulnar nerve entrapment.

Spasmodic torticollis is thought to be a central nervous system disorder with no peripheral nervous system component. Several consecutive patients with spastic torticollis were found to have ulnar nerve compromise at the elbow on the side ipsilateral to the direction of chin turning. This article describes three such patients in detail. It is postulated that the cause is the stereotypical posture assumed by these patients in using their hand to maintain a neutral head position. The chronic leaning and elbow flexion place the ulnar nerve at risk for the development of pressure neuropathy.

Induction of expression of the rat G5 nervous system antigen occurs postnatally

G5 is a cell membrane component found in high levels in the adult rat central nervous system and in low levels on some cells of the rat immune system. Binding assays with adult brain particulate protein preparations and monoclonal antibody to G5 (G5-IgG) gave highest activity in cerebral cortex and lowest in the white matter rich regions pons and spinal cord. Three peripheral nervous system components had no G5 activity. Analysis of G5 content in particulate protein preparations from whole rat brain of ages embryonic day 15 to adult indicates that G5 is present in negligible amounts in the newborn rat. It increases in both specific and total activity to reach adult levels by postnatal day 30. A similar induction curve was observed with cerebellum samples. In adult cerebellum, high levels of G5 were found in the molecular layer using both autoradiographic and immunofluorescence techniques. White matter had essentially no activity. The density and uniformity of reaction product in these techniques suggest that G5 is present on a major cellular constituent of the molecular layer. Pharmacologic experiments indicate G5 is not detectable on climbing fibers on adrenergic fibers. Cerebellar samples from juvenile rats also had predominant G5 activity in the forming molecular layer.

Early segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium

This article addresses the problem of the segregation of cell lines during the development of peripheral nervous system components from the neural crest. We show here that committed precursors of peripheral neurons are present in the crest before the migration of its cells has started. If cultured in a serum-deprived medium, a subpopulation of the crest cells readily differentiates into neurons without dividing. Neuronal markers such as neurofilament proteins and receptor sites for tetanus toxin are not expressed in the committed neuronal precursors, but appear after a few hours in culture. They are coexpressed in neurons with the mesenchymal intermediate filament protein, vimentin, which is common to all neural crest cells regardless of their prospective fate. A strong inhibitory effect of serum factor(s) on neurite outgrowth is demonstrated. We show also that conditions stimulating proliferation of crest cells are incompatible with promotion of neuronal differentiation and viceversa.

Biochemical demonstration of the myelin-associated glycoprotein in the peripheral nervous system.

Recent immunocytochemical studies indicated that the myelin-associated glycoprotein (MAG) is localized in the periaxonal region of central nervous system (CNS) and peripheral nervous system (PNS) myelin sheaths but previous biochemical studies had not demonstrated the presence of MAG in peripheral nerve. The glycoproteins in rat sciatic nerves were heavily labeled by injection of [3H]fucose in order to re-examine whether MAG could be detected chemically in peripheral nerve. Myelin and a myelin-related fraction, W1, were isolated from the nerves. Labeled glycoproteins in the PNS fractions were extracted by the lithium diiodosalicylate (LIS)-phenol procedure, and the extracts were treated with antiserum prepared to CNS MAG in a double antibody precipitation. This resulted in the immune precipitation of a single [3H]fucose-labeled glycoprotein with electrophoretic mobility very similar to that of [14C]fucose-labeled MAG from rat brain. A sensitive peptide mapping procedure involving iodination with Bolton-Hunter reagent and autoradiography was used to compare the peptide maps generated by limited proteolysis from this PNS component and CNS MAG. The peptide maps produced by three distinct proteases were virtually identical for the two glycoproteins, showing that the PNS glycoprotein is MAG. The MAG in the PNS myelin and W1 fractions was also demonstrated by Coomassie blue and periodic acid-Schiff staining of gels on which the whole LIS-phenol extracts were electrophoresed, and densitometric scanning of the gels indicated that both fractions contained substantially less MAG than purified rat brain myelin. The presence of MAG in the periaxonal region of both peripheral and central myelin sheaths is consistent with a similar involvement of this glycoprotein in axon-sheath cell interactions in the PNS and CNS.

Autoradiographic and ultrastructural studies of areas of spinal cord occupied by Schwann cells and Schwann cell myelin

Schwann cells, peripheral-type myelin and connective tissue elements develop within the dorsal portion of the X-irradiated spinal cord in immature rats. Factors controlling the distribution of these elements within the irradiated site are not fully understood. In the present study [ 3H]thymidine autoradiography was used to examine proliferative activities of cells in these areas occupied by peripheral nervous system components, and correlative ultrastructural evaluations were made. At 15 and 20 days post-irradiation (P-I), the Schwann cells occupied the dorsolateral portions of the dorsal funiculi, and heavily labeled cells occurred throughout these areas. By 25 days P-I the Schwann cells extended ventrally into the depths of the dorsal funiculi and into the dorsal gray matter, and labeled cells were concentrated in the deeper portions of these areas. Ultrastructurally, the Schwann cells and peripheral-type myelin were more mature in the superficial portions where proliferative activity was diminished. In contrast, much less mature, peripheral-type myelin occurred in the depths where the labeled cells were concentrated. At 30 and 45 days P-I, labeled cells were much less frequent but usually occurred in the depths when observed. Similarly, a dorsal-ventral gradient in maturity of peripheral-type myelin was evident ultrastructurally. By 60 and 90 days P-I, labeling was rare, and mature Schwann cell myelin was present throughout the areas. Astrocytes and their processes were less numerous in regions invaded by Schwann cells, as compared to controls, and studies are in progress to evaluate the relationships between these glial elements and intraspinal peripheral nervous system components.