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PubMed1995

Sympathetic and sensory innervation of the rat shoulder joint: a WGA-HRP tracing and CGRP immunohistochemical study.

The innervation of the shoulder joint of the rat was investigated. Nerve origin was assessed by injection of a neuronal tracer (WGA-HRP) into the shoulder joint cavity and calcitonin gene-related peptide (CGRP), which is known to be present in some sensory neurons, was detected immunohistochemically with an anti-CGRP antibody. In the ipsilateral sympathetic and dorsal root ganglia, 133-312 and 12-55 nerve cell bodies were respectively labeled by injection of the tracer. In the sympathetic ganglia, 83% of all labeled cells were found in the stellate ganglion and 17% in the superior cervical ganglion. In the dorsal root ganglia, 75% of the labeled cells were found in C4 and the neighboring ganglia (C4-C5), while the rest were observed in C6-8 and T3. This suggested that the origin of sensory innervation for the shoulder joint was mainly in the mid-cervical cord. CGRP-immunoreactive fibers were found in the synovial capsule of the shoulder joint. These fibers were fine and resembled type 4 axons as classified by Brodal, i.e., nerve related to pain sensation. These findings indicate that sensory nerves from the mid-cervical cord and sympathetic nerves from the cervical ganglion are distributed to the shoulder joint. It is possible that these nerves are related to symptoms such as pain in patients with "frozen" shoulder or other diseases.

Animals↗

PubMed1987

An electron microscope study of quantitative relationships between axon and Schwann cell sheath in myelinated fibres of peripheral nerves.

The quantitative relationships between the cross-sectional area of the Schwann cell sheath (myelin included) and that of its related axon were studied by electron microscopy in the nerve fibres of the spinal roots of lizard (Lacerta muralis). In both ventral and dorsal roots the cross-sectional area of the Schwann cell sheath (myelin included) was found to be directly proportional to that of its related axon (correlation coefficients between 0.88 and 0.92). The ratio between the cross-sectional area of the Schwann cell sheath (myelin included) and that of its related axon tends to diminish as the cross-sectional area of the latter increases. Thus, under normal conditions, in myelinated fibres of the spinal roots of the lizard a quantitative balance exists between the nerve tissue and its associated glial tissue. This result agrees with those previously obtained in the spinal ganglia of the lizard, gecko, cat and rabbit. Some of the mechanisms probably involved in the control of the quantitative balance between nerve tissue and its associated glial tissue in peripheral nerves are presented and discussed.

Animals↗

PubMed1989

Origin of sympathetic and sensory innervation of the knee joint. A retrograde axonal tracing study in the rat.

The origin of sympathetic and sensory innervation of the knee joint was investigated by the intra-axonal transport method. After injection of horseradish peroxidase (HRP) or horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) into the knee joint unilaterally in adult rats, labelled neurons were found ipsilaterally both in sympathetic and dorsal root ganglia at the lower thoracic and lumbosacral levels. A maximum of 1500 and 1000 peroxidase-positive cells were observed in the sympathetic and dorsal root ganglia, respectively; in the former they were mainly located at the levels of L2-L4, and in the latter mainly at L3-L5. The functional implications of the above observations are discussed in relation to pain mechanisms in joints and to the earlier hypothesis that a nervous component might be involved in the pathogenesis of chronic joint inflammation.

Animals↗

PubMed1995

Pax-1, a regulator of sclerotome development is induced by notochord and floor plate signals in avian embryos.

Pax-1 encodes for a DNA-binding transcriptional activator that was originally discovered in murine embryos using a probe from the Drosophila paired-box-containing gene, gooseberry-distal. We have cloned the avian Pax-1 gene as a basis for experimental studies of the induction of Pax-1 in the paraxial mesoderm. The amino acid sequence of the paired-domain is exactly the same in the quail and mouse, whereas outside the paired-domain there is 61% homology. Starting at about the eight-somite stage, quail Pax-1 is expressed in the paraxial mesoderm in a craniocaudal sequence. The unsegmented paraxial mesoderm and the two most recently formed somites do not express Pax-1. In the epithelial somite, the somitocoele cells and the cells of the ventral two-thirds of the epithelial wall are positive. As soon as the sclerotome is formed, only a subset of sclerotome cells expresses Pax-1. These are the cells that migrate towards the notochord to form the perinotochordal tube. Expression then becomes restricted to the intervertebral discs, the perichondrium of the vertebral bodies and the connective tissue surrounding the spinal ganglia. Additional expression domains are found in the scapula and the pelvic region, distinct areas of the head, and the epithelium of the second to the fourth visceral pouch. In later stages the thymus is positive. In vitro and in vivo experiments show that the notochord induces Pax-1 in the paraxial mesoderm, but limb bud mesoderm is not competent to respond to notochordal signals. Floor plate is also capable of inducing Pax-1 expression in sclerotome cells. Our studies show that in competent cells of the paraxial mesoderm, Pax-1 is a mediator of signals emanating from the notochord and the floor plate.

Amino Acid Sequence↗

PubMed1988

Histochemical studies of enzymes of the energy metabolism in postimplantation rat embryos.

Using histochemical procedures for the detection of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), and cytochrome c oxidase (cytox), we investigated the levels of these enzymes of the energy metabolism in postimplantation rat embryos (9.5-12.5 days of gestation). On day 10.5 of gestation, the neural tube, somites, myocardium, and mesenchyme displayed moderate levels of LDH activity; this activity gradually increased in strength, so that, on day 12.5 of gestation, intense LDH activity was uniformly distributed in these intraembryonic tissues. In contrast to LDH, distinct regional differences in the distribution of SDH and cytox were detected. On day 10.5 of gestation, the myocardium exhibited weak to moderate SDH and cytox activity, and on day 11.5, the myocardial activity of these enzymes had become moderate to intense. However, in all other embryonic tissues, e.g., the neural tube and somites, only weak SDH and cytox activity was present. On day 12.5 of gestation, the myocardium displayed very intense SDH and cytox activity, whereas the mantle layer of the neural tube, the spinal ganglia, and the myotomes exhibited only moderate levels of SDH and cytox activity. In the matrix of the neural tube and mesenchyme, these enzyme activities remained at low levels. At electron microscopy, cytox activity was detectable in the spaces between the inner and outer membranes as well as in the intracristal spaces of mitochondria. In general, cytox activity increased in parallel with the differentiation of mitochondria (i.e., increased mitochondrial numbers and size, and the development of mitochondrial cristae), but when the distribution of the cytox activity was considered in detail, it was found to differ among mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1975

Electronmicroscopic study of the effect of 6-an on the sciatic nerve in newborn rats.

12 hrs, 1, 2, and 4 days after i.p. administration of 2, 3 or 5 mg/kg 6-AN (6-Aminonicotinamide) on postnatal day 5, the following findings were made in the sciatic nerve of the newborn rat. In the myelin-forming fibres a swelling between the axolemm and the first inner intraperiod-line of the Schwann cell develops with displacement of the otherwise intact axon. The myelin formation is not impeded. During the 4 days of control after administration of 6-AN there is continual growth of the myelin sheath with an orderly lamellar structure. Using any dose there is no demonstrable alteration in the Schwann cells--with the exception of the inner cytoplasmic tongue--either in the foetal promyelin and mature nonmyelinated fibres. These findings are discussed and compared with the known glia alterations of the spinal ganglia and dorsal radix after the application of 6-AN.

6-Aminonicotinamide↗

PubMed1981

Galactose neuropathy. Permeability studies, mechanism of edema, and mast cell abnormalities.

Galactose neuropathy is characterized by progressive endoneurial edema manifested by a gradual increase in endoneurial fluid pressure. Edema accumulates via a unique mechanism of osmotic force generated by products of the polyol pathway, synthesized within the endoneurial compartment. This paper presents morphologic findings showing firstly, that blood nerve barrier permeability to horseradish peroxidase complexes appears unchanged and secondly, peripheral nerve edema in this condition is restricted to extraganglionic endoneurium sparing the spinal ganglia and adjacent roots. Thirdly, mast cells accumulated in significant numbers and electron microscopy revealed degranulation. There was no evidence of edema in Schwann cell cytoplasm, the putative site of galactitol accumulation via the sorbitol pathway. These findings are discussed with respect to diabetic neuropathy for which galactose intoxication is a useful experimental model.

Animals↗

PubMed1983

Immunoperoxidase study of astrocytic reaction in Junín virus encephalomyelitis of mice.

Intracerebral inoculation of strain XJ Clon 3 of Junín virus into 1-2-day-old mice resulted in the appearance of viral antigen, detectable by means of the peroxidase antiperoxidase (PAP) technique, within the cytoplasm of the neurons of the cerebral cortex, the basal nuclei, cerebellum, pons, medulla, spinal cord, and spinal ganglia; Junín antigen was likewise observed, although to a lesser extent, in the cytoplasm of astrocytes. The viral antigen was found in highest concentration at the cytoplasmic periphery, near the cell membrane, where complete virions are formed by budding. In some cells a "tigroid" distribution of the antigen was observed, suggestive of its production and concentration within the granular endoplasmic reticulum. In spite of the heavy infection of the neural structures, the neurons did not always show major alterations. By immunolabeling of the glial fibrillary acidic protein (GFAP) by the PAP method, a severe glial response could be seen in infected mice, featuring hyperplasia, hypertrophy, and shape distortion of the astrocytes. This easy labeling was not observed in normal mice of the same age, and suggests the accelerated maturation of the astrocytes and the increased GFAP synthesis by direct action of the virus upon such cells. In view of its specificity, use of the PAP technique for GFAP immunolabeling will most likely replace the traditional metal impregnation methods in the study of the astrocytes. Its utilization would be indicated whenever astroglial changes are suspected in any CNS pathologic condition.

Animals↗

PubMed1977

Localization of organ-specific antigens in the nervous system of the rat.

Localization of organ-specific brain antigens in the central nervous system of the rat has been studied by means of indirect immunofluorescence. Rabbit antiserum against homogenate of rat brain, previously absorbed with normal serum and homogenates of rat organs (kidney, liver, spleen), reacted with the water-soluble antigens of rat brain prepared by extraction with phosphate buffer (pH 7.3) and ultracentrifugation at 50 000 X g to give one band in the immunodiffusion test and 2--3 precipitation arcs in immunoelectrophoresis. There was also a positive reaction with peripheral nerve. The antigen was detectable in all regions of the CNS. Cells with distinct cytoplasmic immunofluorescence were most frequently observed in cerebellar white matter, pons, cerebellar pedunculi, longitudinal tracts of the brain stem. Positive immunofluorecence reaction has appeared in the outer plexiform layer and granular layer of the retina, satelite cells of the spinal root ganglia and Schwann cells. A similar reaction was observed in human, mouse and guinea pig brain slices. Both the morphological and immunochemical reactions are indicative of glial localization of this antigen.

Animals↗

PubMed1986

Changes in sorbitol, myo-inositol and lipid inositol in dorsal root and sympathetic ganglia from streptozotocin-diabetic rats.

Sorbitol, fructose, myo-inositol and lipid inositol concentrations were measured in excised dorsal root and sympathetic ganglia from rats with streptozotocin-induced diabetes, both in the acute stage (1 and 2 weeks after the induction of diabetes) and chronically (after 2 months of diabetes). In comparison with age-matched controls, myo-inositol concentrations were decreased by 26-32% after 1 and 2 weeks but had returned to normal levels at 2 months. Lipid inositol concentrations were normal both in the acutely and chronically diabetic animals. Sorbitol was not detectable in ganglia from diabetic or control animals except for a small quantity (0.05 mumol/g wet weight) in dorsal root ganglia at the 2-month stage. Fructose was present in dorsal root ganglia (1.71-3.53 mumol/g wet weight) at all stages and in sympathetic ganglia (2.18 mumol/g wet weight) at the 8-week stage. The differences in these results from those obtained in peripheral nerve trunks are possibly related to the lack of a blood-nerve barrier in sensory and autonomic ganglia.

Animals↗

PubMed1977

Response to stretch of proprioceptors in adult rat muscles de-efferented at birth.

Hind-limb muscles of new-born rats were de-efferented by removing the lumbosacral spinal cord. Spinal ganglia remained intact, together with their peripheral axon. The presence of sensory terminals in limb spindles, induces the full ultrastructural differentiation of muscle spindles, as has been shown previously. In the present paper we have shown by integrating the sensory discharges in the whole nerve from chronically de-efferented muscles that even several months after birth, muscle proprioceptors (probably mostly spindles) still maintain their basic mechanoreceptor properties. Although the limbs were completely immobilized throughout the whole experimental period, spindles from these chronically de-efferented muscles still responded as slowly adapting receptors. The dynamic component was also present in the integrated neurogram response during stretching. It thus appears that basic functional properties of rat muscle proprioceptor persist even when these receptors differentiate and survive without motor innervation and any adequate functional stimuli, i.e. under conditions of permanent disuse.

Animals↗

PubMed1991

P0 gene expression in cultured Schwann cells.

This study examines the expression of the major myelin protein gene P0 in cultured Schwann cells, grown on their own or in association with neurons. Many freshly dissociated Schwann cells from actively myelinating nerves express Po mRNA in high abundance. If neurons are not present, signal intensity falls markedly with time so that by 7 days in culture only a basal expression is evident which is negligible compared to the level in vivo. Dorsal root ganglia from embryo day 16 (E16) rats contain no significant levels of Po mRNA but when grown in full myelinating medium (containing serum and embryo extract) increasing expression is seen from 4 to 5 days onward even though myelination does not occur until after the second week. In this intervening period the intensity of P0 mRNA expression is lower than that found in the actively myelinating cell. Neurons from sympathetic ganglia are also capable of inducing P0 mRNA expression. Schwann cells in dorsal root ganglia explants grown in serum-free defined medium do not assemble a basal lamina and will not wrap or myelinate axons. Nevertheless P0 mRNA, but not protein, is expressed in levels similar to those found in full myelinating medium prior to myelination. Such Schwann cells also exhibit galactocerebroside and the sulphatide recognised by the 04 antibody. It appears that in defined medium or in myelinating medium prior to myelination axonal signals can induce P0 mRNA expression to a certain degree. However, full up-regulation is usually associated with the rapid membrane expansion accompanying myelination. Whether this augmented up-regulation is due to further axonal signalling or events in the Schwann cell is unknown, but the results suggest that P0 expression can be regulated at several stages of synthesis.

Animals↗

PubMed1993

Co-localization of the beta-subtype of protein kinase C and phosphorylation-dependent immunoreactivity of neurofilaments in intact, decentralized and axotomized rat peripheral neurons.

The localizations of protein kinase C-beta-immunoreactivity and phosphorylation-dependent immunoreactivity of neurofilaments were compared in rat dorsal root, hypogastric, and superior cervical ganglia. In all the ganglia studied, protein kinase C-beta and phosphorylation-dependent immunoreactivity of neurofilaments were co-localized in nerve fibres, and no fibres with only protein kinase C-beta-immunoreactivity or phosphorylation-dependent immunoreactivity of neurofilaments were observed. Most intense perikaryal protein kinase c-beta and phosphorylation-dependent neurofilament-staining were seen in large dorsal root ganglion neurons, whereas in the superior cervical ganglion only very faint protein-kinase C-beta and no phosphorylation-dependent staining was seen in the neuronal cell bodies. Both decentralization and axotomy of the superior cervical ganglion induced an accumulation of protein-kinase C-beta-immunoreactivity and phosphorylation-dependent immunoreactivity of neurofilaments in the majority of neuronal perikarya. The accumulation was first observed at 1-2 days postoperation and it persisted up to 6-10 days postoperation. In strongly labelled decentralized neuronal perikarya, precipitation of immunoreactivity was seen near the cell and nuclear membranes, whereas in axotomized neurons, immunoreactivity was often concentrated as a unipolar clump in the cytoplasm. The results show that protein kinase C-beta-immunoreactivity and phosphorylation-dependent immunoreactivity of neurofilaments are colocalized in intact rat peripheral ganglia and that both accumulate transiently in cell bodies of the superior cervical ganglion after decentralization and axotomy.

Animals↗

PubMed1989

[Complications following cell therapy].

So far, the law in the Federal Republic of Germany still allows the injection of fresh-cell preparations from animals as a roborant to increase the vitality of the organism and to strengthen the body's immune defense system. The use of "sicca-cell" preparations was provisionally forbidden in 1987 by the Federal Health Organization (Bundesgesundheitsamt; BGA). Prohibition of fresh-cell injections would have exceeded the authority of this office, although the same serious reservations also applied in the case of this treatment method. Several publications that have appeared since 1955 have reported serious complications of this therapy, some life-threatening and some even lethal. Two further cases are now added: (1) A woman aged 69 had been receiving treatment with cell injections for 9 years. Immediately after an injection of sicca cells she collapsed and was hospitalized; 7 days thereafter she developed an ascending paralysis with increasing inability to swallow or breathe. She died 25 days after the injection as a consequence of central and peripheral respiratory failure. Autopsy revealed the alterations typical for acute Landry-Guillain-Barré-Strohl syndrome. (2) A 76-year-old healthy woman had been receiving treatment with fresh-cell preparations for several years. After an injection of cell suspensions a painful local swelling was observed. The symptoms were interpreted as the consequence of an iatrogenic local hematoma, and repeated punctures were performed to obtain blood. The patient was transferred to a surgical department for further therapy. Two days after the injection she suddenly died with signs of acute cardiovascular failure. Autopsy revealed the signs of a fulminating clostridial infection and also the characteristic signs of Landry-Guillain-Barré syndrome with involvement of the autonomic nervous system. In both cases the development of an inflammatory process in the peripheral nervous system could be interpreted as an immune-mediated allergic disease, related to the repeated injection of heterologous antigenic material containing nervous tissues. This hypothesis would also explain the two other cases already published and would be consistent with the observed perivenous leukoencephalopathy of the central nervous system. The human disease pictures correspond to the well-established animal models of EAEM (experimental allergic encephalomyelitis) and EAN (experimental allergic neuritis). The pathogenesis is discussed; the major role of the central and peripheral nervous system is stressed, with special reference to the risk of acute autonomic failure. The need for specific autopsy techniques for the investigation of the entire nervous system, including spinal cord, roots, spinal ganglia and peripheral nerves with sympathetic chains, is raised.

Adult↗

PubMed1988

Herpes simplex virus type 1 infection in mice with severe combined immunodeficiency (SCID).

Herpes simplex virus type 1 (HSV-1) infection in mutant mice with severe combined immunodeficiency (SCID mice), i.e., mice in which the differentiation of both T and B lymphocytes is severely impaired, was studied. All control (infected and not treated with antibodies or with immune spleen cells) SCID mice were dead by 17 days after intracutaneous injection in the right midflank with 1 x 10(5) PFU of a virulent HSV-1 strain, Hayashida. Immunization with an avirulent strain of HSV-1 (SKa) did not protect them from death or prolong the survival time. Tissue virus titration of infected mice killed at various times after inoculation detected infectious virus in various organs, dorsal root ganglia, spinal cord, brain, kidney and adrenal gland in addition to the inoculation site of the skin in SCID mice, whereas virus could be detected only in the inoculation site and the nervous tissues in euthymic BALB/c mice, and in the adrenal gland from only one out of 17 nude mice. Human gamma globulin containing neutralizing antibody against HSV-1 prolonged the survival time but did not protect SCID mice from death. Transfer of spleen cells from immunized BALB/c mice protected the infected SCID mice from death. Treatment of spleen cells with anti-Thy 1.2 monoclonal antibody and complement abolished the protection.

Animals↗

PubMed1986

Distribution of the adhesion molecules N-CAM and L1 on peripheral neurons and glia in adult rats.

There is considerable evidence that the cell surface glycoproteins N-CAM and L1 are important mediators of cell-cell adhesion in the nervous system, at least during development. Numerous studies have been devoted to the molecular properties of these proteins and their adhesion role in embryonic and early postnatal development. Much less is known about their importance in mature tissues. A rigorous and comprehensive description of the cell distribution of these molecules in the adult nervous system would clearly form a useful baseline for functional and biochemical studies. In the present work we have addressed this issue and studied the distribution of N-CAM and L1 throughout adult, as opposed to developing, rat peripheral nervous tissue. Particular attention was paid to the ganglia of the enteric nervous system, since adhesion mechanisms within these ganglia are likely to be placed under unusual demands. We report, for the first time, the presence of N-CAM and L1 on mature sensory, sympathetic and enteric neurons in adult rats. Thus, immunostaining of cell suspensions or short-term cultures showed N-CAM and L1 surface labelling on sympathetic and both large and small dorsal root sensory neurons. Both antigens were also present on the surface of enteric neurons in cultures prepared from 10-day-old rats and neonatal guinea pigs. Immunostaining of sections of enteric ganglia from adults indicated that both molecules were also expressed by mature enteric neurons. In sections of mature sciatic nerve neither N-CAM nor L1 immunoreactivity were detected at the site where the plasma membrane of myelinated axons meets the ad-axonal plasma membrane of the myelin-forming Schwann cell. Thus, both N-CAM and L1 were detected on all major classes of peripheral neurons, while their levels in the plasma membrane of myelinated axons may be significantly down-regulated. Similarly, both N-CAM and L1 were present on all major classes of non-myelin-forming peripheral glia in adult rats. This includes the enteric glial cells of the myenteric ganglia, non-myelin-forming Schwann cells in the sciatic nerve, sympathetic trunk and fine autonomic nerves in the gut wall, and the satellite glial cells of sympathetic and dorsal root sensory ganglia. In contrast, myelin-forming Schwann cells did not express detectable levels of N-CAM and only very low levels of L1, which was mainly located near the nodes of Ranvier.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1985

Differential vulnerability of 3 rapidly conducting somatosensory pathways in the dog with vitamin B6 neuropathy.

In anesthetized dogs with chronically implanted cortical electrodes somatic sensory-evoked potentials (SEPs) were produced by electrical stimulation at neural, muscular or cutaneous sites of the contralateral hind leg. Stimulation of the tibial nerve at the calcaneus or of the short flexor muscles of the hind paw caused SEPs having characteristics following activation of rapidly conducting afferents from muscle spindles. Stimulation of the glabrous skin of the central pad resulted in SEPs arriving after a more protracted latency evidently related to activation of afferents from Merkel cells, Krause and Pacinian corpuscles known to be located at these sites. Stimulation of the hairy skin from the dorsal surface of the hindpaw produced a further type of SEP presumably resulting from activation of afferents from receptors of tylotrich hair follicles. Vitamin B6-induced neuropathy involves the selective degeneration of the largest neurons in the spinal ganglia and of associated long peripheral and central neurites performing rapid impulse transmission. In the course of vitamin B6 neuropathy the relatively slow impulse transmission following stimulation of the central pad was more severely impaired than the faster one after activation of afferents from muscle spindles or receptors from hair follicles. This allows us to conclude that in the dog afferents from the glabrous skin of the central pad conduct centrally via the dorsal columns, susceptible to vitamin B6 intoxication, while muscle and hair receptor afferents ascend in the dorsal spinocerebellar and spinocervical tract, respectively, which are vitamin B6 resistant.

Animals↗

PubMed1975

Histamine content and synthesis in central and pheripheral nerve structures during stress.

Histamine content and histidine decarboxylase activity in cortex and hypothalamus, together with histamine content in peripheral nervous structures were examined in normal and electrically stressed guinea-pigs. A significant increase in histidine decarboxylase activity in hypothalamus and cortex together with concomitant decrease in histamine content in hypothalamus have been found. Electric shock causes also a decrease in histamine content in spinal cord, spinal ganglia, dorsal roots and sensory nerve. The function of histamine in nervous system is discussed.

Animals↗