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PubMed2004

Nitric oxide synthase expression and cell changes in dorsal root ganglia and spinal dorsal horn of developing and adult Rana esculenta indicate a role of nitric oxide in limb metamorphosis.

Metamorphosis of amphibians requires reconfiguration of sensory and locomotor neural networks. In view of such plastic changes and implications of nitric oxide (NO) in neural developmental shaping, we examined via histochemistry and immunohistochemistry its synthetic enzyme nitric oxide synthase (NOS) in dorsal root ganglia (DRGs) and dorsal horn of the developing and adult frog Rana esculenta. In limb DRGs, NOS positivity was first and selectively detected just before limb bud appearance, increased during metamorphosis, and was then down-regulated. In adulthood, NOS was expressed in some DRG neurons at all segmental levels. Similar features were detected in the dorsal horn neuropil. In limb DRGs, cell counts in Nissl-stained sections revealed a twofold increase of differentiated neurons during metamorphosis and an additional twofold increase in adulthood. Perikaryal sizes in limb DRGs did not vary during metamorphosis but increased and were more heterogeneous in the adult frog, probably reflecting adaptation to body size. NOS and cell changes during metamorphosis were much less marked in DRGs at other levels. Carbocyanine tracing documented selective labeling of NOS-expressing hindlimb DRG neurons from the spinal nerve at the time of initiation of hindlimb movements. The findings show that, in limb DRG neurons, NOS parallels cell differentiation and limb development during metamorphosis. The data also provide evidence of NOS expression in DRG cells innervating the hindlimbs when sensorimotor circuits become functionally mature. This study indicates a key role of NO production in the maturation of sensory functions that subserves in amphibians the transition from swimming to tetrapod locomotion.

Animals↗
PubMed2000

Long-lasting regulation of galanin, opioid, and other peptides in dorsal root ganglia and spinal cord during experimental polyarthritis.

Mechanisms involved in transition from acute to chronic pain are still not well understood and our means to therapeutically influence this transition are limited. Moreover, very little is known about long-lasting consequences of prolonged exposure to painful stimuli with regard to phenotypic changes and pain experience. In this study we have analyzed long term behavioral and neurochemical effects of intradermal tail injection of heat-killed mycobacterium butyricum suspended in complete Freund's adjuvant. Calcitonin gene-related peptide (CGRP) and galanin mRNA levels were investigated in dorsal root ganglia of polyarthritic rats during the acute (21-) and the remission stage (79 days postinjection), and opioid peptide mRNAs and receptors were studied in the spinal cord. Most of the increases in peptide mRNA levels observed during the acute stage of arthritis were still present in the remission stages. Thus, CGRP and galanin mRNAs in DRGs, and opioid peptide mRNAs and opioid receptors in the spinal cord were still strongly up-regulated, when animals do not exhibit spontaneous pain behavior and inflammation. Hot-plate test in the presence of naloxone, performed in the remission stage, indicated that opiates participate in pain threshold regulation after prolonged painful condition. Finally, X-ray examination revealed a complete destruction of joint structure, thus suggesting a parallel lesion of peripheral nerve endings. These results suggest that in the remission stage of chronic joint inflammation several types of mechanisms are activated aiming at counteracting both inflammatory and neuropathic pain. Thus, opioid systems in the dorsal horn as well as galanin in DRG neurons are upregulated, both alternating pain.

Animals↗
PubMed1975

Retrograde transport of horseradish peroxidase in transected axons. II. Relations between rate of transfer from the site of injury to the perikaryon and onset of chromatolysis.

The sciatic nerves of 2--3 month-old mice, rats and rabbits were crushed and at the same time horseradish peroxidase (HRP) was applied around the crushed region at mid-thigh level. At varying time intervals thereafter, HRP was seen in neurons of the corresponding spinal ganglia and levels of the spinal cord. The rate of ascent of HRP appeared to be similar in the different animals and to be independent of the length of the nerve. The accumulation of HRP was more conspicuous in small than large neurons in the ganglia at early stages after the nerve crush. The arrival of HRP in the perikarya preceded the onset of early morphological changes in the neurons (chromatolysis). The rate of ascent of the signal for chromatolysis appeared to be within the same range as that of the retrograde transport of HRP in the crushed axons. The possibility therefore exists that these two phenomena are in some way interrelated.

Animals↗
PubMed1995

Cervical primary afferent input to vestibulospinal neurons projecting to the cervical dorsal horn: an anterograde and retrograde tracing study in the cat.

Vestibulospinal neurons in the caudal half of the medial and descending vestibular nuclei terminate in the cervical spinal cord, not only in the ventral horn and intermediate zone but also in the dorsal horn. The purpose of the present study was to examine whether the areas containing these vestibulospinal neurons are reached by cervical primary afferents. In one group of experiments, wheat germ agglutinin-horseradish peroxidase conjugate and horseradish peroxidase were pressure injected into spinal ganglia C2-C8 and revealed anterogradely labeled fibers and boutons in the caudal part (caudal to the dorsal cochlear nucleus) of the ipsilateral medial and descending vestibular nuclei. This projection was verified in experiments in which wheat germ agglutinin-horseradish peroxidase conjugate was microiontophoretically injected into the caudal half of either the medial or the descending vestibular nuclei and revealed retrogradely labeled cells only in ipsilateral spinal ganglia C2-C7, with a maximum of cells in C3. In another group of experiments, after microiontophoretic injections of Phaseolus vulgaris leucoagglutinin or Biocytin into either the medial or the descending vestibular nuclei, anterogradely labeled fibers and boutons were present in the cervical spinal cord, mainly bilaterally in the dorsal horn (laminae I-VI) but also, to a lesser extent, in the ventral horn and intermediate zone. The existence of a loop that relays cervical primary afferent information to vestibulospinal neurons projecting to the cervical spinal cord, in particular the dorsal horn, may have implications for vestibular control over local information processing in the cervical dorsal horn.

Afferent Pathways↗
PubMed1978

Neural spread of herpes simplex virus types 1 and 2 in mice after corneal or subcutaneous (footpad) inoculation.

Twelve herpes simplex virus (HSV) strains, 6 of each type, were inoculated subcutaneously into the left hind foot and into the cornea of the right eye of 12-day-old Swiss albino mice. The neural spread of virus to trigeminal and spinal ganglia and to brain and spinal cord was studied by demonstration of infective virus, histology and electron microscopy. Type 1 and type 2 infections seemed to spread equally well by intra-axonal transport. Using a protein tracer (horseradish peroxidase) injected into the same site as the virus, transport of the tracer to neurons corresponding to those infected with virus was observed. The extensive destruction of CNS tissue in the transitional region of the trigeminal root where CNS and the peripheral nervous system meet is discussed with reference to the pathogenesis of HSV encephalitis.

Animals↗
PubMed1985

Effect of capsaicin upon fluoride sensitive acid phosphatases in selected ganglia and spinal cord and upon neuronal size and number in dorsal root ganglion.

A quantitative method for the analytical separation of the fluoride sensitive acid phosphatases of rat spinal cord and peripheral nervous tissues into tartrate-sensitive and tartrate-resistant forms (TSAP and TRAP, respectively) is described. Evidence supporting the use of L-(+)-tartrate rather than fluoride as an inhibitor is presented. The method is used for the quantitative description of the consequences of neonatal capsaicin treatment, and the results appear to justify the attribution of the TRAP activity to the capsaicin sensitive neurons. In the first experiment, rats were killed at weekly intervals after neonatal capsaicin treatment. In controls, both TRAP and TSAP activities in dorsal root ganglia (DRG) increased during the second postnatal week and remained constant thereafter. At all ages (1-4 weeks) TRAP activity was reduced 50-60% in capsaicin-treated DRG. Reduction of TSAP activity was much less. In a second experiment, rats were treated neonatally with capsaicin or vehicle (control) and allowed to grow to adulthood (4 months). TRAP activity was found to be decreased 38% in the dorsal half of the lumbar spinal cord (L3-L5), decreased 33% in nodose ganglion, and unchanged in superior cervical ganglion of the capsaicin treated animals. TSAP activity was unchanged in dorsal spinal cord and superior cervical ganglion and decreased 33% in nodose ganglion. The number of neurons in C8 DRG was found to be reduced 28% in capsaicin treated animals. The loss appeared to be among the small neurons. The number of large neurons was actually increased in ganglia from capsaicin-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Phosphatase↗
PubMed1995

Experimental peripheral neuropathy induced in adult rats by repeated intraperitoneal administration of taxol.

Taxol, a natural extract with antineoplastic properties, is known to be neurotoxic in humans. Its neurotoxicity after systemic administration, however, has never been studied in detail at the morphological level in humans and in animals. In this study we administered taxol intraperitoneally to female Wistar rats and we performed an extended neurophysiological and morphological examination of the peripheral nerves, dorsal root ganglia, spinal rootlets, and spinal cord. The results obtained in this experimental model indicate that taxol induces pathological changes mainly in the peripheral nerves, but they are present also in the ventral and dorsal spinal rootlets and spinal dorsal column fibers. The dorsal root ganglia and spinal cord neurons were, on the contrary, unaffected. The most impressive change induced by systemic taxol administration was intraaxonal neurotubule accumulation. Schwann cells showed signs of "activation" but clear demyelination was not observed. We conclude that with the use of this model it is possible to induce a peripheral neuropathy in the Wistar rat which resembles that reported in humans and which can, therefore, be used to better understand the basic mechanism(s) of taxol toxicity and to evaluate protective strategies in an attempt to reduce it.

Animals↗
PubMed1995

On the influence of the perineuronal microenvironment on the outgrowth of perikaryal projections of spinal ganglion neurons.

While the outgrowth of the slender projections from the perikaryon of spinal ganglion neurons is an intrinsic property of these neurons, it is also influenced by the surrounding microenvironment. To obtain evidence concerning whether the outgrowth of these projections is influenced by one or both components of the perineuronal microenvironment (satellite cells plus extracellular matrix) we have taken advantage of a rare arrangement of these neurons. In the spinal ganglia of adult animals nerve cell bodies are occasionally arranged in pairs, the two nerve cell bodies of the pair being separated by a satellite cell sheet lacking a basal lamina, while along the remaining portions of their surfaces they are enveloped by a satellite cell sheath, in turn surrounded by a basal lamina and connective tissue. By studying these paired nerve cells we have been able to compare, in the same nerve cell body, the extent of the perikaryal projections in surface domains associated only with satellite cells and in surface domains associated with both satellite cells and extracellular matrix. In spinal ganglia of the rat and lizard we have found that the overall development of the perikaryal projections does not differ significantly in either of these surface domains. This finding suggests that neuron-satellite cell interactions rather than factors in the extracellular matrix play a role in promoting the outgrowth of perikaryal projections from spinal ganglion neurons.

Animals↗
PubMed1991

Gamma interferon-like immunoreactive material in rat neurons: evidence against a close relationship to gamma interferon.

Gamma interferon is a potent immunoregulatory peptide produced by activated lymphocytes. Recently, a gamma interferon-like immunoreactive molecule has been demonstrated immunohistochemically in subpopulations of rat neurons. We have now further characterized this molecule. Western blot analysis of spinal ganglia homogenates revealed a single 60,000 mol. wt band that was immunoreactive with monoclonal antibody DB1 directed against rat gamma interferon. A polyclonal antiserum and the monoclonal antibodies DB10 and DB12 failed to detect this band although all antibodies were able to label the major 18,000 mol. wt band of recombinant gamma interferon on the same blots. The 60,000 mol. wt band was selectively present in homogenates from primary sensory and sympathetic ganglia but was absent from the central nervous system and other peripheral organs, corresponding to the reported immunocytochemical distribution of gamma interferon-like immunoreactivity. The 60,000 mol. wt protein does not appear to be glycosylated. It could not be solubilized by detergents such as Triton X-100 and it co-purified with cytoskeleton-enriched preparations. At the nucleic acid level, Northern blot analysis using probes specific for rat gamma interferon mRNA failed to detect specific mRNA in rat spinal ganglia, whereas a strong 1.2 kb signal was detected in activated spleen cells. Functionally, gamma interferon-like immunoreactive material is strongly induced in superior cervical ganglion neurons after preganglionic axotomy of the sympathetic chain, but remains constant or slightly decreases in L5 spinal ganglion neurons after sciatic nerve transection. In contrast, major histocompatibility complex antigens are strongly induced on non-neuronal cells in both systems. We conclude that the neuronal gamma interferon-like immunoreactive material is clearly distinct from lymphocyte-derived gamma interferon and might not be involved in the control of major histocompatibility complex expression on glial cells.

Animals↗
PubMed2003

Substance P in the uterine cervix, dorsal root ganglia and spinal cord during pregnancy and the effect of estrogen on SP synthesis.

Prior to parturition the non-pliable uterine cervix undergoes a ripening process ("softens" and dilates) to allow a timely passage of the fetus at term. The exact mechanism(s) triggering and involved in cervical ripening are unknown, though evidence for a role for sensory neurons and their contained neuropeptides is emerging. Moreover, an apparent increase in neuropeptide immunoreactive nerves occurs in the cervix during pregnancy, maternal serum estrogen levels rise at term and uterine cervix-related L6-S1 dorsal root ganglia (DRG) sensory neurons express estrogen receptor (ER) and neuropeptides. Thus, we sought to test the hypothesis that the neuropeptide substance P (SP) changes biosynthesis and release over pregnancy, that estrogen, acting via the ER pathway, increases synthesis of SP in DRG, and that SP is utilized in cervical ripening at late pregnancy. Using immunohistochemistry, in situ hybridization, reverse transcriptase-polymerase chain reaction (RT-PCR) and radioimmunoassay (RIA), we investigated coexpression of ER-alpha/beta and SP; differential expression of ER-alpha and -beta mRNA in DRG neurons; SP synthesis in DRG; and changes in SP concentration in the cervix, DRG and spinal cord over pregnancy. In addition, the effect of exogenous estrogen on SP synthesis in L6-S1 DRG of ovariectomized rats was examined. SP-immunoreactive neurons expressed ER-alpha and ER-beta. SP synthesis (expressed as beta-PPT mRNA label) was prominent in small DRG neurons. SP concentration increased in the L6-S1 DRG and spinal cord segments, with a peak at Day 20 of gestation, but decreased in the cervix during the first two trimesters, with a rise over the last trimester to Day 10 levels. SP and ER-alpha mRNA synthesis increased in DRG over pregnancy but ER-beta mRNA levels were largely unchanged. When ovariectomized rats were treated with exogenous estrogen, SP mRNA synthesis in the DRG increased in a dose-related manner, an effect blocked by ER blocker ICI 182 780. From these results, we postulate that synthesis of SP in L6-S1 DRG and utilization in the cervix increase over pregnancy and this synthesis is under influence of the estrogen-ER system, most likely ER-alpha. We postulate that SP may play a role in cervical ripening and, consequently in the birth process.

Animals↗
PubMed1985

Free amino acids in the spinal cord and ganglia of dogs after ligation of the abdominal aorta.

The authors studied the effect of 40 min and 6 days occlusion of the abdominal aorta on the aspartate [Asp], glutamic acid [Glu], glutamine [Gln], glycine [Gly] and alanine [Ala] concentration in both parts of the grey matter of the lumbosacral cord, in the spinal ganglia and along the dorsal fasciculi. After 40 min ischaemia, an increase was found in the Glu, Gln, Gly and Ala concentration in the dorsal part of the grey matter and in the Glu, Gly and Ala concentration in the spinal ganglia. In the ventral part of the grey matter only the Ala concentration was increased. After 6-day ligation, the Asp and Gly concentration fell in the ventral horns, while the Gly concentration rose in the dorsal horns. The Ala and Asp concentration in the spinal ganglia rose. After this interval the Asp and Glu concentration also rose in the fasciculus gracilis. The Ala/Glu concentration ratio showed the most pronounced increase in the central horns of the grey matter.

Alanine↗