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Kjell Fuxe - One of the best experts on this subject based on the ideXlab platform.
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adrenalectomy increases the number of substance p and somatostatin immunoreactive nerve cells in the rat Lumbar dorsal root Ganglia
Brain Research, 1994Co-Authors: Rafael Covenas, M Deleon, Gerson Chadi, A Cintra, Janake Gustafsson, J A Narvaez, Kjell FuxeAbstract:Using an immunocytochemical technique we have analyzed changes in substance P, somatostatin, calcitonin gene-related peptide, and galanin immunoreactivity pattern in the rat dorsal root Ganglia. After 7 days of adrenalectomy, sham operated rats were compared with adrenalectomized animals either receiving a daily intraperitoneal injection of 10 mg/kg b.wt. corticosteronev or vehicle. Three Lumbar Ganglia from each animal were blocked, serially cut, and immunostained for each neuropeptide by means of the biotin-avidin-peroxidase technique. A systematic sampling of immunoreactive ganglion cells was performed and the sample number of immunoreactive ganglion cells was calculated. After adrenalectomy, the number of substance P and somatostatin immunoreactive ganglion cells markedly increased ((means±S.E.M.): 245 ± 68versus123 ± 12 for sham operated animals, P < 0.01 (substance P) and 42 ± 8 as compared to 22 ± 9 for sham operated animals, P < 0.01 (somatostatin)). No significant changes were found in the number of calcitonin gene-related peptide and galanin immunoreactive cells after adrenalectomu. These results suggest that adrenal steroid hormones may reduce the synthesis of both substance P and somatostatin in the dorsal root ganglion cells. Daily treatment with a high dose of corticosterone, mimicking its serum levels after stress, failed to prevent the increase of peptide contents after adrenalectomy. These observations also indicate that a tonic action of corticosterone on mineralocorticoid receptors may be crucial for peptide regulation in the spinal Ganglia. These results may be of relevance to adrenalectomy induced changes in sensory mechanisms, neurogenic inflammation and pain transmission and to a role of substance P and somatostatin in these processes.
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short communication adrenalectomy increases the number of substance p and somatostatin immunoreactive nerve cells in the rat Lumbar dorsal root Ganglia
1994Co-Authors: Gerson Chadi, A Cintra, Kjell FuxeAbstract:Using an immunocytochemical technique we have analyzed changes in substance P, somatostatin, caicitonin gene-related peptide, and galanin immunoreactivity pattern in the rat dorsal root Ganglia. After 7 days of adrenalectomy, sham operated rats were compared with adrenalectomized animals either receiving a daily intraperitoneal injection of 10 mg/kg b.wt. corticosterone or vehicle. Three Lumbar Ganglia from each animal were blocked, serially cut, and immunostained for each neuropeptide by means of the biotin-avidin-peroxidase technique. A systematic sampling of immunoreactive ganglion cells was performed and the sample number of immunoreactive ganglion cells was calculated. After adrenalectomy, the number of substance P and Somatostatin immunoreactive ganglion cells markedly increased ((means + S.E.M.): 245 ___ 68 versus 123 + 12 for sham operated animals, P < 0.01 (substance P) and 42 + 8 as compared to 22 + 9 for sham operated animals, P < 0.01 (somatostatin)). No significant changes were found in the number of calcitonin gene-related peptide and galanin immunoreactive cells after adrenalectomy. These results suggest that adrenal steroid hormones may reduce the synthesis of both substance P and somatostatin in the dorsal root ganglion cells. Daily treatment with a high dose of corticosterone, mimicking its serum levels after stress, failed to prevent the increase of peptide contents after adrenalectomy. These observations also indicate that a tonic action of corticosterone on mineralocorticoid receptors may be crucial for peptide regulation in the spinal Ganglia. These results may be of relevance to adrenalectomy induced changes in sensory mechanisms, neurogenic inflammation and pain transmission and to a role of substance P and somatostatin in these processes.
Jozef Maršala - One of the best experts on this subject based on the ideXlab platform.
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Immunohistochemical, Histochemical and Radioassay Analysis of Nitric Oxide Synthase Immunoreactivity in the Lumbar and Sacral Dorsal Root Ganglia of the Dog
Cellular and Molecular Neurobiology, 2006Co-Authors: Nadežda Lukacova, Dalibor Kolesar, Martin Marsala, Jozef MaršalaAbstract:In this study, immunohistochemistry for neuronal nitric oxide synthase (bNOS-IR), nicotinamide adenine dinucleotide phosphate diaphorase histochemistry (NADPHd) and nitric oxide synthase radioassay were used to study the occurrence, number and distribution pattern of nitric oxide synthesizing neurons in the Lumbar (L1–L7) and sacral (S1–S3) dorsal root Ganglia of the dog. Nitric oxide synthase immunolabelling was present in a large number of small- (area 2000 μm^2) neurons. Although neuronal nitric oxide synthase immunolabelling and histochemical staining provided intense staining of multiple small- and medium-sized neurons in all Lumbar and sacral dorsal root Ganglia, immunolabelled or histochemically stained somata exhibited little topographic distribution in individual dorsal root Ganglia. Great heterogeneity was noticed in the immunolabelling of medium-sized nitric oxide synthase immunopositive neurons ranging from lightly immunolabelled somata to heavily immunoreactive ones with completely obscured nuclei. Both staining procedures proved to be highly effective in visualizing intraganglionic fibers of various diameters. In general, the largest fibers revealed at the peripheral end of Lumbar and sacral dorsal root Ganglia were larger, 6.49–9.35 μm in diameter, while those running centrally and proceeding into the dorsal roots were about 30% reduced, ranging between 5.32 and 8.67 μm in diameter. Peripherally, the occurrence of nitric oxide synthase detected in axonal profiles, and confirmed histochemically, in the specimens of the femoral and sciatic nerves, is the first indication of the presence of nitric oxide synthase in the peripheral processes of somata located in L4–S2 dorsal root Ganglia. Large and thin central nitric oxide synthase immunoreactive processes of L1–S3 dorsal root ganglion neurons segregate shortly before entering the spinal cord, the former making a massive medial bundle in the dorsal root accompanied by a slim lateral bundle penetrating Lissauer's tract. Quantitative assessment of the distribution of bNOS-IR and/or NADPHd-stained neurons showed a peculiar pattern in relation to spinal levels. Apparent incongruity was found in the total number of NADPHd-stained versus bNOS-IR neurons, demonstrating a clear prevalence of small bNOS-IR somata in all Lumbar Ganglia, while medium-sized NADPHd-stained somata clearly prevailed all along the rostrocaudal axis with a peak in L5 ganglion. While the number of small bNOS-IR neurons clearly outnumbered NADPHd-stained and NADPHd-unstained somata in S1–S3 Ganglia, an inverse relation appeared comparing the total number of medium-sized NADPHd-stained and NADPHd-unstained somata compared with the number of moderate and intense bNOS-IR neurons. Densitometry of bNOS-IR and NADPHd-stained neurons in Lumbar and sacral Ganglia revealed two distinct subsets of densitometric profiles, one relating to more often found medium-sized bNOS immunolabelled and the other, characteristic for moderately bNOS immunoreactive somata of the same cell size. Considerable differences in catalytic nitric oxide synthase activity, determined by conversion of [^3H]arginine to [^3H]citrulline were obtained in lumbosacral dorsal root Ganglia all along the lumbosacral intumescence, the lowest (0.898± 0.2 dpm/min/μg protein) being in the L4 dorsal root ganglion and the highest (4.194± 0.2 dpm/min/μg protein) in the S2 dorsal root ganglion.
Junming Zhang - One of the best experts on this subject based on the ideXlab platform.
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increased sensitivity of sensory neurons to tumor necrosis factor α in rats with chronic compression of the Lumbar Ganglia
Journal of Neurophysiology, 2002Co-Authors: Baogang Liu, S J Brull, Junming ZhangAbstract:Proinflammatory cytokines may sensitize primary sensory neurons and facilitate development of neuropathic pain processes after peripheral nerve injury. The goal of this study was to determine wheth...
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a comparison of chronic pain behavior following local application of tumor necrosis factor α to the normal and mechanically compressed Lumbar Ganglia in the rat
Pain, 2002Co-Authors: Yuko Homma, S J Brull, Junming ZhangAbstract:To study the role of inflammatory cytokines in the initiation and persistence of radiculopathy as seen in humans, tumor necrosis factor alpha (TNF-alpha) was administered either to normal, uninjured L5 dorsal root Ganglia (DRG) of rats via a hole drilled through the transverse process, or to chronically compressed L5 DRG via a hollow stainless steel rod inserted into the intervertebral foramen. In other experiments, a mixture of soluble TNF receptors (sTNF-Rs: sTNF-RIplus minussTNF-RII) was locally delivered to the chronically or acutely compressed DRG to neutralize the activity of endogenous TNF-alpha. Behavioral tests of mechanical allodynia were performed before and after TNF-alpha administration. Infusion of the normal DRG with TNF-alpha at a rate of 1 microl/h for 7 days induced ipsilateral mechanical allodynia (i.e. decreased mechanical withdrawal threshold) that lasted about 2 weeks. Infusion of the compressed DRG did not alter compression-induced allodynia within the first operative week but substantially enhanced the ipsilateral allodynia after the first postoperative week. Neutralizing the activity of endogenous TNF-alpha of the compressed DRG with sTNF-Rs reduced allodynia for 3 days, but was subsequently without effect. Similar results were obtained when sTNF-Rs were chronically administrated at the acutely compressed ganglion. Results demonstrated that exogenous TNF-alpha causes pain and mechanical allodynia when deposited at the normal DRG, and further enhances the ongoing allodynia when administrated at the compressed DRG. Results also suggest that endogenous TNF-alpha contributes to the early development of mechanical allodynia in rats with chronic DRG compression.
Aldo Rustioni - One of the best experts on this subject based on the ideXlab platform.
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dorsal root ganglion neurons projecting to the dorsal column nuclei of rats
The Journal of Comparative Neurology, 1992Co-Authors: Rosario Giuffrida, Aldo RustioniAbstract:Dorsal root ganglion (DRG) neurons may give origin to ascending branches that terminate in the dorsal column nuclei (DCN); uncertainties still exist with regard to the proportion of these neurons in different DRGs and to the type of these neurons. The percentage and size of neurons that project to the DCN were determined in a large number of DRGs by means of the retrograde transport of colloidal gold-labeled wheat germ agglutinin conjugated to enzymatically inactive horseradish peroxidase (WGAapoHRP-AU). A total of 16,239 neurons was tallied in 80 DRGs from nine rats; 3,240 (20%) of these were retrogradely labeled by the tracer injected in the DCN. Percentages of DCN projecting neurons vary considerably at different segmental levels: they are higher in cervical (up to 63%) than in thoracic (up to 31% for T1, up to 12% for thoracic DRGs below T1) or Lumbar DRGs (up to 15%). At cervical levels highest percentages were encountered in C6, C7, and C8 and lowest percentages in C2-C4. At Lumbar levels highest percentages were encountered in L4 and lowest in L1 and L6. When considering the soma size of DRG neurons it appears that: (1) there are more large cells, labeled and unlabeled, at cervical (38%) than at Lumbar levels (30%) and more at Lumbar than at thoracic levels (23%); (2) at every level, most labeled, i.e., projecting, neurons are large; and (3) DRGs with the highest proportions of large vs. small cells contain the highest percentages of DCN projecting neurons. These results represent the first attempt at establishing the percentages and soma size of DCN projecting neurons from a large number of DRGs and at comparing the contribution to these nuclei from cervical, thoracic, and Lumbar DRGs. Some of the differences in the ratio of projecting neurons at different levels may be explained on the basis of well-known anatomical features, e.g., the projections to the Clarke's column of many DRG neurons in Lumbar Ganglia. The contribution of virtually exclusively large DRG neurons to the DCN, suggested by indirect or incomplete evidence, is demonstrated by the present retrograde labeling and soma size measurements. The results relate to the functional component of peripheral receptors that relay their input via the dorsal columns and do not seem to support a recent suggestion that a sizeable fraction of unmyelinated primary afferents ascend in the dorsal columns to terminate in the DCN.
Vaclav Brazda - One of the best experts on this subject based on the ideXlab platform.
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dynamics of interleukin 6 elevation in rat Lumbar Ganglia of rheumatoid arthritis model
2014Co-Authors: B Madrova, Ilona Klusakova, Vaclav Brazda, Petr DubovýAbstract:Rheumatoid arthritis (RA) is autoimmune inflammatory disease that affected many tissues and organs. RA is associated with an increased production of a range of cytokines. Interleukin 6 (IL-6) is an injury-induced factor that contributes significantly to maintenance of neuropathic pain. The aim of present study was to determine the temporal changes of IL-6 protein in L3 and L5 dorsal root Ganglia (DRG) related with afferent innervation of the knee joint in AIA model. In addition, we monitored manifestation of neuropathic pain using behavioral test. For induction of unilateral antigen-induced arthritis (AIA), rats (n=33) were immunized by an intra-articular injection of Complete Freund’s Adjuvant (CFA, 50 µl) into the right knee joint. The knee joints of naive and control rats (n=15) injected with 50 µl of sterile saline were used for comparison with AIA induction. All experimental animals were left to survive for 1, 2, 4, 14 and 21 days. Upregulation of IL-6 protein in DRG was analyzed by immunofluorescence staining and quantified by Western blot analysis. Neuropathic pain expressed by mechanical hyperalgesia was measured using the dynamic plantar esthesiometer. For thermal hyperalgesia, withdrawal time was measured and the intensity radiance was set to a value of 50. The data differences were evaluated by Mann-Whitney test using STATISTICA software. Increasing of IL-6 protein was found in both L3 DRG and L5 DRG compared with DRG from naive rats. Elevation of IL-6 protein level was observed in both ipsilateral and contralateral L3 DRG in the overall time period except in contralateral L3 DRG at the first day. L5 DRG also showed bilateral but delayed increase in IL-6 protein compare to L3 DRG. Statistically insignificant IL-6 elevation was in ipsilateral L5 DRG during the first two days and in contralateral L5 DRG in the first day. Mechanical allodynia was measured in the ipsilateral hind paws from 1 day to 14 days, in the contralateral hind paws in the period from 7 to 14 days after RA induction. By contrast, hypersensitivity for thermal stimuli was monitored bilaterally during the first fourth days and then 21 day, in the period between the 4 and 14 days has occurred normalization to value before CFA application. The difference at the contralateral side against the ipsilateral hind paws is that a statistically significant decrease was observed only at 4 and 21 days, after the application CFA. Dynamic of IL-6 elevation was found bilaterally in L3 and L5 DRGs of rats subjected by unilateral AIA in correlation with hyperalgesia from 1 to 21 days. The results of increasing of IL-6 protein in L3 and L5 DRGs are related with afferent innervation of ventral and dorsal part of the knee joint. Supported by the project CEITEC (CZ.1.05/1.1.00/02.0068) from European Regional Development Fund.
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ab0113 dynamics of macrophage activation in rat Lumbar Ganglia of rheumatoid arthritis model
Annals of the Rheumatic Diseases, 2014Co-Authors: B Madrova, Petr Dubový, Ilona Klusakova, Vaclav BrazdaAbstract:Background Rheumatoid arthritis (RA) is a chronic, autoimmune inflammatory disease that affects approximately 1% of the population. The early stages of RA are characterized by activation of innate immunity including activation of macrophages (1). Activated macrophages, either resident or recruited from the blood vessels, may release widespread inflammatory mediators that can critically contribute to acute and chronic disease (2). Objectives The aim of our present study was to compare a distribution of activated macrophages in the Lumbar dorsal root Ganglia (DRGs) and their number changes in time of survival. In addition, we monitored manifestation of neuropathic pain using behavioral test. Methods For induction of unilateral antigen-induced arthritis (AIA), rats (n=42) were immunized by an intra-articular injection of Complete Freund9s Adjuvant (CFA, 50 μl) into the right knee joint. The knee joints of control rats were injected with 50 μl of sterile saline. Naive rats were without AIA induction and injection of saline. All experimental animals were left to survive for 1, 2, 4, 7, 14 and 21 days. Activated macrophages were detected by ED1 immunostaining and quantified bilaterally in the Lumbar DRGs (L3 and L5) using image analysis system (NIS Elements). Neuropathic pain expressed by mechanical allodynia was measured using the dynamic plantar esthesiometer. The data differences were evaluated by Mann-Whitney test using STATISTICA software. Results Activated macrophages were found bilaterally. On the ipsilateral side, we observed robust increase of activated macrophages during the first four days and 21 days after CFA application. However, 7 and 14 days after was decreased number of activated macrophages. Contralateral Ganglia displayed increased number of ED1+ macrophages during the first two days, followed by a decline at 14 day after immunization with subsequent increase. A greater increase of activated macrophage was observed in L3 than L5 DRG that could be related to the afferent innervation of the ventral and dorsal side of knee joint capsule, respectively. Mechanical allodynia indicating neuropathic pain in the ipsilateral hind paws was decreased from 1 day to 14 days after RA induction. Contralateral hind paws displayed also statistically significant changes of withdrawal thresholds for mechanical allodynia from 7 to 14 days after RA induction. Conclusions Dynamic of macrophage activation was observed bilaterally in L3 and L5 DRGs of rats subjected by unilateral AIA from 1 to 21 days. The results of macrophage activation in L3 and L5 DRGs are related with afferent innervation of the knee joint. References Segond von Banchet, Gisela, et al. Experimental arthritis causes tumor necrosis factor-α-dependent infiltration of macrophages into rat dorsal root Ganglia which correlates with pain-related behavior. Pain, 2009, 145.1: 151-159. Kinne, Raimund W., et al. Macrophages in rheumatoid arthritis. Arthritis research, 2000, 2.3: 189-202. Acknowledgements Supported by the project CEITEC (CZ.1.05/1.1.00/02.0068) from European Regional Development Fund. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.2912
