The proportion of TRPV1 protein-positive lumbar DRG neurones does not increase in the course of acute and chronic antigen-induced arthritis in the knee joint of the rat

doi:10.1016/j.neulet.2003.12.034

Neuroscience Letters

Volume 361, Issues 1–3, 6 May 2004, Pages 172-175

https://doi.org/10.1016/j.neulet.2003.12.034 Get rights and content

Abstract

The TRPV1 receptor, previously called VR1 receptor, is a non-selective cation channel gated by capsaicin, noxious heat, protons and anandamide. The TRPV1 receptor is essential for the development of thermal hyperalgesia. The present study investigated whether the proportion of neurones with TRPV1 receptor increases in lumbar DRG neurones in the course of an antigen-induced arthritis (AIA) of one knee joint in the rat. In control rats 38.1±2.3% of the neurones from sections of the L1–L5 ganglia showed TRPV1-like immunoreactivity. Neither in the acute (3 days) nor chronic phase (21 days) of AIA in the knee joint the proportion of TRPV1-like immunoreactive profiles showed significant changes. Thus AIA in the knee joint is not associated with an up-regulation of the TRPV1 receptor in the lumbar DRG neurones.

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Acknowledgements

The authors thank Mrs U. Möller for assistance in the immunohistochemistry and Mrs R. Stöckigt for here excellent technical assistance.

References (16)

F. Amaya et al.
Local inflammation increases vanilloid receptor 1 expression within distinct subgroups of DRG neurons
Brain Res.
(2003)
R. Bron et al.
Activation of ras is necessary and sufficient for upregulation of vanilloid receptor type 1 in sensory neurons by neurotrophic factors
Mol. Cell. Neurosci.
(2003)
E. Buchner et al.
Induction of flare-up reactions in rat antigen-induced arthritis
J. Autoimmun.
(1995)
S.M. Carlton et al.
Peripheral capsaicin receptors increase in the inflamed rat hindpaw: a possible mechanism for peripheral sensitization
Neurosci. Lett.
(2001)
T. Fukuoka et al.
VR1, but not P2X3, increases in the spared L4 DRG in rats with L5 spinal nerve ligation
Pain
(2002)
K.J. Halbhuber et al.
Modern laser scanning microscopy in biology, biotechnology and medicine
Ann. Anatomy
(2003)
X. He et al.
Capsaicin inhibits responses of fine afferents from the knee joint of the cat to mechanical and chemical stimuli
Brain Res.
(1990)
R.-R. Ji et al.
p38 MAPK activation by NGF in primary sensory neurons after inflammation increases TRPV1 levels and maintains heat hyperalgesia
Neuron
(2002)

There are more references available in the full text version of this article.

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Amelioration of experimentally induced inflammatory arthritis by intra-articular injection of visnagin
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Citation Excerpt :
All efforts were made to minimize animals suffering. Edema of the knee joint of animals was measured as the symbol of disease index using digital Vernier Caliper (Bär et al., 2004). Briefly, hairs on the knee joints of the animals were shaved before experimentation and width of the knee was measured on days 0, 7, 14, 21 of the study period.
Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease characterized by synovial hyperplasia, cartilage destruction and bone erosion. Visnagin (VIS) is a proven anti-inflammatory agent and in this study, we aimed to evaluate the anti-arthritic activity of VIS when administered via intra-articular (I.A.) route of administration.
RAW 264.7 cells were stimulated with lipopolysaccharide (LPS) (1 μg/mL) and treated with VIS at concentrations of 12.5 and 25 μM. Arthritis was induced in Sprague Dawley rats by administering Complete Freund's Adjuvant (CFA) (1 mg/mL) through (I.A.) route and treated with VIS via (I.A.) route at doses of 3 and 10 mg/kg twice a week for 3 weeks. Protective effects were assessed by arthritic score, behavioral studies for pain evaluation, radiological assessment, histopathological examination and molecular studies.
Our results indicated that VIS significantly reduced LPS induced inflammation in RAW 264.7 cells. While in arthritic rats, VIS reduced the disease scorings with improvement towards pain. Pathological examination demonstrated that VIS reduced knee joint inflammation and cartilage destruction. Radiographic analysis and molecular studies also supported the protective effects of VIS.
The results of the study imply that VIS exerted potential anti-inflammatory and anti-arthritic activity in in vitro and in vivo models of RA.
The potential anti-inflammatory and anti-nociceptive effects of rat hemopressin (PVNFKFLSH) in experimental arthritis
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Citation Excerpt :
All the experiments were blinded to the main investigators. The induction of AIA was performed as previously described (Andersson et al., 1998; Bär et al., 2004). Briefly, rats were temporarily anaesthetized with halothane (1.5% vol/vol in oxygen; Takaoka, Mod KT-20, São Paulo, Brazil) and subsequently they were immunized at the base of the tail with a subcutaneous (s.c.) injection of 50 μL of the antigen: 40 mg/ml of methylated bovine serum albumin (m-BSA) in 5% glucose solution emulsified with Freund's complete adjuvant (1:1, v/v).
Inflammatory arthritis, such as rheumatoid arthritis (RA), stands out as one of the main sources of pain and impairment to the quality of life. The use of hemopressin (PVNFKFLSH; Hp), an inverse agonist of type 1 cannabinoid receptor, has proven to be effective in producing analgesia in pain models, but its effect on neuro-inflammatory aspects of RA is limited. In this study, antigen-induced arthritis (AIA) was evoked by the intraarticular (i.art.) injection of methylated bovine serum albumin (mBSA) in male Sprague Dawley rats. Phosphate buffered saline (PBS)-injected ipsilateral knee joints or AIA contralateral were used as control. Nociceptive and inflammatory parameters such as knee joint oedema and leukocyte influx and histopathological changes were carried out in addition to the local measurement of interleukins (IL) IL-6, IL-1β, tumor necrosis factor-α and the immunoreactivity of the neuropeptides substance P (SP) and calcitonin gene related peptide (CGRP) in the spinal cord (lumbar L3-5 segments) of AIA rats. For 4 days, AIA rats were treated daily with a single administration of saline, Hp injected (10 or 20 μg/day, i.art.), Hp given orally (20 μg/Kg, p.o.) or indomethacin (Indo; 5 mg/Kg, i.p.). In comparison to the PBS control group, the induction of AIA produced a significant and progressive mono-arthritis condition. The degree of AIA severity progressively compromised the normal walking pattern and impaired mobility over the next four days in relation to PBS-injected rats or contralateral knee joints. In AIA rats, the reduction of the distance between footprints and disturbances of gait evidenced signs of nociception. This response worsened at day 4, and a loss of footprint from the ipsilateral hind paw was evident. Daily treatment of the animals with Hp either i.art. (10 and 20 μg/knee) or p.o. (20 μg/Kg) as well as Indo (5 mg/Kg, i.p.) ameliorated the impaired mobility in a time-dependent manner (P < 0.05). In parallel, the AIA-injected ipsilateral knee joints reach a peak of swelling 24 h after AIA induction, which persisted over the next four days in relation to PBS-injected rats or contralateral knee joints. There was a significant but not dose-dependent inhibitory effect produced by all dosages and routes of Hp treatments on AIA-induced knee joint swelling (P < 0.05). In addition, the increased synovial levels of MPO activity, total leukocytes number and IL-6, but not IL-1β, were significantly reduced by the lower i.art. dose of Hp. In conclusion, these results successfully demonstrate that Hp may represent a novel therapeutic strategy to treat RA, an effect which is unrelated to the proinflammatory actions of the neuropeptides CGRP and SP.
Acute inflammation sensitizes knee-innervating sensory neurons and decreases mouse digging behavior in a TRPV1-dependent manner
2018, Neuropharmacology
Citation Excerpt :
Some studies report that TRPV1−/− mice do not develop CFA- (Keeble et al., 2005) or carrageenan- (Davis et al., 2000) induced hyperalgesia and others have shown that in DRG neurons TRPV1 protein expression increases in CFA-induced inflammation (Amaya et al., 2003; Ji et al., 2002; Yu et al., 2008). In direct contrast, other studies failed to show increased neuronal TRPV1 protein (Bär et al., 2004; Zhou et al., 2003) or mRNA expression (Ji et al., 2002) following CFA-induced inflammation. Similarly, TRPA1−/− mice show normal development of CFA-induced hyperalgesia in some studies (Fernandes et al., 2011; Petrus et al., 2007), whereas others report attenuation of CFA- and carrageenan-induced hyperalgesia in TRPA1−/− mice (Garrison and Stucky, 2014; Moilanen et al., 2012) or following administration of a TRPA1 antagonist (Bonet et al., 2013; Samer R Eid et al., 2008).
Ongoing, spontaneous pain is characteristic of inflammatory joint pain and reduces an individual's quality of life. To understand the neural basis of inflammatory joint pain, we made a unilateral knee injection of complete Freund's adjuvant (CFA) in mice, which reduced their natural digging behavior. We hypothesized that sensitization of knee-innervating dorsal root ganglion (DRG) neurons underlies this altered behavior. To test this hypothesis, we performed electrophysiological recordings on retrograde labeled knee-innervating primary DRG neuron cultures and measured their responses to a number of electrical and chemical stimuli. We found that 24-h after CFA-induced knee inflammation, knee neurons show a decreased action potential generation threshold, as well as increased GABA and capsaicin sensitivity, but have unaltered acid sensitivity. The inflammation-induced sensitization of knee neurons persisted for 24-h in culture, but was not observed after 48-h in culture. Through immunohistochemistry, we showed that the increased knee neuron capsaicin sensitivity correlated with enhanced expression of the capsaicin receptor, transient receptor potential vanilloid 1 (TRPV1) in knee-innervating neurons of the CFA-injected side. We also observed an increase in the co-expression of TRPV1 with tropomyosin receptor kinase A (TrkA), which is the receptor for nerve growth factor (NGF), suggesting that NGF partially induces the increased TRPV1 expression. Lastly, we found that systemic administration of the TRPV1 antagonist, A-425619, reversed the decrease in digging behavior induced by CFA injection, further confirming the role of TRPV1, expressed by knee neurons, in acute inflammatory joint pain.
Effect of genetic deletion of the vanilloid receptor TRPV1 on the expression of Substance P in sensory neurons of mice with adjuvant-induced arthritis
2010, Neuropeptides
The neuropeptide Substance P (SP), expressed by nociceptive sensory afferents in joints, plays an important role in the pathogenesis of arthritis. Capsaicin causes neurons in the dorsal root ganglia (DRG) to release SP from their central and peripheral axons, suggesting a functional link between SP and the capsaicin receptor, the transient receptor potential vanilloid 1 (TRPV1). The expression of both TRPV1 and SP have been reported to increase in several models of arthritis but the specific involvement of TRPV1-expressing articular afferents that can release SP is not completely understood. We here wanted to ascertain whether the increase in the number of SP-positive primary afferents in arthritis may be affected by genetic deletion of TRPV1. For this, we used immunohistochemistry to quantify the expression of SP in primary afferent neurons in wild-type mice (WT) vs. TRPV1-knockout (KO) mice with adjuvant-induced arthritis (AIA). We found that the expression of SP in DRG (1) increased significantly over naïve level in both WT and KO mice 3 weeks after AIA, (2) was significantly higher in KO mice than in WT mice in naïve mice and 2–3 weeks after AIA, (3) was significantly higher on the side of AIA than on the contralateral, vehicle-injected side at all time points in WT mice, but not in KO mice, and (4) increased predominantly in small-size neurons in KO mice and in small- and medium-size neurons in WT mice. Since the size distribution of SP-positive DRG neurons in arthritic TRPV1-KO mice was not significantly different from that in naïve mice, we speculate that the increased expression of SP is unlikely to reflect recruitment of A-fiber primary afferents and that the higher expression of SP in KO mice may represent a plastic change to compensate for the missing receptor in a major sensory circuit.
Detection of endogenous and immuno-bound peroxidase - The status Quo in histochemistry
2010, Progress in Histochemistry and Cytochemistry
The discovery of synthetic dyes goes back to 1856 and launched the development of the whole chemical and pharmaceutical industry. In life sciences synthetic dyes represent indispensable tools for the microscopic and macroscopic level. Small dyes have the advantage of their easy adaptability to various measuring equipments. By way of structural modification of the chromophore portion, dye labels can be tailored that they absorb and emit light at desired wavelengths ranging from the UV to the near infrared region of the spectrum. Assisted by the development of light measuring techniques and the commercial availability of highly sensitive equipment, today luminescent labels represent most sensitive detection tools in life sciences and dominate over chromogen based techniques. However, for detection of active sites of peroxidase (PO) so far fluorescent labels have been confined to only a few substrates while a broad variety of well-established chromogenic techniques exist. This review covers fluorescent and chromogenic approaches for the permanent detection of immuno-bound and endogenous PO-activity in fixed cells and tissues. Thereby the tailoring of suitable dye labels is additionally challenged by two demands: (1) The applied dye (or its precursor) must act as enzyme substrate specifically and (2) the enzymatic impact must furnish an insoluble dye product from easy soluble starting materials in a very quick reaction. Hence it is not surprising that among PO-substrates (and enzyme substrates generally), dye conjugates represent only an exception while most of these labels represent reactive dyes or suitable precursors. Chromogenic and fluorescent approaches for the permanent labeling of enzymatic sites are compiled. Furthermore, various area-spanning PO-detection principles are discussed ranging from transmission light (TLM) and fluorescence light (FLM) microscopy (chromogenes, flourochromes, fluorescent chromogenes, chromogenes with nonlinear optical properties) to correlated transmission electron microscopy (TEM; photoconversion of specific chromogenic reaction products, electron opaque and/or osmiophilic chromogenic substrates). Also, approaches for reflectance laser microscopy (RLM), polarization microscopy (PM), and correlative TLM, FLM, and multiphoton fluorescence microscopy (MFM) are discussed.
Vanilloid receptor TRPV1-mediated phosphorylation of ERK in murine adjuvant arthritis
2009, Osteoarthritis and Cartilage
Citation Excerpt :
However, our results are consistent with the report that 2 days after injection of CFA in a mouse hindpaw, the percentage of isolectin B4 of Griffonia simplicifolia (IB4)-positive DRG neurons that responded to capsaicin or H+ increased from 24% to 80% and from 54% to 85%, respectively, in parallel with an increased number of TRPV1-immunoreactive neurons57. Increased expression of TRPV1 has been reported in DRG neurons58–61 and in peripheral nerve fibers at the site of inflammation62,63, although the changes in TRPV1 expression may depend on the particular model of arthritis, survival time, and animal species64. In the present study, we have established that activation of the ERK biochemical cascade in nociceptive neurons is mediated, at least in part, by TRPV1.
The vanilloid receptor transient receptor potential vanilloid 1 (TRPV1), expressed by sensory neurons that innervate joints, is implicated in arthritis but the mechanisms are not fully understood. One possibility is that downstream effects of activation of TRPV1 are mediated by the extracellularly-regulated kinase (ERK). ERK is phosphorylated (p-ERK) in sensory neurons in response to noxious stimuli and its inhibition has been found to be antinociceptive in several pain models. We here wanted to ascertain whether TRPV1 may contribute to the pain hypersensitivity and inflammation of arthritis via an ERK-mediated pathway.
We used a model of adjuvant-induced arthritis (AIA) of the ankle and investigated the changes in expression of p-ERK in sensory afferent neurons in dorsal root ganglia (DRG) and spinal dorsal horn of TRPV1-knockout (KO) mice, compared to wild-type (WT) mice of the same genetic background, using multiple immunofluorescence.
Two to three weeks after inducing AIA in mice, the number of neurons in DRG and spinal cord that expressed p-ERK was significantly higher on the side of AIA than on the contralateral, vehicle-injected side. The fraction of p-ERK-positive neurons in the DRG that also expressed TRPV1 was increased, indicating that activation of ERK occurred preferentially in TRPV1-positive neurons. Moreover, TRPV1-KO mice had reduced activation of ERK in sensory neurons, compared to WT mice. These changes in expression of p-ERK correlated with changes in pain behavior and joint histopathology: TRPV1-KO mice had reduced nociceptive behavior and severity of arthritis, compared to WT mice.
Our results support the idea that activation of ERK in primary afferent neurons is mediated, at least in part, by TRPV1. In the absence of TRPV1, the signs of arthralgia and histopathology in the mouse model of AIA are reduced. We conclude that TRPV1, expressed by neurons in the articular afferent pathway, contributes to the pathogenesis of arthritis via an ERK-mediated pathway.

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The proportion of TRPV1 protein-positive lumbar DRG neurones does not increase in the course of acute and chronic antigen-induced arthritis in the knee joint of the rat

Abstract

Section snippets

Acknowledgements

Brain Res.

Mol. Cell. Neurosci.

J. Autoimmun.

Neurosci. Lett.

Pain

Ann. Anatomy

Brain Res.

Neuron