Up-regulation of the TrkB receptor in mice injured by the partial ligation of the sciatic nerve

doi:10.1016/S0014-2999(00)00449-0

European Journal of Pharmacology

Volume 401, Issue 2, 4 August 2000, Pages 187-190

https://doi.org/10.1016/S0014-2999(00)00449-0 Get rights and content

Abstract

Partial nerve injury induced by tying a tight ligature around the sciatic nerve induced a marked hyperalgesia, and this persistent painful state lasted for 14 days in mice. Under these conditions, the nerve injury induced a significant increase in protein level of protein kinase Cγ isoform in plasma membranes in the spinal cord. We report here for the first time that protein level of TrkB receptor located in plasma membranes was clearly up-regulated in the spinal cord obtained from the nerve-injured mice. These findings suggest that the up-regulation of protein kinase Cγ associated with activated TrkB receptors following partial sciatic nerve ligation may induce sensitization of synaptic transmission and may in turn cause the persistent pain in mice.

Introduction

The family of neurotrophic factors collectively termed the neurotrophins includes nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3, neurotrophin-4/5 and neurotrophin-6 (Thoenen, 1991). The neurotrophins have been shown to play important roles in the regulation of persistent pain Lewin and Mendell, 1993, Theodosiou et al., 1999. The evidence to date suggests that BDNF, which is an endogenous neurotransmitter/neuromodulator in small-diameter nociceptive neurons, may be released in the spinal cord in an activity-dependent fashion, thereby regulating post-synaptic excitability within the spinal dorsal horn Theodosiou et al., 1999, Thompson et al., 1999.

The Trk family (TrkA, TrkB and TrkC) of receptor tyrosine kinases constitutes high-affinity receptors for the neurotrophins. BDNF has been shown to possess higher affinity for the TrkB receptor than TrkA and TrkC receptors (Maness et al., 1994). Recent work on inflammatory hyperalgesia has shown that behavioral nociceptive responses in inflamed rats evoked by the injection of formalin into the hind paw are reduced by intrathecal administration of TrkB-immunoglobulin G (IgG) (Thompson et al., 1999), indicating the implication of the BDNF-TrkB pathway in the inflammatory hyperalgesia.

Injury to a peripheral nerve often results in a persistent neuropathic pain condition that is characterized by spontaneous pain, allodynia and hyperalgesia. There is considerable evidence that protein kinase C contributes to the development of the long-term changes that underlie injury-associated allodynia and hyperalgesia Coderre, 1992, Mao et al., 1993, Meller et al., 1996. A recent report using the transgenic mice has provided direct evidence that activation of the gamma isoform of protein kinase is critical for the development of neuropathic pain (Malmberg et al., 1997). With respect to the intracellular mechanisms of the BDNF-TrkB pathway, the stimulation of TrkB receptor by BDNF leads to the activation of phospholipase Cγ as well as tyrosine kinases and subsequently activates protein kinase C Kaplan and Stephens, 1994, Stephens et al., 1994. Thus, these findings suggest the possibility that functional changes in TrkB receptors are implicated in the long-term hyperalgesia associated with activation of protein kinase Cγ after nerve injury. The present study was then designed to investigate the change in protein levels of the TrkB receptor and protein kinase Cγ isoform in the spinal cord following partial sciatic nerve ligation in the mouse.

Section snippets

Materials and methods

The present study was conducted in accordance with the Guiding Principles for the Care and Use of Laboratory Animals, Hoshi University, as adopted by the Committee on Animal Research of Hoshi University, which is accredited by the Ministry of Education, Science, Sports and Culture of Japan.

Results

The hyperalgesic response after the nerve injury is shown in Fig. 1. The withdrawal latencies of the ipsilateral paw to the heat thermal stimulus were significantly decreased at 7 days after the partial ligation of the sciatic nerve. The hyperalgesia lasted, at least, for 14 days after the ligation (Fig. 1). Paw withdrawal latencies in sham-operated mice on the contrary were not changed.

The changes in protein levels of protein kinase Cγ isoform and TrkB receptor after the nerve injury are

Discussion

In the present study, mice with the sciatic nerve injury displayed a marked hyperalgesia, and this persistent painful state lasted for 14 days. The present study confirmed that the nerve injury induced a significant increase in protein level of protein kinase Cγ isoform in plasma membranes in the spinal cord, as reported by Malmberg et al. (1997). To further ascertain the mechanisms that underlie the development of this persistent pain, the level of TrkB receptors after the nerve injury was

Acknowledgements

This work was supported in part by grants from the Ministry of Health and Welfare, the Ministry of Education, Science, Sports and Culture of Japan to T. Suzuki and by a research grant from the Sasakawa Health Science Foundation. We wish to thank Mr. Masahiro Shimamura and Ms. Nozomi Matsumoto for their excellent technical assistance.

References (14)

T.J. Coderre
Contribution of protein kinase C to central sensitization and persistent pain following tissue injury
Neurosci. Lett.
(1992)
G.R Lewin et al.
Nerve growth factor and nociception
Trends Neurosci.
(1993)
L.M. Maness et al.
The neurotrophines and their receptors: structure, function, and neuropathology
Neuro Biobehav. Res.
(1994)
S.T. Meller et al.
Acute thermal hyperalgesia in the rat is produced by activation of N-methyl-d-aspartate receptors and protein kinase C and production of nitric oxide
Neuroscience
(1996)
Z. Seltzer et al.
A novel behavioral model of neuropathic pain disorders produced in rats by partial sciatic nerve injury
Pain
(1990)
R.M. Stephens et al.
Trk receptors use redundant signal transduction pathways involving SHC and PLC-γ1 to mediate NGF responses
Neuron
(1994)
M. Theodosiou et al.
Hyperalgesia due to nerve damage: role of nerve growth factor
Pain
(1999)

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

Cited by (53)

Brain-derived neurotrophic factor (BDNF) in the rostral anterior cingulate cortex (rACC) contributes to neuropathic spontaneous pain-related aversion via NR2B receptors
2016, Brain Research Bulletin
Citation Excerpt :
Geng and Leal reported that the BDNF-mediated signaling pathway in the spinal cord contributes to the induction of neuropathic pain in spinal nerve ligated (SNL) rats (Geng et al., 2010; Leal et al., 2014). TrkB, a high affinity receptor of BDNF, was also markedly upregulated in mice with peripheral nerve injury caused by a partial sciatic nerve ligation (Narita et al., 2000). Thibault et al. recently found increased BDNF expression in the ACC of rats with inflammation, while local injection of a TrkB receptor antagonist mostly alleviated neuronal hyperexcitability and prevented passive avoidance behavior, confirming that the BDNF/TrkB signaling pathway in the ACC was a tuner of the affective aspect of inflammatory pain (Thibault et al., 2014).
The rostral anterior cingulate cortex (rACC) plays an important role in pain affect. Previous investigations have reported that the rACC mediates the negative affective component of inflammatory pain and contributed to the aversive state of nerve injury-induced neuropathic pain. Brain-derived neurotrophic factor (BDNF), an activity-dependent neuromodulator in the adult brain, is believed to play a role in the development and maintenance of inflammatory and neuropathic pain in the spinal cord. However, whether and how BDNF in the rACC regulates pain-related aversion due to peripheral nerve injury is largely unknown. Behaviorally, using conditioned place preference (CPP) training in rats, which is thought to reveal spontaneous pain-related aversion, we found that CPP was acquired following spinal clonidine in rats with partial sciatic nerve transection. Importantly, BDNF was upregulated within the rACC in of rats with nerve injury and enhanced the CPP acquisition, while a local injection of a BDNF-tropomyosin receptor kinase B (TrkB) antagonist into the rACC completely blocked this process. Finally, we demonstrated that the BDNF/TrkB pathway exerted its function by activating the NR2B receptor, which is widely accepted to be a crucial factor contributing to pain affect. In conclusion, our results demonstrate that the BDNF/TrkB-mediated signaling pathway in the rACC is involved in the development of neuropathic spontaneous pain-related aversion and that this process is dependent upon activation of NR2B receptors. These findings suggest that suppression of the BDNF-related signaling pathway in the rACC may provide a novel strategy to overcome pain-related aversion.
Contribution of the spinal cord BDNF to the development of neuropathic pain by activation of the NR2B-containing NMDA receptors in rats with spinal nerve ligation
2010, Experimental Neurology
The NMDA receptor and the brain-derived neurotrophic factor (BDNF) are involved in central sensitization and synaptic plasticity in the spinal cord. To determine whether the spinal cord BDNF contributes to the development and maintenance of neuropathic pain by activation of the dorsal horn NR2B-containing NMDA (NMDA-2B) receptors, this study was designed to investigate if alterations in BDNF and its TrkB receptor in the spinal dorsal horn would parallel the timeline of the development of neuropathic pain in lumbar 5 (L5) spinal nerve ligated (SNL) rats. The enzyme-linked immunosorbent assay (ELISA) showed that the BDNF concentration significantly increased during 24 h post-surgery, and the maximal enhancement lasted for 48 h. It declined as time progressed and returned to the level of pre-operation at 28 days after SNL. In parallel with the alteration of BDNF concentration in the spinal dorsal horn, the 50% paw withdrawal threshold (PWT) of the ipsilateral hind paw in SNL rats also showed a significant decrease during 24–48 h after SNL as compared with those in sham-operated rats. The correlation analysis revealed that the BDNF concentration had a negative correlation with 50% PWT in early stage (0–48 h) (r = -0.974, p = 0.001), but not late stage (3–28 days) (r = 0.3395, p = 0.6605), after SNL. Similarly, the immunohistochemical staining revealed that a significant up-regulation of BDNF expression in the spinal dorsal horn appeared as early as 12 h post-operation in SNL rats, peaked at 24–48 h, declined at 3 days and disappeared at 14 days after SNL. In contrast, an increase in NMDA-2B receptors expression in the spinal dorsal horn was delayed to 48 h after SNL. The increase reached peak at 3 days, lasted for 14 days, and returned to the control level of pre-operation at 28 days after SNL. The maximal enhancement of BDNF expression occurred in early stage (24–48 h) after nerve injury, while the peak of NMDA-2B receptors expression appeared in late stage (3–14 days) post-nerve ligation. As compared with the dynamic changes of 50% PWT in the timeline after nerve injury, the maximal enhancement of BDNF expression closely paralleled the maximal decline in the slope of 50% PWT, while the peak of NMDA-2B receptors expression corresponded with the plateau of the decreased 50% PWT. Therefore, the increased BDNF in the spinal dorsal horn was likely to be associated with the initiation of neuropathic pain in early stage (0–48 h), while the activation of NMDA-2B receptors was involved in the maintenance of persistent pain states in late stage (2–14 days) after nerve injury. Moreover, the present study also demonstrated that the BDNF/TrkB-mediated signaling pathway within the spinal cord might be involved in the induction of neuropathic pain in early stage after nerve injury, and the selective NMDA-2B receptors antagonist (Ro 25-6981) almost completely blocked the BDNF-induced mechanical allodynia in all of the tested rats. These data suggested that the BDNF/TrkB-mediated signaling pathway in the spinal cord was involved in the development of nerve injury-induced neuropathic pain through the activation of dorsal horn NMDA-2B receptors.
Chapter 19 New Therapy for Neuropathic Pain
2009, International Review of Neurobiology
Citation Excerpt :
One is activation of N‐methyl‐d‐aspartate (NMDA) receptors in postsynaptic cells (Mao et al., 1995a,b). The second mechanism is an autophosphorylation of TrkB receptor by brain‐derived neurotrophic factor (BDNF) (Narita et al., 2000; Yajima et al., 2005). In fact, the development of the hyperalgesia and allodynia in neuropathic pain states is suppressed by administration of NMDA receptor antagonists, TrkB/Fc chimera protein (sequesters endogenous BDNF), or protein kinase C inhibitors (Mao et al., 1992, 1995a; Yajima et al., 2005).
Neuropathic pain is one of the worst painful symptoms in clinic. It contains nerve‐injured neuropathy, diabetic neuropathy, chronic inflammatory pain, cancer pain, and postherpes pain, and is characterized by a tactile allodynia and hyperalgesia. Neuropathic pain, especially the nerve‐injured neuropathy, the diabetic neuropathy, and the cancer pain, is opioid resistant pain. Since the downregulation of μ‐opioid receptors is observed in dorsal spinal cord, morphine and fentanyl could not provide marked antihyperalgesic/antiallodynic effects in the course neuropathic pain states. The downregulation of μ‐opioid receptors is suggested to be mediated through the activation of NMDA receptors. Moreover, at the neuropathic pain states, the increased expression of voltage‐dependent Na⁺ channels and Ca²⁺ channels are observed. Based on the above information concerned with the pathophysiology of neural changes in neuropathic pain states, new drug treatments for neuropathic pain, using ketamine, methadone, and gabapentin, have been developed. These drugs show remarkable effectiveness against hyperalgesia and allodynia during neuropathic pain states. Oxycodone is a μ‐opioid receptor agonist, which has different pharmacological profiles with morphine. The remarkable effectiveness of oxycodone for neuropathic pain provides the possibility that μ‐opioid receptor agonists, which have different pharmacological profile with morphine, can be used for the management of neuropathic pain.
BDNF as a pain modulator
2008, Progress in Neurobiology
At least some neurotrophins may be powerful modulators of synapses, thereby influencing short- and long-term synaptic efficiency. BDNF acts at central synapses in pain pathways both at spinal and supraspinal levels. Neuronal synthesis, subcellular storage/co-storage and release of BDNF at these synapses have been characterized on anatomical and physiological grounds, in parallel with trkB (the high affinity BDNF receptor) distribution. Histological and functional evidence has been provided, mainly from studies on acute slices and intact animals, that BDNF modulates fast excitatory (glutamatergic) and inhibitory (GABAergic/glycinergic) signals, as well as slow peptidergic neurotrasmission in spinal cord. Recent studies have unraveled some of the neuronal circuitries and mechanisms involved, highlighting the key role of synaptic glomeruli in lamina II as the main sites for such a modulation.
Role of spinal voltage-dependent calcium channel α<inf>2</inf>δ-1 subunit in the expression of a neuropathic pain-like state in mice
2007, Life Sciences
The present study was undertaken to investigate the role of spinal voltage-dependent calcium channel α₂δ-1 subunit in the expression of a neuropathic pain-like state induced by partial sciatic nerve ligation in mice. In cultured spinal neurons, gabapentin (GBP), which displays the inhibitory effect of α₂δ-1 subunit, suppressed the extracellular Ca²⁺ influx induced by KCl, whereas it failed to inhibit the intracellular Ca²⁺ release induced by inositol-1,4,5-triphosphate. Seven days after sciatic nerve ligation, the protein level of α₂δ-1 subunit in the ipsilateral spinal cord was clearly increased compared to that observed in sham-operated mice. In addition, the mRNA level of α₂δ-1 subunit was significantly increased in the dorsal root ganglion, but not in the spinal cord, of nerve-ligated mice. Under these conditions, a marked decrease in the latency of paw-withdrawal against a thermal stimulation and tactile stimulation, induced by sciatic nerve ligation was abolished by repeated intrathecal (i.t.) treatment with GBP. Additionally, the persistent reduction in the nociceptive threshold by i.t. treatment with GBP at the early stage of the neuropathic pain-like state was maintained for 7 days even after GBP withdrawal. It is of interest to note that a single i.t. post-injection of GBP showed a marked and transient inhibitory effect on the developed neuropathic pain-like state, whereas repeated i.t. post-treatment with GBP produced a persistent inhibitory effect during the treatment. In conclusion, we propose here that the neuropathic pain-like state with sciatic nerve ligation is associated with the increased level of the α₂δ-1 subunit of Ca²⁺ channels at the sensory nerve terminal in the spinal dorsal horn of mice. Furthermore, the present data provide evidence that the neuropathic pain may be effectively controlled by repeated treatment with GBP at the early stage.
Involvement of microglia in the ethanol-induced neuropathic pain-like state in the rat
2007, Neuroscience Letters
Central mechanisms of neuropathy induced by chronic ethanol treatment are almost unknown. In this study, rats were treated with ethanol-diet for 72 days. Mechanical hyperalgesia was observed during ethanol consumption, even after ethanol withdrawal. Under these conditions, a microglial marker ionized calcium-binding adaptor molecule 1-, but not a neuron marker microtuble associated protein-2-, like immunoreactivies were increased in the rat spinal cord. Furthermore, hypertrophy of microglia was clearly observed following chronic ethanol consumption. These findings support the idea that the activation and hypertrophy of microglia in the spinal cord may be, at least in part, associated with in the induction of ethanol-dependent neuropathic pain-like state.

View all citing articles on Scopus

View full text

Short communicationUp-regulation of the TrkB receptor in mice injured by the partial ligation of the sciatic nerve

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Acknowledgements

Neurosci. Lett.

Trends Neurosci.

Neuro Biobehav. Res.

Neuroscience

Pain

Neuron

Pain

Short communication
Up-regulation of the TrkB receptor in mice injured by the partial ligation of the sciatic nerve