Tomosada Sugimoto

Activation of microglia and p38 mitogen-activated protein kinase in the dorsal column nucleus contributes to tactile allodynia following peripheral nerve injury.

The activation of glial cells in the CNS has been suggested to be involved in abnormal pain sensation after peripheral nerve injury. Previous studies demonstrated phosphorylation of p38 mitogen-activated protein kinase (MAPK) in spinal cord glial cells after peripheral nerve injury, and such phosphorylation has been suggested to be involved in the development of neuropathic pain. The aim of this study was to examine the dorsal column nuclei for phosphorylation of p38 MAPK following peripheral nerve injury and to explore a possibility of its contribution to neuropathic pain. Immunohistochemical labeling for phosphorylated p38 (p-p38) MAPK was performed in histological sections of the rat spinal cord and medulla oblongata after the fifth lumbar (L5) spinal nerve ligation (SNL). The number of p-p38 MAPK-immunoreactive (IR) cells was significantly increased in the L5 dorsal horn and the gracile nucleus ipsilateral to the injury at days 3-21 after SNL. Double immunofluorescence labeling with cell-specific markers revealed that p-p38 MAPK-IR cells co-expressed OX-42, suggesting their microglial identity. Increased immunofluorescence labeling for OX-42 indicated that microglial cells were activated by SNL in the L5 dorsal horn and the gracile nucleus ipsilateral to the injury. Continuous infusion of a p38 MAPK inhibitor into the cisterna magna for 14 days beginning on the day of SNL suppressed the development of tactile allodynia, but not thermal hyperalgesia induced by nerve injury. These results demonstrate that SNL activates p38 MAPK pathway in microglia in the gracile nucleus as well as in the spinal cord dorsal horn. Activation of p38 MAPK in medullary microglia may contribute to the pathogenesis of neuropathic pain.

Differential activation of mitogen-activated protein kinases and glial cells in the trigeminal sensory nuclear complex following lingual nerve injury

Mitogen-activated protein kinases (MAPKs) play a pivotal role in the mediation of cellular responses to a variety of signaling molecules. The current study demonstrates phosphorylation of extracellular signal-regulated kinase (ERK) and p38 MAPK in each subdivision of the trigeminal sensory nuclear complex (TSNC) following lingual nerve injury. Immunohistochemical labeling for phosphorylated ERK (p-ERK) or phosphorylated p38 (p-p38) MAPK was performed in histological sections of the brainstem. A transient increase in the immunoreactivity for p-ERK was found in each subdivision of the TSNC followed by a prolonged increase in the immunoreactivity for p-p38 MAPK after nerve injury. Double immunofluorescence labeling with cell-specific markers revealed that ERK and p38 MAPK were phosphorylated predominantly by OX-42-positive microglia or GFAP-positive astrocytes. Increased immunofluorescence labeling for OX-42 and GFAP indicated that microglia and astrocytes were activated by nerve injury in the TSNC. Activation of MAPKs and glial cells in the rostral subdivisions of the TSNC was comparable with that in the subnucleus caudalis of the trigeminal spinal tract nucleus (Vc). We conclude that differential activation of MAPKs and glial cells in the rostral subdivisions of the TSNC as well as the Vc may have a substantial role in the pathogenesis of neuropathic pain following trigeminal nerve injury.

Effect of adenosine A1 receptor agonist on the enhanced excitability of spinal dorsal horn neurons after peripheral nerve injury

Neuronal hyperactivity has been implicated in abnormal pain sensation following peripheral nerve injuries. Previous studies have indicated that the activation of adenosine A1 receptors (A1R) in the central and peripheral nervous systems produces an antinociceptive effect. However, the mechanisms involved in the peripheral effect are still not fully understood. The effects of the local application of the selective A1R agonist, 2-chloro-N(6)-cyclopentyladenosine (CCPA) on neuronal hyperactivity were examined in this study using a neuropathic pain model induced by a tibial nerve injury. We utilized Fos protein-like immunoreactivity induced by noxious heat stimulation to examine changes in the number of Fos protein like immunoreactive (Fos-LI) neuron profiles in the spinal dorsal horn, and behavioral analysis for mechanical and thermal sensitivities. The nerve injury induced an exaggerated Fos response to noxious heat stimulation. The number of Fos-LI neuron profiles was significantly decreased and their distribution was restricted to the central terminal field of the spared peroneal nerve 3 days after the injury. The number of Fos-LI neuron profiles returned to control levels and a large number of these profiles were observed in the central terminal field of the injured tibial nerve 14 days after the injury. These enhanced Fos responses were attenuated by the local application of CCPA. The nerve injury also resulted in mechanical allodynia and thermal hyperalgesia. The local application of CCPA inhibited thermal hyperalgesia, but was less effective against mechanical allodynia. These results indicated that activation of peripheral A1R plays a role in the regulation of nerve injury-induced hyperalgesia.

Cross-Excitation in Peripheral Sensory Ganglia Associated with Pain Transmission

Despite the absence of synaptic contacts, cross-excitation of neurons in sensory ganglia during signal transmission is considered to be chemically mediated and appears increased in chronic pain states. In this study, we modulated neurotransmitter release in sensory neurons by direct application of type A botulinum neurotoxin (BoNT/A) to sensory ganglia in an animal model of neuropathic pain and evaluated the effect of this treatment on nocifensive. Unilateral sciatic nerve entrapment (SNE) reduced the ipsilateral hindpaw withdrawal threshold to mechanical stimulation and reduced hindpaw withdrawal latency to thermal stimulation. Direct application of BoNT/A to the ipsilateral L4 dorsal root ganglion (DRG) was localized in the cell bodies of the DRG and reversed the SNE-induced decreases in withdrawal thresholds within 2 days of BoNT/A administration. Results from this study suggest that neurotransmitter release within sensory ganglia is involved in the regulation of pain-related signal transmission.

Persistent hypoglossal artery with hypoplasia of the vertebral and posterior communicating arteries

We observed a rare case of the right persistent hypoglossal artery (PHA) in the posterior cranial fossa of a deceased 74-year-old Japanese man who did not have any clinical manifestations related to this anomaly when alive. The large-sized PHA originating from the cervical internal carotid artery passed through the hypoglossal canal together with the hypoglossal nerve and reached the posterior cranial fossa to anastomose the basilar artery. In addition, the ipsilateral vertebral artery and bilateral posterior communicating arteries were hypoplastic. Here, we discuss the developmental mechanisms underlying the formation of the PHA and the spectrum of diseases related to its presence.

The number of nociceptors in the trigeminal ganglion but not proprioceptors in the mesencephalic trigeminal tract nucleus is reduced in dystonin deficient dystonia musculorum mice

The trigeminal ganglion (TG) and mesencephalic trigeminal tract nucleus (Mes5) were investigated in wild type and dystonia musculorum (dt) mice to study the effect of dystonin deficiency on primary sensory neurons in the trigeminal nervous system. At postnatal day 14, the number of TG neurons was markedly decreased in dt mice when compared to wild type mice (43.1% reduction). In addition, dystonin disruption decreased the number of sensory neurons which bound to isolectin B4, and contained calcitonin gene-related peptide or high-affinity nerve growth factor receptor TrkA. Immunohistochemistry for caspase-3 demonstrated that dystonin deficiency induced excess cell death of TG neurons during the early postnatal period. In contrast, Mes5 neurons were barely affected in dt mice. These data together suggest that dystonin is necessary for survival of nociceptors but not proprioceptors in the trigeminal nervous system.

The reduction of proprioceptors in the mesencephalic trigeminal tract nucleus after neonatal masseteric nerve transection; effect of brain-derived neurotrophic factor.

The effect of neonatal masseteric nerve transection on primary proprioceptors was examined in the mesencephalic trigeminal tract nucleus (Mes5) of the rat. At 72 h to 21 days after the injury, the number of Mes5 neurons decreased on the side ipsilateral to the transection. The means+/-SD of percentage proportion of ipsilateral/contralateral neurons at 72 h and 21 days were 69.9+/-7.5% and 58.2+/-14.6%, respectively. The application of brain-derived neurotrophic factor to the proximal stump of the masseteric nerve delayed the loss of Mes5 neurons at 72 h after the injury; the mean numbers+/-SD of ipsilateral and contralateral Mes5 neurons in injured animals with BDNF application was 553.6+/-61.9 and 558.4+/-55.3, respectively. Saline application had no effect on the injury-induced loss of Mes5 neurons; i.e., the mean numbers+/-SD of ipsilateral and contralateral Mes5 neurons were 367.3+/-72.5 and 543+/-33.5, respectively. These findings indicate that trigeminal primary proprioceptors are sensitive to the neonatal injury. The survival of proprioceptors during early postnatal period is probably dependent upon brain-derived neurotrophic factor in the trigeminal nervous system.

PEPTIDE 19 IN THE RAT SUPERIOR CERVICAL GANGLION

Peptide 19 is a 7.6 kDa polypeptide which can bind to calmodulin and inhibit calcium-calmodulin signaling. In this study, peptide 19-immunoreactivity was examined in the rat superior cervical ganglion. In the ganglion, 54.8% of postganglionic sympathetic neuron profiles were immunoreactive for peptide 19. These neuron profiles were small- to medium-sized and measured 87-845 microm(2) (mean+/-SD = 343+/-111 microm(2)). Double immunofluorescence method revealed that 99.9% of peptide 19-containing neurons had neuropeptide Y in the superior cervical ganglion. Retrograde neuronal tracing and immunohistochemical studies also demonstrated that peptide 19 was common in postganglionic sympathetic neurons which innervated the facial skin and masseter but not the submandibular gland; 55.6% and 75.2% of cutaneous and muscular neuron profiles, respectively, contained peptide 19. Only 9.8% of glandular neurons were immunoreactive for peptide 19. These findings indicate that the content of peptide 19 in superior cervical ganglion neurons depends on their cell sizes and peripheral projections. On the other hand, colchicine injection into the superior cervical ganglion decreased the number of peptide 19-positive neurons (30.7%) compared to saline injection (53.3%). In contrast, the treatment induced nicotine adenine dinucleotide phosphate diaphorase activity in 12.7% of postganglionic sympathetic neurons. Double stain demonstrated that 56.3% of nicotine adenine dinucleotide phosphate diaphorase-positive neurons co-expressed peptide 19. These findings indicate that colchicine treatment causes decrease of peptide 19 expression and increase of nitric oxide synthase activity.

Assessment of intraoral mucosal pain induced by the application of capsaicin

OBJECTIVE To develop an objective method for assessing nociceptive behaviour in an animal model of capsaicin-induced intraoral pain. Changes in nociceptive responses were also examined after injury to the inferior alveolar nerve (IAN). DESIGN Nociceptive responses evoked by the intraoral application of various doses of capsaicin were analyzed in lightly anaesthetized rats. The number of c-Fos protein-like immunoreactive (Fos-LI) neurons in the medullary dorsal horn (MDH) induced by the intraoral application of capsaicin was measured. Behavioural and c-Fos responses were also examined 14 days after injury to the IAN. RESULTS Larger doses of intraoral capsaicin (1, 10 and 100μg) induced vigorous licking behaviour and c-Fos response in the MDH in a reproducible manner. The magnitudes of both behavioural activity and the c-Fos response from the 10 and 100μg doses of capsaicin were significantly greater than that by the 1μg dose. Injury to the IAN exaggerated the behavioural and c-Fos responses evoked by intraoral capsaicin. CONCLUSIONS The intraoral application of capsaicin is a valid and reliable method for studying intraoral pain and hyperalgesia following nerve injury.

Differential induction of c-Fos and phosphorylated ERK by a noxious stimulus after peripheral nerve injury

ABSTRACT Purpose: In this study, we compared induction of c-Fos and phosphorylated extracellular signal-regulated kinase (p-ERK) in the spinal dorsal horn after peripheral nerve injury. Materials and methods: We examined the spinal dorsal horn for noxious heat-induced c-Fos and p-ERK protein-like immunoreactive (c-Fos- and p-ERK-IR) neuron profiles after tibial nerve injury. The effect of administration of a MEK 1/2 inhibitor (PD98059) on noxious heat-induced c-Fos expression was also examined after tibial nerve injury. Results: A large number of c-Fos- and p-ERK-IR neuron profiles were induced by noxious heat stimulation to the hindpaw in sham-operated animals. A marked reduction in the number of c-Fos- and p-ERK-IR neuron profiles was observed in the medial 1/3 (tibial territory) of the dorsal horn at 3 and 7 days after nerve injury. Although c-Fos-IR neuron profiles had reappeared by 14 days after injury, the number of p-ERK-IR neuron profiles remained decreased in the tibial territory of the superficial dorsal horn. Double immunofluorescence labeling for c-Fos and p-ERK induced by noxious heat stimulation to the hindpaw at different time points revealed that a large number of c-Fos-IR, but not p-ERK-IR, neuron profiles were distributed in the tibial territory after injury. Although administration of a MEK 1/2 inhibitor to the spinal cord suppressed noxious heat-induced c-Fos expression in the peroneal territory, this treatment did not alter c-Fos induction in the tibial territory after nerve injury. Conclusions: ERK phosphorylation may be involved in c-Fos induction in normal nociceptive responses, but not in exaggerated c-Fos induction after nerve injury.