RESUMO
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.
Assuntos
Hiperestesia/etiologia , Bulbo/metabolismo , Microglia/fisiologia , Doenças do Sistema Nervoso Periférico/complicações , Doenças do Sistema Nervoso Periférico/patologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Comportamento Animal , Antígeno CD11b/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/administração & dosagem , Proteína Glial Fibrilar Ácida/metabolismo , Hiperestesia/tratamento farmacológico , Imidazóis/administração & dosagem , Masculino , Limiar da Dor/efeitos dos fármacos , Doenças do Sistema Nervoso Periférico/tratamento farmacológico , Fosfopiruvato Hidratase/metabolismo , Piridinas/administração & dosagem , Ratos , Ratos Sprague-Dawley , Tempo de Reação/efeitos dos fármacos , Nervos Espinhais/patologia , Fatores de TempoRESUMO
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.
Assuntos
Proteínas do Citoesqueleto/deficiência , Proteínas do Tecido Nervoso/deficiência , Nociceptores/metabolismo , Células Receptoras Sensoriais/metabolismo , Gânglio Trigeminal/citologia , Núcleos do Trigêmeo/citologia , Animais , Proteínas de Transporte , Caspase 3/metabolismo , Distonina , Regulação da Expressão Gênica/genética , Lectinas/metabolismo , Camundongos , Camundongos Knockout , Proteínas do Tecido Nervoso/metabolismo , Receptor trkA/metabolismoRESUMO
Previous studies indicated that the expression of neuropsin, a serine protease, is induced in mature oligodendrocytes after injury to the CNS. The pathophysiology of spinal cord injury (SCI) involves primary and secondary mechanisms, the latter contributing further to permanent losses of function. To explore the role of neuropsin after SCI, histochemical and behavioral analyses were performed in wild-type (WT) and neuropsin-deficient (neuropsin(-/-)) mice using a crush injury model, a well-characterized and consistently reproducible model of SCI. In situ hybridization revealed that neuropsin mRNA expression was induced in the spinal cord white matter from WT mice after crush SCI, peaking at day 4. Neuropsin(-/-) mice showed attenuated demyelination, oligodendrocyte death, and axonal damage after SCI. Although axonal degeneration in the corticospinal tract was obvious caudal to the lesion site in both strains of mice after SCI, the number of surviving nerve fibers caudal to the lesion was significantly larger in neuropsin(-/-) mice than WT mice. Behavioral analysis revealed that the recovery at days 10-42 was significantly improved in neuropsin(-/-) mice compared with WT mice in spite of the severe initial hindlimb impairments due to SCI in both strains. These observations suggest that neuropsin is involved in the secondary phase of the pathogenesis of SCI mediated by demyelination, oligodendrocyte death, and axonal degeneration.
Assuntos
Doenças Desmielinizantes/metabolismo , Calicreínas/metabolismo , Oligodendroglia/metabolismo , Traumatismos da Medula Espinal/metabolismo , Degeneração Walleriana/metabolismo , Animais , Mapeamento Encefálico , Morte Celular/genética , Doenças Desmielinizantes/genética , Doenças Desmielinizantes/fisiopatologia , Modelos Animais de Doenças , Imunofluorescência , Regulação da Expressão Gênica/fisiologia , Hemiplegia/genética , Hemiplegia/fisiopatologia , Hibridização In Situ , Calicreínas/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fibras Nervosas Mielinizadas/metabolismo , Fibras Nervosas Mielinizadas/patologia , Regeneração Nervosa/genética , Vias Neurais/metabolismo , Vias Neurais/patologia , Vias Neurais/fisiopatologia , Oligodendroglia/patologia , RNA Mensageiro/metabolismo , Recuperação de Função Fisiológica/genética , Medula Espinal/metabolismo , Medula Espinal/patologia , Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/fisiopatologia , Degeneração Walleriana/genética , Degeneração Walleriana/fisiopatologiaRESUMO
Immunohistochemistry for brain-derived neurotrophic factor (BDNF) was performed on the rat vagal and glossopharyngeal sensory ganglia. In the jugular, petrosal and nodose ganglia, 56.1+/-5.5%, 52.4+/-9.4% and 80.0+/-3.0% of sensory neurons, respectively, were immunoreactive for BDNF. These neurons were small- to medium-sized and observed throughout the ganglia. In the solitary tract nucleus, the neuropil showed BDNF immunoreactivity. A double immunofluorescence method demonstrated that BDNF-immunoreactive neurons were also immunoreactive for calcitonin gene-related peptide (CGRP), P2X3 receptor, the capsaicin receptor (VR1) or vanilloid receptor 1-like receptor (VRL-1) in the jugular (CGRP, 43.5%; P2X3 receptor, 51.1%; VR1, 71.7%; VRL-1, 0.5%), petrosal (CGRP, 33.2%; P2X3 receptor, 58.4%; VR1, 54.2%; VRL-1, 23.3%) and nodose ganglia (CGRP, 1.8%; P2X3 receptor, 49.1%; VR1, 70.7%; VRL-1, 11.5%). The co-expression with tyrosine hydroxylase was also detected in the petrosal (2.9%) and nodose ganglia (2.2%). However, BDNF-immunoreactive neurons were devoid of parvalbumin in these ganglia. The present findings suggest that BDNF-containing vagal and glossopharyngeal sensory neurons have nociceptive and chemoreceptive functions.
Assuntos
Corpos Aórticos/fisiologia , Fator Neurotrófico Derivado do Encéfalo/fisiologia , Gânglios Sensitivos/fisiologia , Nervo Glossofaríngeo/fisiologia , Neurônios/fisiologia , Animais , Corpos Aórticos/citologia , Células Quimiorreceptoras/fisiologia , Gânglios Sensitivos/citologia , Glomo Jugular/citologia , Glomo Jugular/fisiologia , Modelos Animais , Neurônios Aferentes/fisiologia , Gânglio Nodoso/citologia , Gânglio Nodoso/fisiologia , RatosRESUMO
The anterior part of the tongue was examined in wild type and dystonia musculorum mice to assess the effect of dystonin loss on fungiform papillae. In the mutant mouse, the density of fungiform papillae and their taste buds was severely decreased when compared to wild type littermates (papilla, 67% reduction; taste bud, 77% reduction). The mutation also reduced the size of these papillae (17% reduction) and taste buds (29% reduction). In addition, immunohistochemical analysis demonstrated that the dystonin mutation reduced the number of PGP 9.5 and calbindin D28k-containing nerve fibers in fungiform papillae. These data together suggest that dystonin is required for the innervation and development of fungiform papillae and taste buds.
Assuntos
Proteínas de Transporte/genética , Proteínas do Citoesqueleto/genética , Proteínas do Tecido Nervoso/genética , Papilas Gustativas/anormalidades , Papilas Gustativas/metabolismo , Distúrbios do Paladar/metabolismo , Língua/anormalidades , Língua/metabolismo , Animais , Calbindina 1 , Calbindinas , Nervo da Corda do Tímpano/anormalidades , Nervo da Corda do Tímpano/metabolismo , Nervo da Corda do Tímpano/fisiopatologia , Modelos Animais de Doenças , Distúrbios Distônicos/genética , Distúrbios Distônicos/metabolismo , Distúrbios Distônicos/fisiopatologia , Distonina , Gânglio Geniculado/anormalidades , Gânglio Geniculado/metabolismo , Gânglio Geniculado/fisiopatologia , Imuno-Histoquímica , Camundongos , Camundongos Knockout , Mutação/genética , Proteína G de Ligação ao Cálcio S100/metabolismo , Células Receptoras Sensoriais/anormalidades , Células Receptoras Sensoriais/metabolismo , Células Receptoras Sensoriais/fisiopatologia , Papilas Gustativas/fisiopatologia , Distúrbios do Paladar/genética , Distúrbios do Paladar/fisiopatologia , Língua/fisiopatologia , Ubiquitina Tiolesterase/metabolismoRESUMO
ASIC3-immunoreactivity (ir) was examined in the rat vagal and glossopharyngeal sensory ganglia. In the jugular, petrosal and nodose ganglia, 24.8%, 30.8% and 20.6% of sensory neurons, respectively, were immunoreactive for ASIC3. These neurons were observed throughout the ganglia. A double immunofluorescence method demonstrated that many ASIC3-immunoreactive (ir) neurons co-expressed calcitonin gene-related peptide (CGRP)- or vanilloid receptor subtype 1 (VRL-1)-ir in the jugular (CGRP, 77.8%; VRL-1, 28.0%) and petrosal ganglia (CGRP, 61.7%; VRL-1, 21.5%). In the nodose ganglion, however, such neurons were relatively rare (CGRP, 6.3%; VRL-1, 0.4%). ASIC3-ir neurons were mostly devoid of tyrosine hydroxylase in these ganglia. However, some ASIC3-ir neurons co-expressed calbindin D-28k in the petrosal (5.5%) and nodose ganglia (3.8%). These findings may suggest that ASIC3-containing neurons have a wide variety of sensory modalities in the vagal and glossopharyngeal sensory ganglia.
Assuntos
Gânglios Sensitivos/citologia , Nervo Glossofaríngeo/citologia , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios Aferentes/metabolismo , Canais de Sódio/metabolismo , Nervo Vago/citologia , Canais Iônicos Sensíveis a Ácido , Animais , Calbindinas , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Contagem de Células/métodos , Imuno-Histoquímica/métodos , Ratos , Proteína G de Ligação ao Cálcio S100/metabolismo , Canais de Cátion TRPV/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Aspartate-immunoreactivity (ir) was examined in the mouse trigeminal ganglion (TG). The ir was detected in 34% of TG neurons and their cell bodies were of various sizes (mean +/- S.D. = 1,234 +/- 543 microm(2)). A triple immunofluorescence method revealed the co-expression of aspartate with calcitonin gene-related peptide (CGRP) and parvalbumin; 22% and 14% of aspartate-immunoreactive (ir) neurons were also immunoreactive for CGRP and parvalbumin, respectively. The co-expression of aspartate with both CGRP and parvalbumin was very rare in the TG. By retrograde tracing method, half and 66% of TG neurons which innervate the vibrissa and palate, respectively, contained aspartate-ir. The co-expression of aspartate with CGRP was more common among palatal neurons (36%) compared to vibrissal neurons (22%). Aspartate-ir neurons which co-expressed parvalbumin-ir were numerous in the vibrissa (17%) but not in the palate (4%). These findings may suggest that the function of aspartate-containing TG neurons is correlated with their peripheral receptive fields.
Assuntos
Ácido Aspártico/metabolismo , Neurônios Aferentes/metabolismo , Gânglio Trigeminal/citologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Contagem de Células/métodos , Tamanho Celular , Imuno-Histoquímica/métodos , Camundongos , Camundongos Endogâmicos C57BL , Parvalbuminas/metabolismo , Paladar/fisiologia , Vibrissas/inervação , Vibrissas/fisiologiaRESUMO
Immunohistochemistry for brain-derived neurotrophic factor (BDNF) was performed on the rat trigeminal ganglion (TG). The immunoreactivity (IR) was detected in 46% of TG neurons. These neurons were mostly small- or medium-sized (range, 149.7-1246.3 microm2; mean +/- SD = 373.4 +/- 151.6 microm2). A double immunofluorescence method also revealed that 54% of BDNF-immunoreactive (IR) neurons were immunoreactive for calcitonin-gene-related peptide. In addition, 93% of BDNF-IR TG neurons contained vanilloid receptor subtype 1. However, the co-expression of BDNF and vanilloid receptor 1-like receptor was very rare (less than 1%). In the trigeminal sensory nuclei, laminae II of the medullary dorsal horn was abundant in presumed BDNF-IR axon terminals. Such profiles were also detected in the dorsolateral part of the subnucleus oralis. The retrograde tracing and immunohistochemical methods demonstrated that BDNF-IR was common among cutaneous TG neurons (47%) but not tooth pulp TG neurons (13%). The present study indicates that BDNF-IR TG neurons have unmyelinated axons and project to the superficial medullary dorsal horn. It is likely that BDNF-containing neurons in both the trigeminal and spinal sensory systems have similarities in morphology and function. However, the content of BDNF in TG neurons probably depends on their peripheral targets. BDNF seems to convey nociceptive cutaneous input to the trigeminal sensory nuclei.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Neurônios Aferentes/metabolismo , Gânglio Trigeminal/citologia , Núcleos do Trigêmeo/citologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Contagem de Células/métodos , Tamanho Celular , Polpa Dentária/inervação , Polpa Dentária/fisiologia , Imuno-Histoquímica/métodos , Masculino , Ratos , Ratos Sprague-Dawley , Canais de Cátion TRPV/metabolismoRESUMO
The distribution of gamma and beta subunits of epithelial Na(+) channel (ENaC), markers for low-threshold mechanoreceptors in peripheral tissues, was examined in the tooth pulp. In the root pulp, gammaENaC- and betaENaC-immunoreactive (IR) nerve fibers showed a thick smooth appearance. These nerve fibers ascended toward the pulp horn and formed subodontoblastic nerve plexuses. Immunoelectron microscopic method revealed that 63% of axons were immunoreactive for gammaENaC in the root pulp. Virtually all myelinated axons showed gammaENaC-IR (97%), whereas unmyelinated axons were mostly devoid of it (12%). These findings suggest that myelinated tooth pulp nociceptors respond to mechanical stimuli.
Assuntos
Polpa Dentária/fisiologia , Dente Molar/fisiologia , Canais de Sódio/metabolismo , Animais , Polpa Dentária/inervação , Canais Epiteliais de Sódio , Imuno-Histoquímica , Masculino , Microscopia Imunoeletrônica , Dente Molar/inervação , Fibras Nervosas/fisiologia , Nociceptores/efeitos dos fármacos , Estimulação Física , Ratos , Ratos Sprague-DawleyRESUMO
The co-expression of calretinin with parvalbumin and calbindin D-28k was examined in the rat cranial and spinal sensory ganglia by triple immunofluorescence method. In the trigeminal and nodose ganglia, 9% and 5% of calretinin-immunoreactive neurons, respectively, also contained both parvalbumin- and calbindin D-28k immunoreactivity. These neurons had large cell bodies. In the trigeminal ganglion, they were restricted to the caudal portion. Such neurons were evenly distributed throughout the nodose ganglion. The co-expression could not be detected in the dorsal root, jugular or petrosal ganglia. Nerve fibers which co-expressed all the three calcium-binding proteins were observed in the inferior alveolar nerve but not the infraorbital nerve or palate. In the periodontal ligament, these nerve fibers formed Ruffini-like endings. These findings suggest that (1) the co-expression in trigeminal neurons is intimately related to their peripheral receptive fields; (2) the three calcium-binding proteins (calretinin, parvalbumin, calbindin D-28k) co-expressed in the trigeminal neurons may have mechanoreceptive function in the periodontal ligament.
Assuntos
Gânglios Sensitivos/metabolismo , Neurônios/fisiologia , Parvalbuminas/biossíntese , Proteína G de Ligação ao Cálcio S100/biossíntese , Proteína G de Ligação ao Cálcio S100/fisiologia , Medula Espinal/metabolismo , Animais , Calbindina 2 , Calbindinas , Técnica Indireta de Fluorescência para Anticorpo , Gânglios Espinais/citologia , Gânglios Espinais/metabolismo , Masculino , Mecanorreceptores/fisiologia , Fibras Nervosas/metabolismo , Gânglio Nodoso/citologia , Gânglio Nodoso/metabolismo , Ligamento Periodontal/inervação , Ligamento Periodontal/fisiologia , Ratos , Ratos Sprague-Dawley , Gânglio Trigeminal/citologia , Gânglio Trigeminal/metabolismoRESUMO
Previous studies indicate that descending modulation of nociception is progressively increased following persistent inflammation. The present study was designed to further examine the role of supraspinal neurons in descending modulation following persistent inflammation. Constant levels of paw withdrawal (PW) and tail flick (TF) latencies to noxious heat stimuli were achieved in lightly anesthetized rats (pentobarbital sodium 3-10 mg/kg/h, i.v.). Electrical stimulation (ES, 0.1 ms, 100 Hz, 20-200 A) was delivered to the rostral ventromedial medulla (RVM), mainly the nucleus raphe magnus (NRM). ES produced intensity-dependent inhibition of PW and TF. Following a unilateral hindpaw inflammation produced by injection of complete Freund's adjuvant (CFA), ES-produced inhibition underwent time-dependent changes. There was an initial decrease at 3 h after inflammation and a subsequent increase after inflammation in the excitability of RVM neurons and the inhibition of nocifensive responses. These changes were most robust after stimulation of the inflamed paw although similar findings were seen on the non-inflamed paw and tail. The inflammation-induced dynamic changes in descending modulation appeared to be correlated with changes in the activation of the N-methyl--aspartate (NMDA) excitatory amino acid receptor. Microinjection of an NMDA receptor antagonist, AP5 (1 pmol), resulted in an increase in the current intensity required for inhibition of the PW and TF. The effect of AP5 was less at 3 h after inflammation and significantly greater at 11-24 h after inflammation. In a subsequent experiment, ES-produced inhibition of nocifensive responses after inflammation was examined following selective chemical lesions of the nuclei reticularis gigantocellularis (NGC). Compared to vehicle-injected animals, microinjection of a soma-selective excitotoxin, ibotenic acid, enhanced ES-produced inhibition at 3 h but not at 24 h after inflammation. We propose that these time course changes reflect dynamic alterations in concomitant descending facilitation and inhibition. At early time points, NMDA receptor and NGC activation enhance descending facilitation; as time progresses, the dose-response curve of NMDA shifts to the left and descending inhibition dominates and masks any descending facilitation.
Assuntos
Hiperalgesia/fisiopatologia , Bulbo/fisiologia , Inibição Neural/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologia , 2-Amino-5-fosfonovalerato/farmacologia , Anestesia , Animais , Estimulação Elétrica , Agonistas de Aminoácidos Excitatórios , Antagonistas de Aminoácidos Excitatórios/farmacologia , Temperatura Alta , Ácido Ibotênico , Masculino , Inflamação Neurogênica/fisiopatologia , Nociceptores/fisiologia , Limiar da Dor/efeitos dos fármacos , Limiar da Dor/fisiologia , Ratos , Ratos Sprague-Dawley , Tempo de ReaçãoRESUMO
Treatment of pain in newborns is associated with problematic drug side effects. Previous studies demonstrate that an intraoral infusion of sucrose and other sweet components of mother's milk are effective in alleviating pain in infant rats and humans. These findings are of considerable significance, as sweet tastants are used in pain and stress management in a number of clinical procedures performed in human infants. The ability of sweet stimuli to induce analgesia is absent in adult rats, suggesting that this is a developmentally transient phenomenon. However, the age range over which intraoral sucrose is capable of producing analgesia is not known. We investigated the effects of intraoral sucrose (7.5%) on nocifensive withdrawal responses to thermal and mechanical stimuli in naive and inflamed rats at postnatal days (P) P0-21. In some rats, Complete Freund's adjuvant (CFA) was injected in a fore- or hindpaw to produce inflammation. In non-inflamed animals, for noxious thermal stimuli, sucrose-induced analgesia emerged at P3, peaked at P7-10, then progressively declined and was absent at P17. For mechanical forepaw stimuli, sucrose-induced analgesia emerged, and was maximal at approximately P10, then declined and was absent at P17. By contrast, maximal sucrose-induced analgesia for mechanical hindpaw stimuli was delayed (P13) compared to that for the forepaw, although it was also absent at P17. In inflamed animals, sucrose reduced hyperesthesia and hyperalgesia assessed with mechanical stimuli. Sucrose-induced analgesia in inflamed animals was initially present at P3 for the forepaw and P13 for the hindpaw, and was absent by P17 for both limbs. Intraoral sucrose produced significantly greater effects on responses in fore- and hindpaws in inflamed rats than in naive rats indicating that it reduces hyperalgesia and allodynia beyond its effects on responses in naive animals. These findings support the hypothesis that sucrose has a selective influence on analgesic mechanisms and that an enhanced sucrose effect takes place in hyperalgesic, inflamed animals as compared to naive animals. Taken together, these results indicate that intraoral sucrose alleviates transient pain in response to thermal and mechanical stimuli, and also effectively reduces inflammatory hyperalgesia and allodynia. Sucrose-induced analgesia is age-dependent and limited to the pre-weaning period in rats. The age-dependency of sucrose-induced analgesia and its differential maturation for the fore- and hindpaw may be due to developmental changes in endogenous analgesic mechanisms and developmental modulation of the interaction between gustatory and pain modulatory pathways.
Assuntos
Analgesia/métodos , Dor/tratamento farmacológico , Sacarose/farmacologia , Paladar/fisiologia , Doença Aguda , Administração Oral , Fatores Etários , Animais , Animais Recém-Nascidos , Doença Crônica , Modelos Animais de Doenças , Temperatura Alta , Limiar da Dor/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores Opioides/fisiologiaRESUMO
Brain stem descending pathways modulate spinal nociceptive transmission. In a lightly anesthetized rat preparation, we present evidence that such descending modulation undergoes time-dependent changes following persistent hindpaw inflammation. There was an initial decrease and a subsequent increase in the excitability of neurons in the rostral ventromedial medulla (RVM) involving facilitation and inhibition. These changes were most robust after stimulation of the inflamed paw although similar findings were seen on the non-inflamed paw and tail. The enhanced descending modulation appeared to be mediated by changes in the activation of the NMDA excitatory amino acid receptor. These findings demonstrate the dynamic plasticity of the pain modulating pathways in response to persistent tissue injury.
Assuntos
Inflamação/fisiopatologia , Bulbo/fisiopatologia , Plasticidade Neuronal , Dor/fisiopatologia , Animais , Comportamento Animal , Vias Eferentes/fisiopatologia , Estimulação Elétrica , Agonistas de Aminoácidos Excitatórios/farmacologia , Temperatura Alta , Masculino , N-Metilaspartato/farmacologia , Inibição Neural , Nociceptores/fisiopatologia , Medição da Dor , Estimulação Física , Ratos , Ratos Sprague-Dawley , Tempo de Reação , Receptores de Glutamato/fisiologia , Estimulação Química , Transmissão Sináptica/efeitos dos fármacosRESUMO
Noxious stimulus-evoked c-Fos expression in the spinal dorsal horn is modulated by noxious stimuli applied previously to remote areas of the body. To confirm the existence of such modulation in c-Fos expression in the trigeminal system, changes in c-Fos expression in the trigeminal nucleus caudalis induced by formalin injection into the rat whisker pad were examined by previously injecting formalin into different areas (contralateral whisker pad, ipsilateral or contralateral forepaw) of the body. Formalin injection-evoked c-Fos expression in this nucleus was significantly reduced by previous formalin injection into the contralateral whisker pad or ipsilateral forepaw but not into the contralateral forepaw. The interval between the two injections of formalin that produced a maximal reduction of formalin injection-evoked c-Fos expression was 1 h, and the reduction of c-Fos expression was less when the interval of the two noxious stimuli was longer or shorter than 1 h. These results suggested that noxious stimulus-evoked c-Fos expression in the trigeminal nucleus caudalis is reduced by noxious stimulus applied previously to remote areas, and the reduction is dependent on the area of previous noxious stimulation and interval between the two noxious stimuli.
Assuntos
Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Núcleo Inferior Caudal do Nervo Trigêmeo/metabolismo , Animais , Fixadores/farmacologia , Membro Anterior/efeitos dos fármacos , Membro Anterior/metabolismo , Formaldeído/farmacologia , Masculino , Neurônios/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Estimulação Química , Núcleo Inferior Caudal do Nervo Trigêmeo/efeitos dos fármacos , Vibrissas/efeitos dos fármacos , Vibrissas/metabolismoRESUMO
To determine the effects of nerve injury on Fos expression, temporal and spatial distributions of Fos-positive neurons in the trigeminal nucleus caudalis were examined after tissue injury for isolation of the infraorbital nerve as controls and transection of this nerve as well as noxious chemical stimulation by formalin injection in adult rats. Fos immunoreactivity was markedly elevated in laminae I and II of the only ipsilateral nucleus caudalis 2 h after these surgical procedures and noxious chemical stimulation. The distributions of Fos-positive neurons were restricted rostro-caudally following formalin injection and tissue injury compared to transection of the infraorbital nerve. One day after tissue injury and nerve transection, however, Fos-positive neurons were distributed bilaterally in laminae III and IV extending rostro-caudally and medio-laterally in this nucleus, and this persisted over the 2-week study period. The number of Fos-positive neurons in the side ipsilateral to nerve transection was markedly less than that in the contralateral side whereas positive neurons in the tissue injured rats were distributed symmetrically along the rostro-caudal axis. There was no difference in the contralateral sides between nerve transection and tissue injury groups. The rostro-caudal level showing reduction in Fos expression corresponded roughly to the sites of central termination of the injured nerve in this nucleus, suggesting a role for the primary afferents in the reduction of Fos expression in laminae III and IV neurons of the ipsilateral nucleus caudalis.
Assuntos
Proteínas do Tecido Nervoso/análise , Órbita/inervação , Proteínas Proto-Oncogênicas c-fos/análise , Núcleo Inferior Caudal do Nervo Trigêmeo/química , Animais , Imuno-Histoquímica , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
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.
Assuntos
Proteínas do Tecido Nervoso/metabolismo , Peptídeos/metabolismo , Gânglio Cervical Superior/metabolismo , Animais , Tamanho Celular , Colchicina/farmacologia , Face , Imuno-Histoquímica , Masculino , Músculo Masseter/inervação , NADPH Desidrogenase/metabolismo , Neurônios/metabolismo , Neuropeptídeo Y/metabolismo , Ratos , Ratos Sprague-Dawley , Pele/inervação , Glândula Submandibular/inervaçãoRESUMO
PURPOSE: To clarify the relationship between allodynia and the sprouting of myelinated fibers, we examined whether the administration of nerve growth factor (NGF) affected the paw withdrawal response to non-noxious mechanical stimuli and the sprouting of myelinated fibers into lamina II of the spinal dorsal horn, using a chronic constriction injury model of the sciatic nerve. METHODS: Mechanical allodynia was determined as the threshold of the withdrawal response stimulated by von Frey filaments. Sprouting was examined using horseradish peroxidase conjugated to the B fragment of cholera toxin (B-HRP). NGF was continuously infused into the site of nerve injury for 14 days after nerve ligation. RESULTS: With vehicle infusion, significantly increased responsiveness to mechanical stimuli was observed on postoperative days (PODs) 5, 7, and 14 after ligation, compared with before surgery, and B-HRP-positive fibers were newly localized in lamina II on PODs 7 and 14. Infusion of NGF reduced the responsiveness to mechanical stimuli on 5, 7, and 14 PODs and B-HRP-positive fibers in lamina II on PODs 7 and 14. CONCLUSION: We propose that the suppression of the increased responsiveness to mechanical stimuli produced by NGF could be related to the disappearance of B-HRP-positive fibers in lamina II.
RESUMO
Recent studies indicate that descending pain modulatory pathways undergo time-dependent changes in excitability following inflammation involving both facilitation and inhibition. The cellular and molecular mechanisms of these phenomena are unclear. In the present study, we examined N-methyl-D-aspartate (NMDA) receptor gene expression and neuronal activity in the rostral ventromedial medulla (RVM), a pivotal structure in pain modulatory circuitry, after complete Freund's adjuvant (CFA)-induced hindpaw inflammation. The reverse transcription polymerase chain reaction analysis indicated that there was an upregulation of mRNAs encoding NMDA receptor subunits in the RVM after inflammation. The increase in the NR1, NR2A, and NR2B receptor mRNAs started at 5 h, maintained for 1-7 days (P < 0.05-0.001) and returned to the control level at 14 days after inflammation. Western blot analysis indicated that the protein translation products of the NR2A subunit were also increased (P < 0.01). In single-unit extracellular recordings, we correlated RVM neuronal activity with the paw withdrawal response in rats with inflammation. We describe these RVM cells as on-, off-, and neutral-like cells because of their similarity to previous studies in which neuronal responses were correlated with tail-flick nocifensive behavior in the absence of inflammation. In contrast to previous studies in the absence of inflammation, using tail flick as a behavioral correlate, fewer off-like cells in naïve animals exhibited a complete pause before the paw withdrawal to a noxious thermal stimulus. The percentage of cells showing a pause of activity after noxious stimulation was further reduced after inflammation (chi(2) P < 0.0001 vs. naïve rats). Continuous neuronal recordings (3-6.5 h) revealed a phenotypic switch of RVM neurons during the development of inflammation: 11/15 neutral-like cells initially unresponsive to noxious stimuli exhibited and maintained response profiles characteristic of pain modulatory neurons (became off-like: n = 5; became on-like: n = 6). Neutral-like cells recorded in noninflamed animals did not show response profile changes during continuous recordings (5-5.5 h, n = 7). A population study (n = 165) confirmed an increase in on- and off-like cells and a decrease in neutral-like cells at 24 h after inflammation as compared with naïve rats (P < 0.001). These results suggest that enhanced NMDA receptor activation mediates time-dependent changes in excitability of RVM pain modulatory circuitry. The functional phenotypic switch of RVM neurons provides a novel mechanism underlying activity-dependent plasticity and enhanced net descending inhibition after inflammation.