RESUMEN
Schwann cells (SCs) critically maintain the plasticity of the peripheral nervous system. Peripheral nerve injuries and infections stimulate SCs in order to retrieve homeostasis in neural tissues. Previous studies indicate that Mycobacterium leprae (ML) regulates the expression of key factors related to SC identity, suggesting that alterations in cell phenotype may be involved in the pathogenesis of neural damage in leprosy. To better understand whether ML restricts the plasticity of peripheral nerves, the present study sought to determine the expression of Krox-20, Sox-10, c-Jun and p75NTR in SC culture and mice sciatic nerves, both infected by ML Thai-53 strain. Primary SC cultures were stimulated with two different multiplicities of infection (MOI 100:1; MOI 50:1) and assessed after 7 and 14 days. Sciatic nerves of nude mice (NU-Foxn1nu ) infected with ML were evaluated after 6 and 9 months. In vitro results demonstrate downregulation of Krox-20 and Sox-10 along with the increase in p75NTR-immunolabelled cells. Concurrently, sciatic nerves of infected mice showed a significant decrease in Krox-20 and increase in p75NTR. Our results corroborate previous findings on the interference of ML in the expression of factors involved in cell maturation, favouring the maintenance of a non-myelinating phenotype in SCs, with possible implications for the repair of adult peripheral nerves.
Asunto(s)
Regulación hacia Abajo , Proteína 2 de la Respuesta de Crecimiento Precoz/biosíntesis , Lepra/metabolismo , Células de Schwann/metabolismo , Nervio Ciático/metabolismo , Animales , Diferenciación Celular/fisiología , Células Cultivadas , Modelos Animales de Enfermedad , Lepra/microbiología , Lepra/patología , Ratones Desnudos , Mycobacterium leprae/aislamiento & purificación , Plasticidad Neuronal/fisiología , Receptores de Factor de Crecimiento Nervioso/metabolismo , Células de Schwann/microbiología , Células de Schwann/patología , Nervio Ciático/microbiología , Nervio Ciático/patología , Técnicas de Cultivo de TejidosRESUMEN
Inflammation in Guillain-Barré syndrome (GBS) is manifested by changes in matrix metalloproteinase (MMP) and pro-inflammatory cytokine expression. We investigated the expression of MMP-2, -9 and TNF-α and correlated it with pathological changes in sciatic nerve tissue from Campylobacter jejuni-induced chicken model for GBS. Campylobacter jejuni and placebo were fed to chickens and assessed for disease symptoms. Sciatic nerves were examined by histopathology and immunohistochemistry. Expressions of MMPs and TNF-α, were determined by real-time PCR, and activities of MMPs by zymography. Diarrhea developed in 73.3% chickens after infection and 60.0% of them developed GBS like neuropathy. Pathology in sciatic nerves showed perinodal and/or patchy demyelination, perivascular focal lymphocytic infiltration and myelin swelling on 10th- 20th post infection day (PID). MMP-2, -9 and TNF-α were up-regulated in progressive phase of the disease. Enhanced MMP-2, -9 and TNF-α production in progressive phase correlated with sciatic nerve pathology in C. jejuni-induced GBS chicken model.
Asunto(s)
Infecciones por Campylobacter/enzimología , Campylobacter jejuni/fisiología , Síndrome de Guillain-Barré/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Parálisis/enzimología , Animales , Infecciones por Campylobacter/genética , Infecciones por Campylobacter/microbiología , Infecciones por Campylobacter/patología , Campylobacter jejuni/genética , Pollos , Modelos Animales de Enfermedad , Síndrome de Guillain-Barré/genética , Síndrome de Guillain-Barré/microbiología , Síndrome de Guillain-Barré/patología , Humanos , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Parálisis/genética , Parálisis/microbiología , Nervio Ciático/enzimología , Nervio Ciático/microbiología , Nervio Ciático/patología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Demyelination is a common pathologic feature in many neurodegenerative diseases including infection with leprosy-causing Mycobacterium leprae. Because of the long incubation time and highly complex disease pathogenesis, the management of nerve damage in leprosy, as in other demyelinating diseases, is extremely difficult. Therefore, an important challenge in therapeutic interventions is to identify the molecular events that occur in the early phase before the progression of the disease. Here we provide evidence that M. leprae-induced demyelination is a result of direct bacterial ligation to and activation of ErbB2 receptor tyrosine kinase (RTK) signaling without ErbB2-ErbB3 heterodimerization, a previously unknown mechanism that bypasses the neuregulin-ErbB3-mediated ErbB2 phosphorylation. MEK-dependent Erk1 and Erk2 (hereafter referred to as Erk1/2) signaling is identified as a downstream target of M. leprae-induced ErbB2 activation that mediates demyelination. Herceptin (trastuzumab), a therapeutic humanized ErbB2-specific antibody, inhibits M. leprae binding to and activation of ErbB2 and Erk1/2 in human primary Schwann cells, and the blockade of ErbB2 activity by the small molecule dual ErbB1-ErbB2 kinase inhibitor PKI-166 (ref. 11) effectively abrogates M. leprae-induced myelin damage in in vitro and in vivo models. These results may have implications for the design of ErbB2 RTK-based therapies for both leprosy nerve damage and other demyelinating neurodegenerative diseases.
Asunto(s)
Enfermedades Desmielinizantes/metabolismo , Lepra/metabolismo , Mycobacterium leprae/metabolismo , Receptor ErbB-2/metabolismo , Transducción de Señal , Animales , Anticuerpos Monoclonales/farmacología , Anticuerpos Monoclonales Humanizados , Butadienos/farmacología , Células COS , Células Cultivadas , Chlorocebus aethiops , Técnicas de Cocultivo , Enfermedades Desmielinizantes/patología , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Células HeLa , Humanos , Lepra/microbiología , Ratones , Ratones Noqueados , Ratones Desnudos , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Mycobacterium leprae/genética , Nitrilos/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Ratas , Células de Schwann/enzimología , Células de Schwann/metabolismo , Nervio Ciático/metabolismo , Nervio Ciático/microbiología , Nervio Ciático/ultraestructura , TrastuzumabRESUMEN
Neuroactive steroids act in the peripheral nervous system as physiological regulators and as protective agents for acquired or inherited peripheral neuropathy. In recent years, modulation of neuroactive steroids levels has been studied as a potential therapeutic approach to protect peripheral nerves from damage induced by diabetes. Nuclear receptors of the liver X receptor (LXR) family regulate adrenal steroidogenesis via their ability to control cholesterol homeostasis. Here we show that rat sciatic nerve expresses both LRXα and ß isoforms and that these receptors are functional. Activation of liver X receptors using a synthetic ligand results in increased levels of neurosteroids and protection of the sciatic nerve from neuropathy induced by diabetes. LXR ligand treatment of streptozotocin-treated rats increases expression in the sciatic nerve of steroidogenic acute regulatory protein (a molecule involved in the transfer of cholesterol into mitochondria), of the enzyme P450scc (responsible for conversion of cholesterol into pregnenolone), of 5α-reductase (an enzyme involved in the generation of neuroactive steroids) and of classical LXR targets involved in cholesterol efflux, such as ABCA1 and ABCG1. These effects were associated with increased levels of neuroactive steroids (e.g., pregnenolone, progesterone, dihydroprogesterone and 3α-diol) in the sciatic nerve, and with neuroprotective effects on thermal nociceptive activity, nerve conduction velocity, and Na(+), K(+)-ATPase activity. These results suggest that LXR activation may represent a new pharmacological avenue to increase local neuroactive steroid levels that exert neuroprotective effects in diabetic neuropathy.
Asunto(s)
Neuropatías Diabéticas/metabolismo , Neuropatías Diabéticas/prevención & control , Receptores Nucleares Huérfanos/metabolismo , Esteroides/metabolismo , Esteroides/uso terapéutico , Análisis de Varianza , Animales , Benzoatos/farmacología , Bencilaminas/farmacología , Peso Corporal/efectos de los fármacos , Cromatografía Liquida/métodos , Diabetes Mellitus Experimental/complicaciones , Neuropatías Diabéticas/fisiopatología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Hiperalgesia/etiología , Ligandos , Receptores X del Hígado , Masculino , Proteínas de la Mielina/genética , Proteínas de la Mielina/metabolismo , Conducción Nerviosa/fisiología , Umbral del Dolor , Ratas , Ratas Sprague-Dawley , Nervio Ciático/metabolismo , Nervio Ciático/microbiología , Nervio Ciático/patología , Nervio Ciático/fisiopatología , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Espectrometría de Masas en Tándem/métodosAsunto(s)
Absceso/microbiología , Bloqueo Nervioso/efectos adversos , Nervio Ciático/microbiología , Infecciones Estafilocócicas/microbiología , Muslo/microbiología , Absceso/cirugía , Anestesiología , Femenino , Humanos , Persona de Mediana Edad , Bloqueo Nervioso/instrumentación , Infecciones Estafilocócicas/cirugía , Staphylococcus aureus/aislamiento & purificación , Muslo/inervación , Muslo/cirugíaRESUMEN
INTRODUCTION: We employed a nanosilver-collagen scaffold and tested its effects on inhibiting bacteria and facilitating nerve regeneration. METHODS: Based on our previous research, we prepared bionic scaffolds with different concentrations of nanosilver and examined their internal structures by scanning electron microscopy and energy dispersive spectroscopy. We implanted these scaffolds or autologous nerve grafts into rats to repair a 10-mm injury of the sciatic nerve. RESULTS: The 2 mg/ml group showed a >10 mm bacterial inhibition zone in all 3 types of bacterial culture dishes. At day 60 postsurgery, the 2 mg/ml group also showed the highest amplitude of evoked potential (AMP) and nerve conduction velocity (NCV). The regenerating nerves in the 2 mg/ml group were denser and more mature, and with thicker and well-arrayed myelin sheath. CONCLUSIONS: These results demonstrate that nanosilver scaffolds (2 mg/ml group) were effective in inhibiting bacteria both in vitro and in vivo, and reduced the contamination-caused immune responses, which in turn promoted nerve regeneration and functional recovery.
Asunto(s)
Nanopartículas , Nervio Ciático/microbiología , Nervio Ciático/cirugía , Plata/farmacología , Ingeniería de Tejidos , Andamios del Tejido , Animales , Escherichia coli/efectos de los fármacos , Recuento de Leucocitos , Microscopía Electrónica de Transmisión , Modelos Animales , Músculo Esquelético/patología , Regeneración Nerviosa , Tamaño de los Órganos , Pseudomonas aeruginosa/efectos de los fármacos , Ratas Sprague-Dawley , Nervio Ciático/lesiones , Nervio Ciático/ultraestructura , Staphylococcus aureus/efectos de los fármacosRESUMEN
Colchicine was used to inhibit axonal transport and to demonstrate that rabies virus spread from the peripheral inoculation site to the CNS by the retrograde axoplasmic flow. Colchicine was applied by the mean of elastomer implants around the sciatic nerve of young rats in order to obtain higher local concentrations of the drug. This procedure avoided the systemic effects of colchicine encountered with the usual treatment. To confirm the efficiency of the axoplasmic flow inhibition by colchicine, 125I-tetanus toxin was used as a marker. Uptake of colchicine by the sciatic nerve was monitored by the use of 3H-labelled colchicine. Interruption of the retrograde axoplasmic flow resulted in prevention of fixed and street rabies virus propagation. Moreover, the centrifugal spread of rabies could be inhibited using this experimental procedure.
Asunto(s)
Transporte Axonal , Axones/microbiología , Virus de la Rabia , Animales , Anticuerpos Antivirales/análisis , Antígenos Virales/análisis , Transporte Axonal/efectos de los fármacos , Axones/inmunología , Colchicina/metabolismo , Colchicina/farmacología , Femenino , Virus de la Rabia/efectos de los fármacos , Virus de la Rabia/inmunología , Ratas , Nervio Ciático/inmunología , Nervio Ciático/metabolismo , Nervio Ciático/microbiologíaRESUMEN
Borna disease (BD) is an immune-mediated neurological disease caused by infection of the nervous system with a negative strand RNA virus, Borna disease virus (BDV). The host range for BDV is broad and extends from birds to primates. A BDV-like agent may cause disease in humans. Until recently, BDV-infected neural cells could only be identified immunocytochemically using serum from BDV-infected animals. The advent of BDV cDNA clones allowed definition of the relationship between viral nucleic acids and viral proteins in vivo. In situ hybridization with strand-specific RNA probes from a BDV cDNA clone, pAF4, identified BDV genomic RNA and BDV mRNAs in neurons, astrocytes, Schwann cells and ependymal cells in an anatomic distribution consistent with that of BDV proteins. Genomic RNA was contained primarily within the nucleus, whereas mRNAs were found in both the nuclear and cytoplasmic compartments. Viral RNAs were demonstrated in neurons expressing BDV proteins and in glial cells by combined techniques of immunocytochemistry and in situ hybridization.
Asunto(s)
Astrocitos/microbiología , Enfermedad de Borna/patología , Virus de la Enfermedad de Borna/aislamiento & purificación , Encéfalo/microbiología , Epéndimo/microbiología , Células de Schwann/microbiología , Nervio Ciático/microbiología , Animales , Animales Recién Nacidos , Astrocitos/patología , Virus de la Enfermedad de Borna/genética , Encéfalo/patología , ADN Viral/análisis , ADN Viral/genética , Epéndimo/patología , Genes Virales , Técnicas para Inmunoenzimas , Inmunohistoquímica , Masculino , Hibridación de Ácido Nucleico , ARN Mensajero/análisis , ARN Mensajero/genética , Ratas , Ratas Endogámicas Lew , Células de Schwann/patología , Nervio Ciático/patología , Proteínas Virales/análisisRESUMEN
Rhombencephalitis due to Listeria monocytogenes is characterized by progressive cranial nerve palsies and subacute inflammation in the brain stem. In this paper, we report observations made on mice infected with L. monocytogenes. Unilateral inoculation of bacteria into facial muscle, or peripheral parts of a cranial nerve, induced clinical and histological signs of mainly ipsilateral rhombencephalitis. Similarly, unilateral inoculation of bacteria into lower leg muscle or peripheral parts of sciatic nerve was followed by lumbar myelitis. In these animals, intraaxonal bacteria were seen in the sciatic nerve and its corresponding nerve roots ipsilateral to the bacterial application site. Development of myelitis was prevented by transsection of the sciatic nerve proximally to the hindleg inoculation site. Altogether, our results support the hypothesis that Listeria rhombencephalitis is caused by intraaxonal bacterial spread from peripheral sites to the central nervous system.
Asunto(s)
Axones/microbiología , Sistema Nervioso Central/microbiología , Listeria monocytogenes/patogenicidad , Meningitis por Listeria/fisiopatología , Nervios Periféricos/microbiología , Animales , Transporte Axonal/fisiología , Axones/metabolismo , Axones/patología , Tronco Encefálico/microbiología , Tronco Encefálico/patología , Tronco Encefálico/fisiopatología , Sistema Nervioso Central/patología , Sistema Nervioso Central/fisiopatología , Nervio Facial/microbiología , Nervio Facial/patología , Nervio Facial/fisiopatología , Femenino , Meningitis por Listeria/patología , Ratones , Ratones Endogámicos ICR , Nervios Periféricos/patología , Nervios Periféricos/fisiopatología , Nervio Ciático/microbiología , Nervio Ciático/patología , Nervio Ciático/fisiopatología , Médula Espinal/microbiología , Médula Espinal/patología , Médula Espinal/fisiopatologíaRESUMEN
This report describes a mammalian model for exploring the role of virus in peripheral neuropathy. Schwann cells in culture were permissive for mouse cytomegalovirus (MCMV) replication. Intraperitoneal inoculation rarely led to sciatic nerve infection. Sciatic nerves infected by direct intraneural injection produced infectious virus and contained viral antigen at 4 days postinfection (pi). Nerves taken later, at 4 to 8 weeks pi, contained no infectious virus, but MCMV was present in a latent state because culture of nerve explants reactivated virus. The findings contrast the viral permissiveness of cultured Schwann cells to the latency observed in intact peripheral nerve.
Asunto(s)
Citomegalovirus/crecimiento & desarrollo , Nervio Ciático/microbiología , Animales , Citomegalovirus/genética , Citomegalovirus/aislamiento & purificación , Replicación del ADN , Ratones , Técnicas de Cultivo de Órganos , Células de Schwann/microbiología , Replicación ViralRESUMEN
Herpes simplex type 1 virus was inoculated into 3-week-old mice via four different routes; intracerebral, intravenous, intranasal and directly into the sciatic nerve. Virus antigen-containing cells in the central nervous system were identified by both an immunofluorescence and immunoperoxidase method. The portal of entry of virus into the CNS appeared to be the major determinant of distribution of virus antigen. Direct haematogenous seeding of virus into the CNS was not proven. It seems probable that infection was first established in sensory ganglia. Within the CNS, regions of high virus antigen concentration paralleled high cell density suggesting cell to cell spread. Consistent involvement of certain neuron groups may be due to their selective vulnerability. These animal experiments provide some explanation for the patterns of CNS herpetic infection observed in man.
Asunto(s)
Antígenos Virales/análisis , Sistema Nervioso Central/microbiología , Encefalitis/transmisión , Infecciones por Herpesviridae/transmisión , Animales , Encéfalo/microbiología , Ratones , Ratones Endogámicos BALB C , Cavidad Nasal/microbiología , Vías Olfatorias/microbiología , Nervio Ciático/microbiología , Médula Espinal/microbiología , Viremia/microbiologíaRESUMEN
Previous studies of peripherally injected mouse scrapie suggested that invasion of the CNS occurs initially in mid-thoracic cord by neural spread of infection from spleen and other visceral sites of extraneural replication. We now show that infection of the left sciatic nerve leads to direct spread of infection to brain (at a rate of approximately 1.0-2.0 mm/day), bypassing the need for extraneural replication and thus producing shorter incubation periods. However, the efficiency of intraneural infection is low. It can be increased by crush injury or by the injection of lysophosphatidyl choline, both of which temporarily increase the surface area of axolemma exposed to inoculum. Once infection is established, agent seems to spread throughout the nervous system but, at the clinical stage of disease, the titres in the PNS are much lower than in the CNS.
Asunto(s)
Encéfalo/microbiología , Nervio Ciático/microbiología , Scrapie/microbiología , Médula Espinal/microbiología , Animales , Femenino , Ratones , Ratones Endogámicos , Priones/patogenicidad , Ovinos , Bazo/microbiología , Replicación ViralRESUMEN
Four-week-old Wistar rats were inoculated with HEV by different routes. Animals died of encephalitis after intraperitoneal (i.p.), subcutaneous (s.c.) and intravenous (i.v.) as well as intracerebral (i.c.) and intranasal (i.n.) inoculation. However when inoculated subcutaneously, rats died a few days earlier than those inoculated i.p. and i.v., suggesting that the virus might be transmitted to the central nervous system (CNS) by the neuronal route rather than by blood stream. Rats which were inoculated subcutaneously at the site of the neck (group A) began to die on day 4 p.i., a few days earlier than animals inoculated in the foot pad of the right leg (group B). On day 2 and 3 after inoculation, the virus titer in the brain was higher in group A, but group B animals showed higher virus titers in the lumber region of spinal cord than group A animals. In order to follow the virus spread from the peripheral nerve to the brain, the virus was inoculated into the sciatic nerve of rats. The inoculated rats developed clinical signs on day 4 and began to die on day 6. On day 2, virus was detected in the posterior half of the spinal cord and migrated toward the anterior half and in the brain where it was present on day 3. The highest virus titers in the brain were recorded on day 4 to 6, meanwhile the virus titers in the spinal cord tend to decrease. By immunohistochemical study, antigen positive neurons were found in the spinal cord and brain on day 4.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Corteza Cerebral/microbiología , Coronavirus/fisiología , Encefalomielitis/microbiología , Vías Nerviosas/microbiología , Médula Espinal/microbiología , Administración Intranasal , Animales , Antígenos Virales/análisis , Encéfalo , Coronavirus/aislamiento & purificación , Inyecciones , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Inyecciones Subcutáneas , Neuronas/microbiología , Ratas , Ratas Wistar , Nervio Ciático/microbiología , Factores de TiempoRESUMEN
Herpes simplex virus type 1 (HSV-1) replicated productively in rabbit and guinea pig ganglia and nerve organ cultures when inoculated in high titres. Treatment with IgG 20 hr before and 48 hr after infection produced a delay of 4 to 7 days in the recovery of HSV-1 by the method of co-cultivation. The same result was obtained when IgG was combined with human leukocyte interferon. There was no difference in the period up to HSV recovery between the groups treated with interferon alone and the HSV control. Morphological evidence by light and electron microscopy of viral productive infection was obtained in all the cell types of nervous tissues infected in vitro.
Asunto(s)
Ganglios Espinales/microbiología , Inmunoglobulina G/inmunología , Interferones/farmacología , Nervio Ciático/microbiología , Simplexvirus/efectos de los fármacos , Replicación Viral/efectos de los fármacos , Animales , Cápside/ultraestructura , Núcleo Celular/microbiología , Citoplasma/microbiología , Cobayas , Humanos , Leucocitos , Técnicas de Cultivo de Órganos , Conejos , Simplexvirus/crecimiento & desarrollo , Simplexvirus/ultraestructuraRESUMEN
Swiss albino mice were inoculated with Mycobacterium leprae obtained from untreated lepromatous patients. Histopathological study of sciatic nerves showed no abnormality. However a few free acid fast bacilli (AFB) were detected in the sciatic nerves taken from the inoculated limbs during the early stages of infection, suggesting the nerve-fibre route of travel as seen in humans in experimental leprosy, too.
Asunto(s)
Lepra/patología , Nervio Ciático/patología , Animales , Humanos , Ratones , Ratones Endogámicos , Mycobacterium leprae/aislamiento & purificación , Nervio Ciático/microbiologíaRESUMEN
Ultrastructural observation of sciatic nerves from eight Armadillos were made. Six animals had intravenous inoculation of M. leprae, one had of foot pad, while one had natural leprosy. The available nerves were biopsied at various time sequence ranging from five weeks to twenty four months. Semithin sections did not reveal any neuropathy. Ultrastructurally perineurium was thick and endoneurial collagen was increased. Initially demyelination of non-myelinated fibres was seen in all nerves irrespective of mode of infection. This was followed by demyelination of small myelinated fibres. Active remyelination was predominantly after 17 months. Schwann cell activity was increased and various stages of division were seen. Bacilli were extracellular, intraxonal, in endothelium and in perineurium. Significant observations were on blood vessels. These observations are discussed.
Asunto(s)
Lepra/patología , Nervio Ciático/ultraestructura , Animales , Armadillos , Membrana Basal/inmunología , Vasos Sanguíneos/ultraestructura , Femenino , Lepra/microbiología , Masculino , Microscopía Electrónica , Células de Schwann/ultraestructura , Nervio Ciático/irrigación sanguínea , Nervio Ciático/microbiologíaRESUMEN
Mouse sciatic nerves were subjected to devascularization, M. leprae inoculation, and combined insult of devascularization + footpad inoculation (FPI). Changes were seen in FPI nerves only after eight months, but in cases of combined insult, changes were evident in hours. Both the groups showed initial loss of small myelinated fibres. No proliferation of Schwann cells was in FPI nerves, but in combined insult it was maximum after two weeks. Presence of M. leprae seems to be arresting Schwann cell activity after two weeks. Blood vessels showed increased endothelial cell cytoplasm, basement membrane proliferation and villi formation. These changes seem to be specific of endoneurial blood vessels of leprosy nerves. Increased number of mast cells seems to be specific of devascularized and FPI nerves. Increased number of macrophages expressed low immunity of devascularized nerves. Eosinophils migrated to endoneurium as a result of leakage of axoplasm.