RESUMO
AIM: Genetic factors are important for outcome after traumatic brain injury (TBI), although exact knowledge of relevant genes/pathways is still lacking. We here used an unbiased approach to define differentially activated pathways between the inbred DA and PVG rat strains. The results prompted us to study further if a naturally occurring genetic variation in glutathione-S-transferase alpha 4 (Gsta4) affects the outcome after TBI. RESULTS: Survival of neurons after experimental TBI is increased in PVG compared to the DA strain. Global expression profiling analysis shows the glutathione metabolism pathway to be the most regulated between the strains, with increased Gsta4 in PVG among top regulated transcripts. A congenic strain (R5) with a PVG genomic insert containing the Gsta4 gene on DA background displays a reversal of the strain pattern for Gsta4 expression and increased survival of neurons compared to DA. Gsta4 is known to effectively reduce 4-hydroxynonenal (4-HNE), a noxious by-product of lipid peroxidation. Immunostaining of 4-HNE was evident in both rat and human TBI. Intracerebral injection of 4-HNE resulted in neurodegeneration with increased levels of a marker for nerve injury in cerebrospinal fluid of DA compared to R5. INNOVATION: These findings provide strong support for the notion that the inherent capability of coping with increased 4-HNE after TBI affects outcome in terms of nerve cell loss. CONCLUSION: A naturally occurring variation in Gsta4 expression in rats affects neurodegeneration after TBI. Further studies are needed to explore if genetic variability in Gsta4 can be associated to outcome also in human TBI.
Assuntos
Lesões Encefálicas/enzimologia , Lesões Encefálicas/patologia , Glutationa Transferase/genética , Isoenzimas/genética , Neurônios/enzimologia , Neurônios/patologia , Aldeídos/metabolismo , Aldeídos/toxicidade , Animais , Lesões Encefálicas/genética , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Humanos , Isoenzimas/metabolismo , Peroxidação de Lipídeos , Masculino , Camundongos , Camundongos TransgênicosRESUMO
Increasing evidence suggests that genetic background affects outcome of traumatic brain injuries (TBI). Still, there is limited detailed knowledge on what pathways/processes are affected by genetic heterogeneity. The inbred rat strains DA and PVG differ in neuronal survival following TBI. We here carried out global expressional profiling to identify differentially regulated pathways governing the response to an experimental controlled brain contusion injury. One of the most differentially regulated molecular networks concerned immune cell trafficking. Subsequent characterization of the involved cells using flow cytometry demonstrated greater infiltration of neutrophils and monocytes, as well as a higher degree of microglia activation in DA compared to PVG rats. In addition, DA rats displayed a higher number of NK cells and a higher ratio of CD161bright compared to CD161dim NK cells. Local expression of complement pathway molecules such as C1 and C3 was higher in DA and both the key complement component C3 and membrane-attack complex (MAC) could be demonstrated on axons and nerve cells. A stronger activation of the complement system in DA was associated with higher cerebrospinal fluid levels of neurofilament-light, a biomarker for nerve/axonal injury. In summary, we demonstrate substantial differences between DA and PVG rats in activation of inflammatory pathways; in particular, immune cell influx and complement activation associated with neuronal/axonal injury after TBI. These findings suggest genetic influences acting on inflammatory activation to be of importance in TBI and motivate further efforts using experimental forward genetics to identify genes/pathways that affect outcome.
Assuntos
Lesões Encefálicas , Ativação do Complemento , Leucócitos , RNA Mensageiro/análise , Ratos Endogâmicos , Animais , Lesões Encefálicas/genética , Lesões Encefálicas/imunologia , Movimento Celular/genética , Ativação do Complemento/genética , Ativação do Complemento/imunologia , Complemento C1q/genética , Complemento C1q/imunologia , Complemento C3/genética , Complemento C3/imunologia , Complexo de Ataque à Membrana do Sistema Complemento/genética , Complexo de Ataque à Membrana do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/genética , Proteínas do Sistema Complemento/imunologia , Citocinas/genética , Citocinas/imunologia , Perfilação da Expressão Gênica , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Leucócitos/citologia , Leucócitos/imunologia , Microglia/citologia , Microglia/imunologia , Monócitos/citologia , Monócitos/imunologia , Subfamília B de Receptores Semelhantes a Lectina de Células NK/genética , Subfamília B de Receptores Semelhantes a Lectina de Células NK/imunologia , Neutrófilos/citologia , Neutrófilos/imunologia , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Ratos Endogâmicos/genética , Ratos Endogâmicos/imunologiaRESUMO
Genetic regulation of autoimmune neuroinflammation is a well known phenomenon, but genetic influences on inflammation following traumatic nerve injuries have received little attention. In this study we examined the inflammatory response in a rat traumatic brain injury (TBI) model, with a particular focus on major histocompatibility class II (MHC II) presentation, in two inbred rat strains that have been extensively characterized in experimental autoimmune encephalomyelitis (EAE); DA and PVG. In addition, MHC and Vra4 congenic strains on these backgrounds were studied to give information on MHC and non-MHC gene contribution. Thus, allelic differences in Vra4, harboring the Ciita gene, was found to regulate expression of the invariant chain at the mRNA level, with a much smaller effect exerted by the MHC locus itself. Notably, however, at the protein level the MHC congenic PVG-RT1(av1) strain displayed much stronger MHCII(+) presentation, as shown both by immunolabeling and flow cytometry, than the PVG strain, dwarfing the effect of Ciita. The PVG-RT1(av1) strain had significantly more T-cell influx than both DA and PVG, suggesting regulation both by MHC and non-MHC genes. Finally, in terms of outcome, the EAE susceptible DA strain displayed a significantly smaller resulting lesion volume than the resistant PVG-RT1(av1) strain. These results provide additional support for a role of adaptive immune response after neurotrauma and demonstrate that outcome is significantly affected by host genetic factors.
Assuntos
Lesões Encefálicas/imunologia , Inflamação/imunologia , Complexo Principal de Histocompatibilidade/imunologia , Linfócitos T/imunologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/imunologia , Encéfalo/patologia , Lesões Encefálicas/patologia , Citometria de Fluxo , Genes MHC da Classe II/efeitos dos fármacos , Genes MHC da Classe II/imunologia , Genes MHC da Classe II/fisiologia , Inflamação/fisiopatologia , Complexo Principal de Histocompatibilidade/efeitos dos fármacos , Complexo Principal de Histocompatibilidade/fisiologia , Masculino , Proteína Básica da Mielina/farmacologia , Fragmentos de Peptídeos/farmacologia , Ratos , Ratos Endogâmicos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T/fisiologiaRESUMO
Secondary brain damage following traumatic brain injury in part depends on neuroinflammation, a process where genetic factors may play an important role. We examined the response to a standardized cortical contusion in two different inbred rat strains, Dark Agouti (DA) and Piebald Virol Glaxo (PVG). Both are well characterized in models of autoimmune neuroinflammation, where DA is susceptible and PVG resistant. We found that infiltration of polymorphonuclear granulocytes (PMN) at 3-day postinjury was more pronounced in PVG. DA was more infiltrated by T cells at 3-day postinjury, showed an enhanced glial activation at 7-day postinjury and higher expression of C3 complement at 7-day postinjury. Neurodegeneration, assessed by Fluoro-Jade, was also more pronounced in the DA strain at 30-day postinjury. These results demonstrate differences in the response to cortical contusion injury attributable to genetic influences and suggest a link between injury-induced inflammation and neurodegeneration. Genetic factors that regulate inflammation elicited by brain trauma may be important for the development of secondary brain damage.
Assuntos
Lesões Encefálicas , Proteínas do Sistema Complemento/metabolismo , Regulação da Expressão Gênica/fisiologia , Microglia/metabolismo , Degeneração Neural/etiologia , Animais , Lesões Encefálicas/complicações , Lesões Encefálicas/metabolismo , Lesões Encefálicas/patologia , Antígeno CD11b/metabolismo , Proteínas do Sistema Complemento/genética , Modelos Animais de Doenças , Ectodisplasinas/metabolismo , Fluoresceínas , Leucossialina/metabolismo , Masculino , Degeneração Neural/patologia , Neutrófilos/metabolismo , Compostos Orgânicos , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Estatísticas não Paramétricas , Linfócitos T/metabolismoRESUMO
BACKGROUND: Damage to nerve cells and axons leading to neurodegeneration is a characteristic feature of many neurological diseases. The degree of genetic influence on susceptibility to axotomy-induced neuronal death has so far been unknown. We have examined two gene regions, Vra1 and Vra2, previously linked to nerve cell loss after ventral root avulsion in a rat F2 intercross between the DA and PVG inbred rat strains. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we use two generations (G8 and G10 cohorts) of an advanced intercross line between DA and PVG(av1) to reproduce linkage to Vra1 and to fine-map this region. By isolating the effect from Vra1 in congenic strains, we demonstrate that Vra1 significantly regulates the loss of motoneurons after avulsion. The regulatory effect mediated by Vra1 thus resides in a congenic fragment of 9 megabases. Furthermore, we have used the advanced intercross lines to give more support to Vra2, originally detected as a suggestive QTL. CONCLUSIONS/SIGNIFICANCE: The results demonstrated here show that naturally occurring allelic variations affect susceptibility to axotomy-induced nerve cell death. Vra1 and Vra2 represent the first quantitative trait loci regulating this phenotype that are characterized and fine mapped in an advanced intercross line. In addition, congenic strains provide experimental evidence for the Vra1 effect on the extent of injury-induced neurodegeneration. Identification of the underlying genetic variations will increase our understanding of the regulation and mechanisms of neurodegeneration.
Assuntos
Mapeamento Cromossômico/métodos , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Alelos , Animais , Sobrevivência Celular , Cruzamentos Genéticos , Feminino , Ligação Genética , Variação Genética , Genótipo , Haplótipos , Humanos , Masculino , Ratos , Medula Espinal/patologiaRESUMO
Patients who have suffered nerve injury show profound inter-individual variability in neuropathic pain even when the precipitating injury is nearly identical. Variability in pain behavior is also observed across inbred strains of mice where it has been attributed to genetic polymorphisms. Identification of cellular correlates of pain variability across strains can advance the understanding of underlying pain mechanisms. Voltage-gated sodium channels (VGSCs) play a major role in the generation and propagation of action potentials in the primary afferents and are therefore of obvious importance for pain phenotype. Here, we examined the mRNA expression levels of the VGSC alpha-subunits Na(v)1.3, Na(v)1.5, Na(v)1.6, and Na(v)1.7, as well as the auxiliary VGSC-related molecule, Contactin. Dorsal root ganglia (DRG) and spinal cords from 5 inbred mouse strains with contrasting pain phenotype (AKR/J, C3H/HeJ, C57BL/6J, C58/J and CBA/J) were analyzed 7 days following sciatic and saphenous nerve transection. In the DRG, Na(v)1.6, Na(v)1.7 and Contactin were abundantly expressed in control animals. Following nerve injury, the residual mRNA levels of Na(v)1.6 (downregulated in two of the strains) correlated tightly to the extent of autotomy behavior. A suggestive correlation was also seen for the post-injury mRNA levels of Contactin (downregulated in all strains) with autotomy. Thus, our results suggest a contribution by DRG Na(v)1.6, and possibly Contactin to neuropathic pain in the neuroma model in mice.
Assuntos
Gânglios Espinais/metabolismo , Doenças do Sistema Nervoso Periférico/metabolismo , Automutilação/metabolismo , Células Receptoras Sensoriais/metabolismo , Canais de Sódio/metabolismo , Medula Espinal/metabolismo , Animais , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Contactinas , Modelos Animais de Doenças , Gânglios Espinais/fisiopatologia , Regulação da Expressão Gênica/genética , Ativação do Canal Iônico/genética , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Canal de Sódio Disparado por Voltagem NAV1.6 , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Neuralgia/genética , Neuralgia/metabolismo , Neuralgia/fisiopatologia , Nociceptores/metabolismo , Doenças do Sistema Nervoso Periférico/genética , Doenças do Sistema Nervoso Periférico/fisiopatologia , Fenótipo , Células do Corno Posterior/metabolismo , RNA Mensageiro/metabolismo , Neuropatia Ciática/genética , Neuropatia Ciática/metabolismo , Neuropatia Ciática/fisiopatologia , Automutilação/genética , Automutilação/fisiopatologia , Canais de Sódio/genética , Especificidade da Espécie , Medula Espinal/fisiopatologiaRESUMO
Major histocompatibility complex (MHC) class II is of critical importance for the induction of immune responses. Levels of MHC class II in the nervous system are normally low, but expression is up-regulated in many disease conditions. In rat and human, variation in the MHC class II transactivator gene (C2ta) is associated with differential expression of MHC class II and susceptibility to autoimmune disease. Here we have characterized the response to facial nerve transection in 7 inbred mouse strains (C57BL/6J, DBA/2J, 129X1/SvJ, BALB/cJ, SJL/J, CBA/J, and NOD). The results demonstrate differences in expression of C2ta and markers for MHC class I and II expression, glial activation, and T cell infiltration. Expression levels of C2ta and Cd74 followed similar patterns, in contrast to MHC class I and markers of glial activation. The regulatory region of the C2ta gene was subsequently sequenced in the four strains (C57BL/6/J, DBA/2J, SJL/J and 129X1/SvJ) that represented the phenotypical extremes with regard to C2ta/Cd74 expression. We found 3 single nucleotide polymorphisms in the type I (pI) and type III (pIII) promoters of C2ta, respectively. Higher expression of pI in 129X1/SvJ correlated with the pI haplotype specific for this strain. Furthermore, congenic strains carrying the 129X1/SvJ C2ta allele on B6 background displayed significantly higher C2ta and Cd74 expression compared to parental controls. We conclude that genetic polymorphisms in the type I promoter of C2ta regulates differential expression of MHC class II, but not MHC class I, Cd3 and other markers of glial activation.
Assuntos
Traumatismos do Nervo Facial/imunologia , Genes MHC da Classe II , Proteínas Nucleares/genética , Regiões Promotoras Genéticas , Transativadores/genética , Animais , Antígenos de Diferenciação de Linfócitos B/análise , Antígenos de Diferenciação de Linfócitos B/genética , Complexo CD3/análise , Feminino , Regulação da Expressão Gênica , Variação Genética , Antígenos de Histocompatibilidade Classe II/análise , Antígenos de Histocompatibilidade Classe II/genética , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos , Proteínas Nucleares/análise , Polimorfismo Genético , Isoformas de Proteínas , Especificidade da Espécie , Transativadores/análiseRESUMO
Local CNS inflammation takes place in many neurological disorders and is important for autoimmune neuroinflammation. Microglial activation is strain-dependent in rats and differential MHC class II expression is influenced by variations in the Mhc2ta gene. Despite sharing Mhc2ta and MHC class II alleles, BN and LEW.1N rats differ in MHC class II expression after ventral root avulsion (VRA). We studied MHC class II expression and glial activation markers in BN rats after VRA. Our results demonstrate that MHC class II expression originates from a subpopulation of IBA1(+), ED1(-), and ED2(-) microglia. We subsequently performed a genome-wide linkage scan in an F2(BNxLEW.1N) population, to investigate gene regions regulating this inflammatory response. Alongside MHC class II, we studied the expression of MHC class I, co-stimulatory molecules, complement components, microglial markers and Il1b. MHC class II and other transcripts were commonly regulated by gene regions on chromosomes 1 and 7. Furthermore, a common region on chromosome 10 regulated expression of complement and co-stimulatory molecules, while a region on chromosome 11 regulated MHC class I. We also detected epistatic interactions in the regulation of the inflammatory process. These results reveal the complex regulation of CNS inflammation by several gene regions, which may have relevance for disease.
Assuntos
Encefalite/imunologia , Genes MHC da Classe II , Microglia/fisiologia , Locos de Características Quantitativas , Animais , Antígenos de Diferenciação de Linfócitos B/genética , Mapeamento Cromossômico , Epistasia Genética , Feminino , Regulação da Expressão Gênica , Ligação Genética , Antígenos de Histocompatibilidade Classe II/genética , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos LewRESUMO
We have recently shown that the major histocompatibility complex (MHC) exerts a regulatory influence on the development of neuropathic pain-like behaviors after partial sciatic nerve injury in male rats. In the present study, we assessed the role of the MHC in peripheral nerve injury-induced pain as well as central pain following spinal cord injury in female rats using the following inbred strains: Dark Agouti (DA; RT1(av1)), Piebald Virol Glaxo (PVG; RT1(c)) and in the MHC-congenic strain PVG-RT1(av1). In line with our previous observation in male rats, PVG-RT1(c) displayed more severe allodynia compared to the strains carrying the RT1(av1) haplotype (PVG-RT1(av1) and DA-RT1(av1)) following sciatic nerve injury in female rats. However, the MHC did not seem to impact the development of allodynia following spinal cord injury since the two congenic strains PVG-RT1(c) and PVG-RT1(av) did not differ after spinal cord injury. Interestingly, the DA-RT1(av1) strain displayed significantly more severe allodynia than both PVG strains and this difference was not explained by the extent of spinal cord injury. These results suggest that there are differences in the genetic mechanisms for neuropathic pain development following peripheral or central nervous system injury, both in regarding to the role of the MHC complex as well as non-MHC genes.
Assuntos
Complexo Principal de Histocompatibilidade/genética , Neuralgia/genética , Neuropatia Ciática/genética , Isquemia do Cordão Espinal/genética , Animais , Animais Congênicos , Feminino , Hiperalgesia/genética , Masculino , Ratos , Neuropatia Ciática/complicações , Isquemia do Cordão Espinal/complicaçõesRESUMO
We have previously demonstrated that differences in neuropathic pain-like behaviors after sciatic nerve injury genetically maps to the major histocompatibility complex (MHC) in rats carrying RT1(c) or RT1(av1) haplotypes on the Piebald Virol Glaxo (PVG) background. In order to further explore the genetic contribution to neuropathic pain, we have here examined the MHC-congenic rat strains PVG-RT1(n) and PVG-RT1(av1) and the inbred strains PVG (RT1(c)) and Brown-Norway (BN; RT1(n)). All studied strains developed mechanical hypersensitivity (allodynia-like behavior) of the hind paw after photochemically induced sciatic nerve injury. However, the PVG-RT1(n) and PVG strains displayed significantly more allodynia than PVG-RT1(av1) and BN rats. In addition, the BN strain demonstrated an elevated threshold for the baseline response. The results demonstrate that both MHC and non-MHC genes influence experimental neuropathic pain in rats and also suggest that allelic variation contained in the RT1(av1) haplotype on the PVG background protects against neuropathic pain.
Assuntos
Complexo Principal de Histocompatibilidade/genética , Neuralgia/genética , Animais , Animais Congênicos , Ratos , Nervo Isquiático/lesõesRESUMO
Herpes simplex encephalitis (HSE) is characterized by severe focal brain inflammation leading to substantial loss of nervous tissue. The authors established a model of Herpes simplex virus type 1 (HSV)-1-induced acute encephalitis in the rat by injecting into the whiskers' area a virus strain isolated from a fatal human HSE case. The model might resemble natural propagation of HSV-1 in humans; spreading from the mouth and lips via the trigeminal nerve to trigeminal ganglia and subsequently entering the central nervous system (CNS). HSV-1 infected Dark Agouti (DA) rats developed a well-synchronized disease and died 5 days after inoculation. HSV-1 detection by quantitative polymerase chain reaction (qPCR), virus isolation and immunohistochemistry, magnetic resonance imaging, and histopathological examination verified dramatic encephalitis mainly in the brainstem, but also in the olfactory bulb and other segments of the brain of diseased rats. In contrast, Piebald Virol Glaxo (PVG) rats were completely resistant to disease, displaying a more rapid clearance of peripheral infection and no evidence of virus entering into neither the trigeminal ganglia nor the CNS. These results suggest a regulation of susceptibility to HSV-1-induced encephalitis at the level of peripheral infection and subsequent neuronal uptake/transport of the virus. This provides a basis for future positioning of genetic polymorphisms regulating HSE and for dissection of important pathogenetic mechanisms of this severe human disease.
Assuntos
Encefalite por Herpes Simples/imunologia , Encefalite por Herpes Simples/microbiologia , Herpesvirus Humano 1/isolamento & purificação , Animais , Tronco Encefálico/metabolismo , DNA Viral/análise , Modelos Animais de Doenças , Suscetibilidade a Doenças , Encefalite por Herpes Simples/genética , Encefalite por Herpes Simples/patologia , Humanos , Ratos , Gânglio Trigeminal/microbiologiaRESUMO
Presentation of Ag bound to MHC class II (MHC II) molecules to CD4+ T cells is a key event in adaptive immune responses. Genetic differences in MHC II expression in the rat CNS were recently positioned to allelic variability in the CIITA gene (Mhc2ta), located within the Vra4 locus on rat chromosome 10. In this study, we have examined reciprocal Vra4-congenic strains on the DA and PVGav1 backgrounds, respectively. After experimental nerve injury the strain-specific MHC II expression on microglia was reversed in the congenic strains. Similar findings were obtained after intraparenchymal injection of IFN-gamma in the brain. Expression of MHC class II was also lower on B cells and dendritic cells from the DA.PVGav1-Vra4- congenic strain compared with DA rats after in vitro stimulation with IFN-gamma. We next explored whether Vra4 may affect the outcome of experimental autoimmune disease. In experimental autoimmune encephalomyelitis induced by immunization with myelin oligodendrocyte glycoprotein, DA.PVGav1-Vra4 rats displayed a lower disease incidence and milder disease course compared with DA, whereas both PVGav1 and PVGav1.DA-Vra4 rats were completely protected. These results demonstrate that naturally occurring allelic differences in Mhc2ta have profound effects on the quantity of MHC II expression in the CNS and on immune cells and that this genetic variability also modulates susceptibility to autoimmune neuroinflammation.
Assuntos
Alelos , Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/imunologia , Predisposição Genética para Doença , Antígenos de Histocompatibilidade Classe II/genética , Proteínas Nucleares/genética , Transativadores/genética , Animais , Animais Congênicos , Células Cultivadas , Regulação para Baixo/genética , Regulação para Baixo/imunologia , Encefalomielite Autoimune Experimental/patologia , Feminino , Marcadores Genéticos , Antígenos de Histocompatibilidade Classe II/biossíntese , Masculino , Inflamação Neurogênica/genética , Inflamação Neurogênica/imunologia , Proteínas Nucleares/biossíntese , Ratos , Ratos Endogâmicos , Rizotomia , Raízes Nervosas Espinhais/patologia , Raízes Nervosas Espinhais/cirurgia , Transativadores/biossínteseRESUMO
Neuropathic pain is a common consequence of damage to the nervous system. We here report a genetic analysis of development of neuropathic pain-like behaviors after unilateral photochemically-induced ischemic sciatic nerve injury in a panel of inbred rat strains known to display different susceptibility to autoimmune neuroinflammation. Pain behavior was initially characterized in Dark-Agouti (DA; RT1(av1)), Piebald Virol Glaxo (PVG; RT1(c)), and in the major histocompatibility complex (MHC)-congenic strain PVG-RT1(av1). All strains developed mechanical hypersensitivity (allodynia) following nerve injury. However, the extent and duration of allodynia varied significantly among the strains, with PVG displaying more severe allodynia compared to DA rats. Interestingly, the response of PVG-RT1(avRT1) was similar to that of DA, suggesting regulation by the MHC locus. This notion was subsequently confirmed in an F2 cohort derived from crossing of the PVG and PVG-RT1(av1)strains, where allodynia was reduced in homozygous or heterozygous carriers of the RT1(av1) allele in comparison to rats homozygous for the RT1(c) allele. These results indicate that certain allelic variants of the MHC could influence susceptibility to develop and maintain neuropathic pain-like behavior following peripheral nerve injury in rats.
Assuntos
Comportamento Animal , Hiperestesia/genética , Complexo Principal de Histocompatibilidade/genética , Neuralgia/genética , Doenças do Sistema Nervoso Periférico/genética , Tato/genética , Animais , Feminino , Predisposição Genética para Doença/genética , Masculino , Ratos , Especificidade da EspécieRESUMO
Inflammation is a common characteristic of spinal cord injury. The nature of this response, whether it is beneficial or detrimental, has been the subject of debate. It has been reported that susceptibility to autoimmunity is correlated with increased functional impairment following spinal cord injury. As the ability to mount an autoimmune response has most consistently been associated with certain haplotypes of the major histocompatibility complex (MHC), we analysed the possible effects of the MHC haplotype on functional impairment and recovery following spinal cord injury. A contusion injury was induced in experimental autoimmune encephalomyelitis-susceptible and -resistant rats [Dark Agouti, Lewis and Piebald Viral Glaxo (PVG), respectively]. We found that locomotion recovered significantly better in Dark Agouti rats compared with PVG and Lewis rats but an F2 intercross (PVG x PVG-RT1(av1)) excluded the possibility that this difference was MHC haplotype-dependent. Thus, we conclude that recovery following spinal cord injury is subject to considerable genetic heterogeneity that is not coupled to the MHC haplotype region. Continued research of genetic variants regulating recovery following spinal cord injury is warranted.
Assuntos
Complexo Principal de Histocompatibilidade , Ratos Endogâmicos/genética , Recuperação de Função Fisiológica/genética , Traumatismos da Medula Espinal/genética , Traumatismos da Medula Espinal/fisiopatologia , Análise de Variância , Animais , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Comportamento Animal , Ectodisplasinas/metabolismo , Feminino , Ligação Genética , Antígenos de Histocompatibilidade Classe I/metabolismo , Locomoção/genética , Imageamento por Ressonância Magnética , Masculino , Ratos , Índice de Gravidade de Doença , Especificidade da Espécie , Fatores de TempoRESUMO
Numerous CNS diseases of primarily non-inflammatory origin, such as idiopathic neurodegenerative diseases, contain elements of inflammation, with T cell infiltration, MHC class II expression and neuron/axon damage. Gene mapping in human clinical materials have in most cases failed to unravel discrete genes, since most genes instrumental in non-Mendelian forms of these complex diseases are likely to modestly affect risk, be evolutionary conserved in the population and vary between individuals. We here describe the exploration of susceptibility to neurodegeneration and inflammatory glial activation in response to mechanical nerve injury using experimental genetic models. The response to ventral root avulsion, which is a simple and reproducible model of nerve injury-induced neurodegeneration and inflammation, was examined in a panel of inbred rat strains. A whole genome scan subsequently performed in a F2(DAxPVG) intercross identified quantitative trait loci (QTLs) regulating different features of the nerve injury response. Fine mapping in an advanced intercross line revealed polymorphisms in the Mhc2ta gene as being responsible for strain differences in MHC class II expression. Furthermore, a polymorphism in the syntenic human gene, MHC2TA, was associated both with lower expression of MHC class II-associated genes and increased susceptibility to inflammatory diseases. These results provide important insights into the genetic regulation of fundamental physiological responses of the nervous system to damage and demonstrate relevance also for human diseases.
Assuntos
Genes MHC da Classe II/genética , Doenças Neurodegenerativas/genética , Proteínas Nucleares/genética , Locos de Características Quantitativas/genética , Transativadores/genética , Animais , Axônios/imunologia , Cruzamentos Genéticos , Modelos Animais de Doenças , Predisposição Genética para Doença , Masculino , Doenças Neurodegenerativas/imunologia , Proteínas Nucleares/imunologia , Polimorfismo Genético , Locos de Características Quantitativas/imunologia , Ratos , Ratos Endogâmicos , Especificidade da Espécie , Raízes Nervosas Espinhais/imunologia , Raízes Nervosas Espinhais/lesões , Transativadores/imunologiaRESUMO
Axonal damage, a core feature of neurological diseases, induces a retrograde reaction in neurons and surrounding glia. We determined transcriptional profiles of this reaction using Affymetrix oligonucleotide arrays. Gene expression was examined in spinal cord tissue prior to injury and following ventral root avulsion in two inbred rat strains, where the degree of neurodegeneration differs. Stringent statistical analysis revealed 278 regulated genes, whereof 245 were regulated by the injury and 68 differed between strains. Principal component analysis disclosed a common injury response pattern significantly modified by genetic background. Notably, inflammatory genes comprised the largest group of genes induced by injury and these transcripts prevailed in the strain most susceptible to neurodegeneration. In addition, levels of the strain regulated genes C1qb and Timp1 correlated with degree of neurodegeneration in a cohort of genetically heterogeneous animals. These results suggest a link between the inflammatory response elicited by nerve injury and subsequent neurodegeneration.
Assuntos
Inflamação/genética , Degeneração Neural/genética , Animais , Axotomia , Morte Celular/genética , Proliferação de Células , Sobrevivência Celular/genética , Sobrevivência Celular/fisiologia , Citoesqueleto/metabolismo , DNA Complementar/biossíntese , DNA Complementar/genética , Interpretação Estatística de Dados , Matriz Extracelular/metabolismo , Expressão Gênica/fisiologia , Regulação da Expressão Gênica/fisiologia , Receptores de Hialuronatos/genética , Imuno-Histoquímica , Hibridização In Situ , Lisossomos/metabolismo , Masculino , Proteínas Mitocondriais , Neurônios/metabolismo , Neurônios/ultraestrutura , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo/fisiologia , Análise de Componente Principal , Ratos , Ratos Endogâmicos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transmissão Sináptica/fisiologia , Inibidor Tecidual de Metaloproteinase-1/genéticaRESUMO
Inflammation and neurodegeneration characterize multiple sclerosis, as well as many other diseases of the central nervous system (CNS). The understanding of the molecular pathways that regulate these processes is of fundamental importance for the development of new therapies. Nerve lesions paradigms in animals can serve as important tools to dissect central features of human CNS disease and by using these models certain key regulators have also been identified. However, our knowledge of how aspects of neurodegeneration and CNS inflammation are regulated on a genomic level is very limited. Such knowledge may help to unravel disease mechanisms. By using a standardized nerve trauma model, ventral root avulsion (VRA), in a series of inbred rat strains we here demonstrate a potent genetic regulation of the degree of neuron death and glial activation. Genome wide mapping of these phenotypes in experimental rat strain crosses identifies several quantitative trait loci (QTLs) controlling nerve lesion-induced nerve cell death, local T cell accumulation and expression of MHC class II on microglia. This approach may lead to the identification of evolutionary conserved genetic polymorphisms in key controlling genes, which can serve as prime candidates for association studies in several human CNS diseases.
Assuntos
Doenças do Sistema Nervoso Central/genética , Mapeamento Cromossômico/métodos , Doenças Neurodegenerativas/genética , Animais , Doenças do Sistema Nervoso Central/patologia , Cruzamentos Genéticos , Modelos Animais de Doenças , Humanos , Inflamação/genética , Modelos Biológicos , Doenças Neurodegenerativas/patologia , Neuroglia/fisiologiaRESUMO
Expression of T cell immunoglobulin- and mucin-domain-containing molecules (TIMs) can be used as T helper (Th) differentiation markers in the human and mouse. We examined the expression of TIM-1 and -3 mRNAs in rat MBP(63-88)-induced experimental autoimmune encephalomyelitis (EAE). TIM-3 expression was upregulated in the spinal cord during EAE and following antigen restimulation of the encephalitogenic TCRBV8S2+ population. Interestingly, TIM-3 expression was also detected by in situ hybridization in resident cells of the nervous system. TIM-1 was expressed in B cells but not in resident CNS cells and TIM-1 mRNA levels in spinal cord were unchanged throughout the course of EAE. These results support the notion that TIM-3 can also be used as a Th1 differentiation marker in the rat. However, expression of TIM-1 and -3 is not restricted solely to T cells and the presence of TIM-3 in resident CNS cells may indicate a role for this molecule in the interaction between the nervous and immune systems.
Assuntos
Encefalomielite Autoimune Experimental/metabolismo , Regulação da Expressão Gênica/fisiologia , Sistema Imunitário/metabolismo , Proteínas de Membrana/metabolismo , Sistema Nervoso/metabolismo , Receptores Virais/metabolismo , Animais , Células Cultivadas , Encefalomielite Autoimune Experimental/induzido quimicamente , Feminino , Citometria de Fluxo/métodos , Receptor Celular 1 do Vírus da Hepatite A , Receptor Celular 2 do Vírus da Hepatite A , Hibridização In Situ/métodos , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Proteínas de Membrana/genética , Proteína Básica da Mielina , Sistema Nervoso/patologia , Fragmentos de Peptídeos , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos Lew , Receptores Virais/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
Antigen presentation to T cells by MHC molecules is essential for adaptive immune responses. To determine the exact position of a gene affecting expression of MHC molecules, we finely mapped a previously defined rat quantitative trait locus regulating MHC class II on microglia in an advanced intercross line. We identified a small interval including the gene MHC class II transactivator (Mhc2ta) and, using a map over six inbred strains combined with gene sequencing and expression analysis, two conserved Mhc2ta haplotypes segregating with MHC class II levels. In humans, a -168A --> G polymorphism in the type III promoter of the MHC class II transactivator (MHC2TA) was associated with increased susceptibility to rheumatoid arthritis, multiple sclerosis and myocardial infarction, as well as lower expression of MHC2TA after stimulation of leukocytes with interferon-gamma. We conclude that polymorphisms in Mhc2ta and MHC2TA result in differential MHC molecule expression and are associated with susceptibility to common complex diseases with inflammatory components.
Assuntos
Artrite Reumatoide/genética , Complexo Principal de Histocompatibilidade , Esclerose Múltipla/genética , Infarto do Miocárdio/genética , Proteínas Nucleares/genética , Transativadores/genética , Animais , Predisposição Genética para Doença , Genótipo , Antígenos de Histocompatibilidade Classe II/imunologia , Polimorfismo de Nucleotídeo Único , Ratos , Medula Espinal/imunologiaRESUMO
Recently, MHC class I molecules have been shown to be important for the retraction of synaptic connections that normally occurs during development [Huh, G.S., Boulanger, L. M., Du, H., Riquelme, P. A., Brotz, T. M. & Shatz, C. J. (2000) Science 290, 2155-2158]. In the adult CNS, a classical response of neurons to axon lesion is the detachment of synapses from the cell body and dendrites. We have investigated whether MHC I molecules are involved also in this type of synaptic detachment by studying the synaptic input to sciatic motoneurons at 1 week after peripheral nerve transection in beta2-microglobulin or transporter associated with antigen processing 1-null mutant mice, in which cell surface MHC I expression is impaired. Surprisingly, lesioned motoneurons in mutant mice showed more extensive synaptic detachments than those in wild-type animals. This surplus removal of synapses was entirely directed toward inhibitory synapses assembled in clusters. In parallel, a significantly smaller population of motoneurons reinnervated the distal stump of the transected sciatic nerve in mutants. MHC I molecules, which traditionally have been linked with immunological mechanisms, are thus crucial for a selective maintenance of synapses during the synaptic removal process in neurons after lesion, and the lack of MHC I expression may impede the ability of neurons to regenerate axons.