RESUMO
BACKGROUND: Group B Streptococcus (GBS) and Streptococcus pneumoniae (SP) are leading causes of bacterial meningitis in neonates and children. Each pathogen produces a pore-forming cytolytic toxin, ß-hemolysin/cytolysin (ß-h/c) by GBS and pneumolysin by SP. The aim of this study was to understand the role of these pore-forming cytotoxins, in particular of the GBS ß-h/c, as potential neurotoxins in experimental neonatal meningitis. METHODS: Meningitis was induced in 7- and 11-day-old rats by intracisternal injection of wild type (WT) GBS or SP and compared with isogenic ß-h/c- or pneumolysin-deficient mutants, or a double mutant of SP deficient in pneumolysin and hydrogen peroxide production. RESULTS: GBS ß-h/c and SP pneumolysin contributed to neuronal damage, worsened clinical outcome and weight loss, but had no influence on the early kinetics of leukocyte influx and bacterial growth in the cerebrospinal fluid. In vitro, ß-h/c-induced neuronal apoptosis occurred independently of caspase-activation and was not preventable by the broad spectrum caspase-inhibitor z-VAD-fmk. CONCLUSIONS: These data suggest that both cytolytic toxins, the GBS ß-h/c and SP pneumolysin, contribute to neuronal damage in meningitis and extend the concept of a key role for bacterial pore-forming cytolysins in the pathogenesis and sequelae of neonatal meningitis.
Assuntos
Citotoxinas/toxicidade , Meningite/microbiologia , Meningite/patologia , Proteínas Citotóxicas Formadoras de Poros/toxicidade , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/patologia , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Encéfalo/citologia , Caspases/metabolismo , Células Cultivadas , Citotoxinas/metabolismo , Embrião de Mamíferos , Neurônios/efeitos dos fármacos , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Ratos , Ratos Wistar , Streptococcus agalactiae/metabolismo , Streptococcus pneumoniae/metabolismoRESUMO
Major barriers separating the blood from tissue compartments in the body are composed of endothelial cells. Interaction of bacteria with such barriers defines the course of invasive infections, and meningitis has served as a model system to study endothelial cell injury. Here we report the impressive ability of Streptococcus pneumoniae, clinically one of the most important pathogens, to induce 2 morphologically distinct forms of programmed cell death (PCD) in brain-derived endothelial cells. Pneumococci and the major cytotoxins H2O2 and pneumolysin induce apoptosis-like PCD independent of TLR2 and TLR4. On the other hand, pneumococcal cell wall, a major proinflammatory component, causes caspase-driven classical apoptosis that is mediated through TLR2. These findings broaden the scope of bacterial-induced PCD, link these effects to innate immune TLRs, and provide insight into the acute and persistent phases of damage during meningitis.
Assuntos
Apoptose , Cerebelo/fisiopatologia , Células Endoteliais/metabolismo , Meningite Pneumocócica/fisiopatologia , Streptococcus pneumoniae , Animais , Apoptose/efeitos dos fármacos , Proteínas de Bactérias/farmacologia , Caspases/metabolismo , Parede Celular/metabolismo , Células Cultivadas , Cerebelo/citologia , Cerebelo/patologia , Células Endoteliais/microbiologia , Células Endoteliais/patologia , Peróxido de Hidrogênio/farmacologia , Glicoproteínas de Membrana/deficiência , Glicoproteínas de Membrana/metabolismo , Meningite Pneumocócica/microbiologia , Meningite Pneumocócica/patologia , Camundongos , Camundongos Knockout , Ratos , Ratos Wistar , Receptores de Superfície Celular/deficiência , Receptores de Superfície Celular/metabolismo , Estreptolisinas/farmacologia , Receptor 2 Toll-Like , Receptor 4 Toll-Like , Receptores Toll-LikeRESUMO
Stroke affects the normally well-balanced interplay of the 2 supersystems: the nervous and the immune system. Recent research elucidated some of the involved signals and mechanisms and, importantly, was able to demonstrate that brain-immune interactions are highly relevant for functional outcome after stroke. Immunodepression after stroke increases the susceptibility to infection, the most relevant complication in stroke patients. However, immunodepression after stroke may also have beneficial effects, for example, by suppressing autoaggressive responses during lesion-induced exposure of central nervous system-specific antigens to the immune system. Thus, before immunomodulatory therapy can be applied to stroke patients, we need to understand better the interaction of brain and immune system after focal cerebral ischemia. Until then, anticipating an important consequence of stroke-induced immunodepression, bacterial infection, preventive antibiotic strategies have been proposed. In mouse experiments, preventive antibiotic treatment dramatically improves mortality and outcome. Results of clinical studies on this issue are contradictory at present, and larger trials are needed to settle the question whether (and which) stroke patients should be preventively treated. Nevertheless, clinical evidence is emerging demonstrating that stroke-induced immunodepression in humans not only exists, but has very similar features to those characterized in rodent experiments.
Assuntos
Isquemia Encefálica/complicações , Síndromes de Imunodeficiência/etiologia , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/imunologia , Animais , Isquemia Encefálica/tratamento farmacológico , Isquemia Encefálica/imunologia , Humanos , Síndromes de Imunodeficiência/tratamento farmacológico , Acidente Vascular Cerebral/tratamento farmacológicoRESUMO
Pneumococcus is the most common and aggressive cause of bacterial meningitis and induces a novel apoptosis-inducing factor-dependent (AIF-dependent) form of brain cell apoptosis. Loss of production of two pneumococcal toxins, pneumolysin and H(2)O(2), eliminated mitochondrial damage and apoptosis. Purified pneumolysin or H(2)O(2) induced microglial and neuronal apoptosis in vitro. Both toxins induced increases of intracellular Ca(2+) and triggered the release of AIF from mitochondria. Chelating Ca(2+) effectively blocked AIF release and cell death. In experimental pneumococcal meningitis, pneumolysin colocalized with apoptotic neurons of the hippocampus, and infection with pneumococci unable to produce pneumolysin and H(2)O(2) significantly reduced damage. Two bacterial toxins, pneumolysin and, to a lesser extent, H(2)O(2), induce apoptosis by translocation of AIF, suggesting new neuroprotective strategies for pneumococcal meningitis.
Assuntos
Peróxido de Hidrogênio/toxicidade , Meningite Pneumocócica/etiologia , Estreptolisinas/toxicidade , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Fator de Indução de Apoptose , Proteínas de Bactérias , Sinalização do Cálcio/efeitos dos fármacos , Células Cultivadas , Flavoproteínas/biossíntese , Humanos , Proteínas de Membrana/biossíntese , Meningite Pneumocócica/microbiologia , Meningite Pneumocócica/patologia , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Microscopia Eletrônica , Modelos Neurológicos , Ratos , Streptococcus pneumoniae/patogenicidadeRESUMO
BACKGROUND AND PURPOSE: Bacterial pneumonia is the most common cause of death in patients sustaining acute stroke and is believed to result from an increased aspiration. Recently, stroke-induced immunodeficiency was described in a mouse model of cerebral ischemia, which is primarily caused by overactivation of sympathetic nervous system. We tested if stroke-induced immunodeficiency increases the risk of pneumonia after aspiration in a newly developed model of poststroke pneumonia. METHODS: Experimental stroke in mice was induced by occlusion of the middle cerebral artery (MCAO) for 60 minutes. Aspiration pneumonia was induced by intranasal application of 20 microL of a defined suspension of Streptococcus pneumoniae in phosphate-buffered saline 4 or 14 days after MCAO. Treatment comprised moxifloxacin (100 mg/kg body weight, six times every 2 hours after operation) or propranolol (30 mg/kg body weight, immediately before as well as 4 and 8 hours after MCAO). Readout was lung histology and bacterial counts in lung and blood. RESULTS: Nasal inoculation of only 200 colony-forming units of S. pneumoniae caused severe pneumonia and bacteremia after experimental stroke, whereas 200,000 colony-forming units are needed to induce comparable disease in sham animals. Aspiration pneumonia in stroke animals outlasted acute stroke state but was preventable by beta-adrenoreceptor blockade. CONCLUSIONS: Experimental stroke propagates bacterial aspiration from harmless intranasal colonization to harmful pneumonia. Prevention of infections by beta-adrenoreceptor blockade suggests that immunodepression by sympathetic hyperactivity is essential for progression of bacterial aspiration to pneumonia.
Assuntos
Síndromes de Imunodeficiência/etiologia , Infarto da Artéria Cerebral Média/complicações , Inalação , Pneumonia Bacteriana/etiologia , Acidente Vascular Cerebral/complicações , Doença Aguda , Administração Intranasal , Antagonistas Adrenérgicos beta/uso terapêutico , Animais , Bacteriemia/etiologia , Catecolaminas/fisiologia , Progressão da Doença , Suscetibilidade a Doenças , Feminino , Síndromes de Imunodeficiência/fisiopatologia , Síndromes de Imunodeficiência/prevenção & controle , Infarto da Artéria Cerebral Média/imunologia , Masculino , Camundongos , Cavidade Nasal/microbiologia , Pneumonia Pneumocócica/etiologia , Organismos Livres de Patógenos Específicos , Streptococcus pneumoniae , Acidente Vascular Cerebral/imunologia , Sistema Nervoso Simpático/efeitos dos fármacos , Sistema Nervoso Simpático/fisiopatologiaRESUMO
Voltage-gated calcium channels (VGCC) and N-methyl-d-aspartate receptors (NMDAR) account for most of the depolarization-induced neuronal calcium entry. The susceptibility of individual routes of calcium entry for nitric oxide (NO) is largely unknown. We loaded cultured rat cortical neurons with fluo-4 acetoxymethylester to study the effect of the NO synthase inhibitor Nomega-nitro-l-arginine and the NO donor S-nitroso-N-acetylpenicillamine on the intracellular calcium concentration ([Ca2+]i). The potassium-induced [Ca2+]i increase was amplified by Nomega-nitro-l-arginine and attenuated by S-nitroso-N-acetylpenicillamine. This modulation was abolished by either the P/Q-type VGCC antagonist omega-agatoxin IVA or by the NMDAR antagonist MK-801, but not by N-type (omega-conotoxin GVIA) or L-type (nimodipine) VGCC blockers. These results suggest that NO can modulate neuronal calcium entry during depolarization by interacting with P/Q-type VGCC and NMDAR.
Assuntos
Canais de Cálcio Tipo P/metabolismo , Canais de Cálcio Tipo Q/metabolismo , Cálcio/metabolismo , Neurônios/metabolismo , Óxido Nítrico/fisiologia , Receptores de N-Metil-D-Aspartato/metabolismo , Potenciais de Ação/fisiologia , Animais , Células Cultivadas , Líquido Extracelular/fisiologia , Líquido Intracelular/metabolismo , Neocórtex/citologia , Neocórtex/metabolismo , Neurônios/citologia , Ratos , Transdução de Sinais/fisiologiaRESUMO
We report a case of atypical cerebral venous drainage in a 38-year-old woman with symptoms of benign paroxysmal positional vertigo. Thrombosis of the left internal jugular vein and sigmoid sinus was suspected on the basis of spin-echo and time-of-flight MR findings, but multisection CT angiograms showed a patent sigmoid sinus and predominant drainage via the emissary veins toward the vertebral plexus, with only a minor contribution of the jugular veins. This case illustrates the variability of the venous anatomy in the craniocervical region.
Assuntos
Veias Cerebrais/anormalidades , Cavidades Cranianas/anormalidades , Adulto , Veias Cerebrais/diagnóstico por imagem , Cavidades Cranianas/diagnóstico por imagem , Diagnóstico Diferencial , Feminino , Humanos , Angiografia por Ressonância Magnética , Imageamento por Ressonância Magnética , Trombose dos Seios Intracranianos/diagnóstico , Tomografia Computadorizada por Raios X , Ultrassonografia , Vertigem/diagnósticoRESUMO
Triggers of brain inflammation in pneumococcal meningitis are unknown. TNF-α and IL-1ß were upregulated (real time PCR and in situ hybridization) in neurons and astrocytes time-dependently and maximally in the hippocampus during murine pneumococcal meningitis. Upregulation of TNF-α and IL-1ß mRNA in the brain parenchyma was independent of cerebrospinal fluid leukocytosis, pneumococcal pneumolysin and H2O2, but it was potently induced by pneumococcal cell wall (PCW) fragments. Brain TNF-α mRNA was downregulated by a matrix metalloproteinases inhibitor. PCW fragments were located in the brain parenchyma. In conclusion, PCW fragments and matrix metalloproteinases trigger cytokine induction in the brain parenchyma during pneumococcal meningitis.
Assuntos
Encéfalo/metabolismo , Encéfalo/patologia , Regulação da Expressão Gênica/fisiologia , Interleucina-1beta/metabolismo , Meningite Pneumocócica/patologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Anticorpos/farmacologia , Anticorpos/uso terapêutico , Encéfalo/microbiologia , Antígenos CD18/imunologia , Contagem de Colônia Microbiana , Dipeptídeos/farmacologia , Dipeptídeos/uso terapêutico , Modelos Animais de Doenças , Dura-Máter/metabolismo , Dura-Máter/patologia , Proteína Glial Fibrilar Ácida/metabolismo , Interleucina-1beta/genética , Masculino , Inibidores de Metaloproteinases de Matriz/uso terapêutico , Metaloproteinases da Matriz/metabolismo , Meningite Pneumocócica/tratamento farmacológico , Meningite Pneumocócica/etiologia , Camundongos , Camundongos Endogâmicos C57BL , Fosforilcolina/metabolismo , Streptococcus pneumoniae/patogenicidade , Fator de Necrose Tumoral alfa/genéticaAssuntos
Carcinoma/complicações , Hemorragia Cerebral/etiologia , Fibrinólise , Neoplasias da Próstata/complicações , Idoso , Idoso de 80 Anos ou mais , Carcinoma/diagnóstico , Hemorragia Cerebral/diagnóstico , Diagnóstico Diferencial , Fibrinólise/fisiologia , Humanos , Masculino , Neoplasias da Próstata/diagnósticoAssuntos
Circulação Cerebrovascular/fisiologia , Eclampsia/diagnóstico , Eclampsia/fisiopatologia , Adulto , Eclampsia/terapia , Feminino , Humanos , Recém-Nascido , Angiografia por Ressonância Magnética/métodos , Imageamento por Ressonância Magnética/métodos , Masculino , Gravidez , Ultrassonografia Doppler Dupla/métodos , Adulto JovemRESUMO
Caspases are important executioners of the endogenous cell death program. However, their function is not restricted to the induction of cell death. Caspases may process cytokines and contribute to cell differentiation or lymphocyte proliferation. In addition to their pleiotropic functions we show evidence that, under certain conditions, caspases are activated during apoptosis without executing the cell death program. Following whole body irradiation, p53 and caspases were activated in both the cerebellum and eye of postnatal day 5 mice. Although p53 activation and cell death kinetics were similar in both the cerebellum and eye, the processing of caspases was protracted and reduced in the eye. In particular, retinal caspase activation appeared not to be the executioner of cell death; incubation of retinal and cerebellar explants in the presence of the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone prevented DNA fragmentation, a hallmark of apoptosis, only in cerebellar granule cells. In contrast, in retinal cells no impairment of DNA fragmentation was observed in the presence of N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone, indicating p53-dependent but caspase-independent cell death pathways despite caspase activation.
Assuntos
Apoptose/efeitos da radiação , Fragmentação do DNA/efeitos da radiação , Radiação , Retina/citologia , Células-Tronco/efeitos da radiação , Proteína Supressora de Tumor p53/fisiologia , Animais , Animais Recém-Nascidos , Fator de Indução de Apoptose/metabolismo , Caspases/metabolismo , Cerebelo/efeitos da radiação , Relação Dose-Resposta à Radiação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Técnicas de Cultura de Órgãos , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/metabolismo , Proteína Supressora de Tumor p53/deficiênciaRESUMO
Bacterial toxins such as pneumolysin are key mediators of cytotoxicity in infections. Pneumolysin is a pore-forming toxin released by Streptococcus pneumoniae, the major cause of bacterial meningitis. We found that pneumolysin is the pneumococcal factor that accounts for the cell death pathways induced by live bacteria in primary neurons. The pore-forming activity of pneumolysin is essential for the induction of mitochondrial damage and apoptosis. Pneumolysin colocalized with mitochondrial membranes, altered the mitochondrial membrane potential, and caused the release of apoptosis-inducing factor and cell death. Pneumolysin induced neuronal apoptosis without activating caspase-1, -3, or -8. Wild-type pneumococci also induced apoptosis without activation of caspase-3, whereas pneumolysin-negative pneumococci activated caspase-3 through the release of bacterial hydrogen peroxide. Pneumolysin caused upregulation of X-chromosome-linked inhibitor of apoptosis protein and inhibited staurosporine-induced caspase activation, suggesting the presence of actively suppressive mechanisms on caspases. In conclusion, our results indicate additional functions of pneumolysin as a mitochondrial toxin and as a determinant of caspase-independent apoptosis. Considering this, blocking of pneumolysin may be a promising cytoprotective strategy in pneumococcal meningitis and other infections.
Assuntos
Mitocôndrias/microbiologia , Neurônios/microbiologia , Streptococcus pneumoniae/patogenicidade , Estreptolisinas/fisiologia , Animais , Apoptose/fisiologia , Proteínas de Bactérias/fisiologia , Cálcio/metabolismo , Morte Celular/fisiologia , Células Cultivadas , Potencial da Membrana Mitocondrial/fisiologia , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Membranas Mitocondriais/microbiologia , Membranas Mitocondriais/patologia , Neurônios/metabolismo , Neurônios/patologia , Ratos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
Innate immunity relies on pattern recognition receptors to detect the presence of infectious pathogens. In the case of Gram-positive bacteria, binding of bacterial lipopeptides to TLR2 is currently regarded as an important mechanism. In the present study, we used the synthetic bacterial lipopeptide Pam3CysSK4, a selective TLR2 agonist, to induce meningeal inflammation in rodents. In a 6-h rat model, intrathecal application of Pam3CysSK4 caused influx of leukocytes into the cerebrospinal fluid (CSF) and induced a marked increase of regional cerebral blood flow and intracranial pressure. In wild-type mice, we observed CSF pleocytosis and an increased number of apoptotic neurons in the dentate gyrus 24 h after intrathecal challenge. Inflammation and associated neuronal loss were absent in TLR2 knockout mice. In purified neurons, cytotoxicity of Pam3CysSK4 itself was not observed. Exposure of microglia to Pam3CysSK4 induced neurotoxic properties in the supernatant of wild-type, but not TLR2-deficient microglia. We conclude that TLR2-mediated signaling is sufficient to induce the host-dependent key features of acute bacterial meningitis. Therefore, synthetic lipopeptides are a highly specific tool to study mechanisms of TLR2-driven neurodegeneration in vivo.
Assuntos
Meningites Bacterianas/imunologia , Meningites Bacterianas/patologia , Receptor 2 Toll-Like/fisiologia , Animais , Células Cultivadas , Modelos Animais de Doenças , Injeções Espinhais , Lipopeptídeos , Masculino , Meningites Bacterianas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neuroglia/imunologia , Neuroglia/patologia , Neurônios/imunologia , Neurônios/patologia , Peptídeos/administração & dosagem , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/metabolismo , Infecções Pneumocócicas/patologia , Ratos , Ratos Wistar , Streptococcus pneumoniae/imunologia , Receptor 2 Toll-Like/agonistas , Receptor 2 Toll-Like/antagonistas & inibidores , Receptor 2 Toll-Like/deficiênciaAssuntos
Ambulâncias , Serviços Médicos de Emergência/métodos , Doenças do Sistema Nervoso/etiologia , Doenças do Sistema Nervoso/prevenção & controle , Fármacos Neuroprotetores/uso terapêutico , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/terapia , Isquemia Encefálica/complicações , Isquemia Encefálica/terapia , Fibrinolíticos/uso terapêutico , Humanos , Ativador de Plasminogênio Tecidual/uso terapêutico , Tomografia Computadorizada por Raios XRESUMO
New Zealand White rabbits were challenged with the wild-type (wt) group B streptococci (GBS) serotype III strain (COH1) and its isogenic nonhemolytic (NH) and hyperhemolytic (HH) mutants. Mortality differed significantly between rabbits infected with the HH mutant IN40 (67%), compared with rabbits infected with the wt COH1 strain (27%) and the NH strains COH1-20 and COH1:cylEDeltacat (13% and 0%, respectively; P<.05). Histopathologically, disseminated septic microabscesses surrounded by necrotic foci were found exclusively in the livers of HH mutant IN40-infected animals. Serum transaminase levels were 20-fold higher in the HH-infected group, compared with rabbits infected with the other strains. Positive TUNEL (in situ terminal deoxynucleotide transferase-mediated dUTP nick end labeling) staining and activation of caspase-3 in hepatocytes were more frequent in HH-infected than in wt-infected animals and absent in the NH mutant COH1-20-infected group, indicating that GBS beta-hemolysin triggers apoptotic pathways in hepatocytes. This work provides the first evidence that GBS beta-hemolysin plays a crucial role in the pathophysiology of GBS sepsis by inducing liver failure and high mortality.
Assuntos
Proteínas Hemolisinas/toxicidade , Fígado/efeitos dos fármacos , Choque Séptico/mortalidade , Animais , Proteínas de Bactérias , Pressão Sanguínea/efeitos dos fármacos , Caspase 3 , Caspases/metabolismo , Ativação Enzimática , Marcação In Situ das Extremidades Cortadas , Coelhos , Streptococcus agalactiae , Transaminases/sangueRESUMO
Radiation-induced death in the developing brain is p53-dependent. However, genetic studies indicate that the signalling pathways that couple irradiation to p53 expression can vary between different developing neural populations [Herzog et al. (1998) Science, 280, 1089-1091]. Here we establish that signalling downstream of p53 also exhibits brain region-specific differences that are associated with the relative vulnerability of some cell populations to radiation-induced killing in the mouse. Following gamma-irradiation, p53 and p21WAF1/cip1, but not Bax, protein levels increased in the developing cerebellum. In contrast, neither p21WAF1/cip1 nor Bax protein levels were elevated in the retina following irradiation, despite increased p53 expression. In the retina, p53 expression was associated with cells destined to die, whereas in the cerebellum, p53 was expressed in both radiation-sensitive and radiation-resistant neuroblasts of the external granule cell layer. Although p21WAF1/cip1 mRNA was expressed in all p53-positive neuroblasts after irradiation, p21WAF1/cip1 protein was only detected in radiation-resistant neuroblasts of the cerebellum. Thus, p21WAF1/cip1 was subject to post-transcriptional regulation with p21WAF1/cip1 protein only accumulating in cells destined to survive irradiation. Nevertheless, p21WAF1/cip1 function was not essential for radiation resistance, as postmitotic neuroblasts in the external granule cell layer were spared in p21WAF1/cip1 knockout mice.
Assuntos
Morte Celular/efeitos da radiação , Sistema Nervoso Central/efeitos da radiação , Ciclinas/efeitos da radiação , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Neurônios/efeitos da radiação , Proteínas Proto-Oncogênicas c-bcl-2 , Transcrição Gênica/efeitos da radiação , Animais , Animais Recém-Nascidos , Morte Celular/genética , Divisão Celular/genética , Divisão Celular/efeitos da radiação , Sobrevivência Celular/genética , Sobrevivência Celular/efeitos da radiação , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/metabolismo , Cerebelo/crescimento & desenvolvimento , Cerebelo/metabolismo , Cerebelo/efeitos da radiação , Inibidor de Quinase Dependente de Ciclina p21 , Ciclinas/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Masculino , Camundongos , Camundongos Knockout , Neurônios/metabolismo , Neurônios/patologia , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas/efeitos da radiação , Retina/crescimento & desenvolvimento , Retina/metabolismo , Retina/efeitos da radiação , Transcrição Gênica/fisiologia , Proteína Supressora de Tumor p53/deficiência , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/efeitos da radiação , Proteína X Associada a bcl-2RESUMO
Significant injury during bacterial meningitis arises from mechanisms of neuronal apoptosis, particularly in the hippocampus. Apoptosis can involve both the caspase-dependent and the caspase-independent pathway, and, although both pathways have been implicated in pneumococcus-induced neuronal cell death, their relative contributions in vivo are unclear. We used mice deficient in the activation of caspase-3, ATM, and p53 to examine the role that caspase-dependent apoptosis plays in neuronal death in the context of pneumococcal meningitis. The overall symptomatology of acute infection was similar in all mice tested, indicating that late sequelae are the clinical manifestations of neuronal death. Two phases of apoptosis were discernible: neuronal injury at 18 h after infection was independent of the caspase-3 pathway, and neuronal cell death at 24 h after infection was attenuated in the absence of the caspase-3 pathway. We conclude that treatments to increase the survival rate of neurons in patients with meningitis will need to take into account at least these 2 mechanisms of damage.
Assuntos
Apoptose , Caspases/fisiologia , Hipocampo/fisiopatologia , Meningite Pneumocócica/fisiopatologia , Animais , Proteínas Mutadas de Ataxia Telangiectasia , Caspase 3 , Caspases/deficiência , Caspases/genética , Proteínas de Ciclo Celular , Proteínas de Ligação a DNA , Modelos Animais de Doenças , Camundongos , Camundongos Knockout , Neurônios/patologia , Neurônios/fisiologia , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Fatores de Tempo , Proteína Supressora de Tumor p53/deficiência , Proteína Supressora de Tumor p53/genética , Proteínas Supressoras de TumorRESUMO
Gelsolin (gsn), an actin-severing protein, protects neurons from excitotoxic cell death via inactivation of membranous Ca(2+) channels. Its role during apoptotic cell death, however, has remained unclear. Using several models of neuronal cell death, we demonstrate that endogenous gelsolin has anti-apoptotic properties that correlate to its dynamic actions on the cytoskeleton. We show that neurons lacking gelsolin (gsn(-/-)) have enhanced apoptosis following exposure to staurosporine, thapsigargin, or the cholinergic toxin ethylcholine aziridinium (AF64A). AF64A-induced loss of mitochondrial membrane potential and activation of caspase-3 was specifically enhanced in gsn(-/-) neurons and could be reversed by pharmacological inhibition of mitochondrial permeability transition. Moreover, increased caspase-3 activation and cell death in AF64A-treated gsn(-/-) neurons were completely reversed by pharmacological depolymerization of actin filaments and further enhanced by their stabilization. In conclusion, actin remodeling by endogenous gelsolin or analogues protects neurons from apoptosis mediated by mitochondria and caspase-3.