RESUMO
BACKGROUND: Acute intoxication with organophosphorus (OP) cholinesterase inhibitors can trigger convulsions that progress to life-threatening status epilepticus. Survivors face long-term morbidity including mild-to-severe decline in memory. It is posited that neuroinflammation plays a key role in the pathogenesis of OP-induced neuropsychiatric deficits. Rigorous testing of this hypothesis requires preclinical models that recapitulate relevant phenotypic outcomes. Here, we describe a rat model of acute intoxication with the OP diisopropylfluorophosphate (DFP) that exhibits persistent neuroinflammation and cognitive impairment. METHODS: Neuroinflammation, neurodegeneration, and cognitive function were compared in adult male Sprague Dawley rats injected with an acutely toxic dose of DFP vs. vehicle controls at multiple time points up to 36 days post-exposure. Neuroinflammation was quantified using immunohistochemical biomarkers of microglia (ionized calcium-binding adapter molecule 1, IBA1) and activated astrocytes (glial fibrillary acidic protein, GFAP) and positron emission tomography (PET) imaging of [11C]-(R)-PK11195, a ligand for the 18-kDa mitochondrial membrane translocator protein (TSPO). FluoroJade-B staining was used to assess neurodegeneration; Pavlovian conditioning, to assess cognitive function. RESULTS: Animals exhibited moderate-to-severe seizures within minutes of DFP injection that continued for up to 6 h post-injection. As indicated by IBA1 and GFAP immunoreactivity and by PET imaging of TSPO, acute DFP intoxication triggered neuroinflammation in the hippocampus and cortex during the first 3 days that peaked at 7 days and persisted to 21 days post-exposure in most animals. Neurodegeneration was detected in multiple brain regions from 1 to 14 days post-exposure. All DFP-intoxicated animals exhibited significant deficits in contextual fear conditioning at 9 and 20 days post-exposure compared to vehicle controls. Whole-brain TSPO labeling positively correlated with seizure severity score, but did not correlate with performance in the contextual fear-conditioning task. CONCLUSIONS: We describe a preclinical model in which acute DFP intoxication causes seizures, persistent neuroinflammation, neurodegeneration, and memory impairment. The extent of the neuroinflammatory response is influenced by seizure severity. However, the observation that a subset of animals with moderate seizures and minimal TSPO labeling exhibited cognitive deficits comparable to those of animals with severe seizures and significant TSPO labeling suggests that DFP may impair learning and memory circuitry via mechanisms independent of seizures or neuroinflammation.
Assuntos
Inibidores da Colinesterase/toxicidade , Disfunção Cognitiva/induzido quimicamente , Encefalite/induzido quimicamente , Isoflurofato/toxicidade , Animais , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Transporte/metabolismo , Disfunção Cognitiva/diagnóstico por imagem , Condicionamento Clássico/efeitos dos fármacos , Encefalite/diagnóstico por imagem , Comportamento Exploratório/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Imageamento por Ressonância Magnética , Masculino , Proteínas dos Microfilamentos/metabolismo , Tomografia por Emissão de Pósitrons , Ratos , Ratos Sprague-Dawley , Receptores de GABA-A/metabolismo , Análise de Regressão , Fatores de TempoRESUMO
Tetramethylenedisulfotetramine (TETS) is a potent convulsant poison for which there is currently no approved antidote. The convulsant action of TETS is thought to be mediated by inhibition of type A gamma-aminobutyric acid receptor (GABAAR) function. We, therefore, investigated the effects of post-exposure administration of diazepam, a GABAAR positive allosteric modulator, on seizure activity, death and neuroinflammation in adult male Swiss mice injected with a lethal dose of TETS (0.15mg/kg, ip). Administration of a high dose of diazepam (5mg/kg, ip) immediately following the second clonic seizure (approximately 20min post-TETS injection) effectively prevented progression to tonic seizures and death. However, this treatment did not prevent persistent reactive astrogliosis and microglial activation, as determined by GFAP and Iba-1 immunoreactivity and microglial cell morphology. Inhibition of soluble epoxide hydrolase (sEH) has been shown to exert potent anti-inflammatory effects and to increase survival in mice intoxicated with other GABAAR antagonists. The sEH inhibitor TUPS (1mg/kg, ip) administered immediately after the second clonic seizure did not protect TETS-intoxicated animals from tonic seizures or death. Combined administration of diazepam (5mg/kg, ip) and TUPS (1mg/kg, ip, starting 1h after diazepam and repeated every 24h) prevented TETS-induced lethality and influenced signs of neuroinflammation in some brain regions. Significantly decreased microglial activation and enhanced reactive astrogliosis were observed in the hippocampus, with no changes in the cortex. Combining an agent that targets specific anti-inflammatory mechanisms with a traditional antiseizure drug may enhance treatment outcome in TETS intoxication.
Assuntos
Anti-Inflamatórios/administração & dosagem , Anticonvulsivantes/administração & dosagem , Encéfalo/efeitos dos fármacos , Hidrocarbonetos Aromáticos com Pontes , Diazepam/administração & dosagem , Encefalite/prevenção & controle , Inibidores Enzimáticos/administração & dosagem , Epóxido Hidrolases/antagonistas & inibidores , Moduladores GABAérgicos/administração & dosagem , Compostos de Fenilureia/administração & dosagem , Piperidinas/administração & dosagem , Convulsões/prevenção & controle , Animais , Encéfalo/enzimologia , Encéfalo/fisiopatologia , Ondas Encefálicas/efeitos dos fármacos , Modelos Animais de Doenças , Esquema de Medicação , Quimioterapia Combinada , Eletroencefalografia , Encefalite/induzido quimicamente , Encefalite/enzimologia , Encefalite/fisiopatologia , Epóxido Hidrolases/metabolismo , Masculino , Camundongos , Convulsões/induzido quimicamente , Convulsões/enzimologia , Convulsões/fisiopatologia , Fatores de TempoRESUMO
Tetramethylenedisulfotetramine (tetramine; TETS) is a potent convulsant poison that is considered to be a chemical threat agent. To provide a basis for the investigation of antidotes for TETS-induced seizures, we characterized the convulsant activity of TETS in mice and rats when administered by the intraperitoneal, intravenous, oral, and intraventricular routes as a single acute dose and with repeated sublethal doses. In mice, parenteral and oral TETS caused immobility, myoclonic body jerks, clonic seizures of the forelimbs and/or hindlimbs, tonic seizures, and death. The CD50 values for clonic and tonic seizures after oral administration were 0.11 and 0.22 mg/kg, respectively. Intraventricular administration of TETS (5-100 µg) in rats also caused clonic-tonic seizures and death. In mice, repeated sublethal doses of TETS at intervals of 2, 24, and 48 h failed to result in the development of persistent enhanced seizure responsivity ("kindling") as was observed with repeated pentylenetetrazol treatment. In mice, sublethal doses of TETS that produced clonic seizures did not cause observable structural brain damage as assessed with routine histology and Fluoro-Jade B staining 7 days after treatment. However, 1 to 3 days after a single convulsant dose of TETS the expression of glial fibrillary acidic protein, an astrocyte marker, and ionized calcium binding adaptor molecule 1, a microglia marker, were markedly increased in cortex and hippocampus. Although TETS doses that are compatible with survival are not associated with overt evidence of cellular injury or neurodegeneration, there is transient reactive astrocytosis and microglial activation, indicating that brain inflammatory responses are provoked.
Assuntos
Hidrocarbonetos Aromáticos com Pontes/toxicidade , Convulsivantes/toxicidade , Convulsões/induzido quimicamente , Convulsões/metabolismo , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Extremidades , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/induzido quimicamente , Gliose/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Camundongos , Proteínas dos Microfilamentos/metabolismo , Microglia/efeitos dos fármacos , Microglia/metabolismo , Pentilenotetrazol/farmacologia , Picrotoxina/efeitos adversos , Ratos , Ratos Sprague-DawleyRESUMO
The emerald jewel wasp, Ampulex compressa, exploits the American cockroach, Periplaneta americana, as a host for its progeny. The wasp subdues the host by stinging directly into the brain and subesophageal ganglion, inducing long-term hypokinesia. The hypokinesic host lacks normal escape behavior and motivation to walk, making it easy for subjugation by the wasp. The mechanism underlying hypokinesia induction is not known, but depletion of monoamines induces behavior resembling venom-induced hypokinesia. To test whether amine depletion occurs in stung animals, we used high-performance liquid chromatography with electrochemical detection (HPLC-ED) to measure quantitatively amine levels in the central nervous system. Our data show clearly that levels of dopamine, serotonin, octopamine and tyramine remain unchanged in stung animals, whereas animals treated with reserpine exhibited marked depletion of all amines sampled. Furthermore, stung animals treated with reserpine show depletion of amines, demonstrating that envenomation also does not interfere with amine release. These results show that hypokinesia induced by Ampulex venom does not result from amine depletion or inability to release monoamines in the central nervous system.
Assuntos
Aminas Biogênicas/análise , Sistema Nervoso Central/efeitos dos fármacos , Periplaneta/química , Venenos de Vespas/metabolismo , Animais , Cromatografia Líquida de Alta Pressão/métodos , Dopamina/sangue , Interações Hospedeiro-Parasita , Himenópteros/química , Hipocinesia/induzido quimicamente , Hipocinesia/patologia , Masculino , Octopamina/sangue , Periplaneta/parasitologia , Reserpina/uso terapêutico , Serotonina/sangue , Tiramina/sangueRESUMO
Organophosphorus (OP) nerve agents and pesticides inhibit acetylcholinesterase (AChE), and this is thought to be a primary mechanism mediating the neurotoxicity of these compounds. However, a number of observations suggest that mechanisms other than or in addition to AChE inhibition contribute to OP neurotoxicity. There is significant experimental evidence that acute OP intoxication elicits a robust inflammatory response, and emerging evidence suggests that chronic repeated low-level OP exposure also upregulates inflammatory mediators. A critical question that is just beginning to be addressed experimentally is the pathophysiologic relevance of inflammation in either acute or chronic OP intoxication. The goal of this article is to provide a brief review of the current status of our knowledge linking inflammation to OP intoxication, and to discuss the implications of these findings in the context of therapeutic and diagnostic approaches to OP neurotoxicity.