RESUMEN
Pesticides are used in large amounts in agriculture and the evaluation of their toxic effects is of major concern to public and environmental health. The aim of the present study was to investigate the genotoxic potential of a commercial formulation of the fungicide mancozeb by the micronucleus test in bone marrow and the comet assay in total blood of Wistar rats. Adult male Wistar rats were treated with a solution of mancozeb at a concentration of 40 mg/kg/day, administered intraperitoneally for 18 consecutive days, and compared to a control group. The results indicate that mancozeb induced significantly higher DNA damage as detected by the comet assay and increased the frequency of micronuclei. The results show that mancozeb is genotoxic and may adversely affect the DNA integrity of exposed organisms.
Asunto(s)
Daño del ADN , Ditiocarba/toxicidad , Maneb/toxicidad , Zineb/toxicidad , Animales , Recuento de Eritrocitos , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Masculino , Micronúcleos con Defecto Cromosómico/efectos de los fármacos , Pruebas de Micronúcleos , Ratas WistarRESUMEN
A planta Salvia divinorum Epling & Játiva (SDI), da família Lamiaceae, tem sido usada por séculos pela cultura mazateca e vem ganhando popularidade como droga recreacional nos últimos anos. Seu princípio ativo - Salvinorina A (SA) - é agonista dos receptores opióides kappa, com potencial psicotrópico. A utilização da planta vem crescendo na Europa e na América do Norte, apesar de ainda não existirem provas concretas sobre abuso. A presente revisão da literatura contemporânea aborda as evidências sobre o potencial de abuso de SDI, bem como o crescente uso recreacional, ainda que seja alucinógeno permitido legalmente e de fácil compra em muitos países.
The plant Salvia divinorum Epling & Játiva (SDI), of the Lamiaceae family, has been used for centuries by the Mazateca culture and has gained popularity as a recreational drug in the last years. Its active principle, Salvinorin A (SA), is a potentially psychotropic agonist of the kappa opioid receptors. The use of SDI has increased in Europe and North America, although there are no concrete proofs about abuse. The present review discusses current evidence on potential SDI abuse, as well as its increasing recreational use, although it is considered a legalized hallucinogen easily acquired in many countries.
Asunto(s)
Alucinógenos , Trastornos Relacionados con Opioides , Recreación , Salvia , EpidemiologíaRESUMEN
The aim of this study was to determine the feasibility of combining water quality analysis with different biomarkers to characterise the relationship between anthropogenic contamination and biotic response in the Sinos River, southern Brazil. Wistar rats were studied using three biomarkers combined with physical, chemical and microbiological analysis to assess the effects of pollution at four sampling sites. The induction of oxidative stress was quantified by MDA levels in peripheral blood, lymphocyte DNA damage was determined using the comet assay, and histopathological changes were analysed in the liver. After sampling, animals were allowed to drink the river water during a 48 hours period. No increase in oxidative stress and DNA damage was observed. However, liver damage was observed in the animals exposed to water samples, indicating that the Sinos River is contaminated with hepatotoxic substances. Water analyses confirmed that water quality decreased downriver.
Asunto(s)
Daño del ADN/efectos de los fármacos , Monitoreo del Ambiente/métodos , Hígado/efectos de los fármacos , Malondialdehído/sangre , Ríos/química , Contaminantes Químicos del Agua/toxicidad , Animales , Biomarcadores/sangre , Brasil , Ensayo Cometa , Hígado/patología , Masculino , Estrés Oxidativo , Ratas , Ratas WistarAsunto(s)
Anticuerpos Antibacterianos/sangre , Infecciones por Chlamydophila/veterinaria , Chlamydophila/inmunología , Pruebas de Fijación del Complemento/veterinaria , Enfermedades de las Cabras/diagnóstico , Enfermedades de las Ovejas/diagnóstico , Animales , Infecciones por Chlamydophila/diagnóstico , Pruebas de Fijación del Complemento/métodos , Enfermedades de las Cabras/microbiología , Cabras , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Ovinos , Enfermedades de las Ovejas/microbiologíaRESUMEN
The aim of the present work was to test the role of protein tyrosine kinases (PTKs) on both the short-term memory (STM) and long-term memory (LTM) of the inhibitory avoidance task in rats using the inhibitor of tyrosine kinase, radicicol. Rats implanted with cannulae in the CA1 area of the dorsal hippocampus received a 0.5 microl infusion of radicicol (0.5, 1, 5, 10, 20 microg/ml) or vehicle (water) at different times after training and were tested for STM (1.5 or 3 h) and LTM (24 h). Additionally, one group received radicicol 10 min prior to the test for LTM. Radicicol depressed both STM and LTM when infused before and immediately after training and had no effect on either form of memory when infused 30 or 90 min after training. Radicicol also depressed the retrieval of LTM. Our results indicate that memory formation and retrieval in the hippocampus can involve PTK activity, but the present findings should be taken merely as a possible starting point for future investigations.
Asunto(s)
Reacción de Prevención/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Hipocampo/efectos de los fármacos , Lactonas/farmacología , Memoria a Corto Plazo/efectos de los fármacos , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Retención en Psicología/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Electrochoque , Macrólidos , Masculino , Ratas , Ratas WistarRESUMEN
The cAMP/cAMP-dependent protein kinase (PKA) signaling pathway has been implicated in synaptic plasticity changes and memory consolidation. Several cortical structures are involved in the consolidation of memory for inhibitory avoidance. The aim of the present work was to observe the effects of training in the inhibitory avoidance task on the levels of PKA activity in the entorhinal, parietal and posterior cingulate cortex (EC, PARIET and PC), and the medial precentral area (Fr2) of the rat, at different post-training times (0, 1.5, 3 and 6h). PKA activity, assayed using [gamma-32P]ATP and kemptide, a selective substrate, increased in the EC 3 h after training, but no changes were observed in PARIET, PC and Fr2. These results suggest that the late phase of memory consolidation of inhibitory avoidance requires a functional PKA signaling pathway in the EC in a way that a 'peak' of PKA activity is observed.
Asunto(s)
Reacción de Prevención/fisiología , Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Corteza Entorrinal/fisiología , Inhibición Psicológica , Transducción de Señal/fisiología , Animales , Mapeo Encefálico , Corteza Cerebral/fisiología , Giro del Cíngulo/fisiología , Masculino , Lóbulo Parietal/fisiología , Ratas , Ratas WistarRESUMEN
RATIONALE: The generic antagonist of glutamate metabotropic receptors (mGlus), MCPG, blocks retrieval of inhibitory avoidance when infused into the CA1 area of rat hippocampus. It was considered important to study the effect of agonists of different types of mGlus on retrieval both of this task and of a related one, contextual fear. OBJECTIVES: To measure the effect of three mGlu agonists (3HPG, which is selective to mGlu1; LCCG, which binds to mGlu2 and mGlu3; and LAP-4, which binds to mGlu4 and mGlu6), infused bilaterally into CA1, on the retrieval of one-trial inhibitory avoidance and contextual fear in rats. METHODS: Rats bilaterally implanted with cannulae in the CA1 region of the dorsal hippocampus were trained in one-trial step-down inhibitory avoidance or in a contextual fear task and tested for retention 24 h later. The drugs 3HPG, LCCG and LAP-4 were infused into CA1 at different concentration levels 10 min before retention testing. In addition, we studied the effect of these drugs on locomotor and exploratory activity measured in an open field, and on pro- and anti-conflict behaviour in an elevated plus-maze. RESULTS: 3HPG hindered, and LCCG and LAP-4 enhanced, retrieval of the two tasks. In all cases the effects were dose-dependent. The drugs had no effects on open field or plus maze behaviour. CONCLUSIONS: Retrieval of one-trial inhibitory avoidance and of contextual fear is regulated by mGlus in the CA1area of the rat hippocampus. The results suggest that mGlu2s, mGlu3s, mGlu4s and mGlu6s are necessary for retrieval and that mGlu1s play an inhibitory role. The effects are not explainable by nonspecific influences on locomotor or exploratory activity or anxiety levels.
Asunto(s)
Reacción de Prevención/efectos de los fármacos , Agonistas de Aminoácidos Excitadores/administración & dosificación , Glicina/análogos & derivados , Hipocampo/efectos de los fármacos , Inhibición Psicológica , Receptores de Glutamato Metabotrópico/agonistas , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Facilitación Social , Aminoácidos Dicarboxílicos/administración & dosificación , Animales , Reacción de Prevención/fisiología , Inhibidores Enzimáticos/administración & dosificación , Miedo/efectos de los fármacos , Miedo/fisiología , Glicina/administración & dosificación , Hipocampo/fisiología , Inyecciones Intraventriculares , Masculino , Ratas , Ratas Wistar , Receptores de Glutamato Metabotrópico/fisiologíaRESUMEN
Retrieval of inhibitory avoidance has been recently shown to require intact glutamate receptors, protein kinases A and C and mitogen-activated protein kinase in the CA1 region of the rat hippocampus and in the entorhinal, posterior parietal and anterior cingulate cortex. These enzymatic activities are known to be modulated by dopamine D(1), beta-noradrenergic, 5HT1A and cholinergic muscarinic receptors. Here we study the effect on retrieval of this task of well-known agonists and antagonists of these receptors infused in the same brain cortical regions and into the basolateral amygdala, in rats. The drugs used were SKF38393 (D(1) agonist), noradrenaline, 8-HO-DPAT (5HT1A agonist), oxotremorine (muscarinic agonist), SCH23390 (D(1) antagonist), timolol (beta antagonist), NAN-190 (5HT1A antagonist) and scopolamine (muscarinic antagonist). All were studied at two different dose levels. The localised infusion of SKF38393, noradrenaline, NAN-190 and oxotremorine into any of the cortical structures mentioned 10 min prior to a 24-h retention test session of one-trial step-down inhibitory avoidance enhanced retention test performance. SCH2330, timolol, 8-HO-DPAT and scopolamine hindered retention test performance. In the basolateral amygdala only an enhancing effect of noradrenaline and an inhibitory effect of timolol were seen. Three hours after the infusions, retention test performance returned to normal in all cases. None of the treatments affected locomotion or rearing in an open field or behaviour in the elevated plus maze. Therefore, their effects on retention testing can be attributed to an influence on retrieval. In conclusion, memory retrieval of this apparently simple task requires the participation of CA1, entorhinal, posterior parietal and anterior cingulate cortex, and is strongly modulated by, dopaminergic D(1), beta-noradrenergic, muscarinic cholinergic and 5HT1A receptors in the four areas. The first three types of receptor enhance, and the latter inhibits, retrieval. Only beta-adrenoceptors appears to be involved in the modulation of retrieval of this task by the amygdala. The results bear on the well-known influence of emotion and mood on retrieval, and indicate that this involves many areas of the brain simultaneously. In addition, the results point to similarities and differences between the modulatory mechanisms that affect retrieval and those involved in the consolidation of the same task.
Asunto(s)
Reacción de Prevención/fisiología , Corteza Cerebral/fisiología , Receptores Adrenérgicos/fisiología , Receptores de Dopamina D1/fisiología , Receptores Muscarínicos/fisiología , Receptores de Serotonina/fisiología , Retención en Psicología/fisiología , Amígdala del Cerebelo/fisiología , Animales , Nivel de Alerta/fisiología , Mapeo Encefálico , Corteza Entorrinal/fisiología , Giro del Cíngulo/fisiología , Hipocampo/fisiología , Aprendizaje por Laberinto/fisiología , Recuerdo Mental/fisiología , Inhibición Neural/fisiología , Ratas , Receptores de Serotonina 5-HT1RESUMEN
Rats were implanted bilaterally with cannulae in the CA1 region of the dorsal hippocampus, the entorhinal cortex, anterior cingulate cortex, posterior parietal cortex, or the basolateral complex of the amygdala. The animals were trained in one-trial step-down inhibitory avoidance and tested 24 h later. Prior (10 min) to the retention test, through the cannulae, they received 0.5 microl infusions of a vehicle (2% dimethylsulfoxide in saline), or of the following drugs dissolved in the vehicle: the glutamate NMDA receptor blocker, aminophosphonopentanoic acid (AP5, 2.0 or 5.0 microg), the AMPA receptor blocker, 6,7-dinitroquinoxaline-2,3 (1H,4H)dione (DNQX, 0.4 or 1.0 microg), the metabotropic receptor antagonist, methylcarboxyphenylglycine (MCPG, 0.5 or 2.5 microg), the inhibitor of cAMP-dependent protein kinase (PKA), Rp-cAMPs (0.1 or 0.5 microg), the PKA stimulant, Sp-cAMPs (0.5 microg), or the inhibitor of the mitogen-activated protein kinase (MAPK), PD098059 (10 or 50 microM). All these drugs, at the same doses, had been previously found to alter long-term memory formation of this task. Here, retrieval test performance was blocked by DNQX, MCPG, Rp-cAMPs and PD098059 and enhanced by Sp-cAMPs infused into CA1 or the entorhinal cortex. The drugs had similar effects when infused into the parietal or anterior cingulate cortex, except that in these two areas AP5 also blocked retrieval, and in the cingulate cortex DNQX had no effect. Infusions into the basolateral amygdala were ineffective except for DNQX, which hindered retrieval. None of the treatments that affected retrieval had any influence on performance in an open field or in a plus maze; therefore, their effect on retention testing can not be attributed to an influence on locomotion, exploration or anxiety. The results indicate that the four cortical regions studied participate actively in, and are necessary for, retrieval of the one-trial avoidance task. They require metabotropic and/or NMDA glutamate receptors and PKA and MAPK activity. In contrast, the basolateral amygdala appears to participate only through a maintenance of its regular excitatory transmission mediated by glutamate AMPA receptors.
Asunto(s)
Reacción de Prevención/fisiología , Corteza Cerebral/fisiología , Antagonistas de Aminoácidos Excitadores/farmacología , Transducción de Señal/fisiología , 2-Amino-5-fosfonovalerato/administración & dosificación , 2-Amino-5-fosfonovalerato/farmacología , Adenilil Ciclasas/metabolismo , Animales , Reacción de Prevención/efectos de los fármacos , Benzoatos/administración & dosificación , Benzoatos/farmacología , Corteza Cerebral/efectos de los fármacos , Proteínas Quinasas Dependientes de AMP Cíclico/antagonistas & inhibidores , Corteza Entorrinal/fisiología , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/farmacología , Antagonistas de Aminoácidos Excitadores/administración & dosificación , Lateralidad Funcional/fisiología , Glicina/administración & dosificación , Glicina/análogos & derivados , Glicina/farmacología , Hipocampo/fisiología , Inyecciones , Quinoxalinas/administración & dosificación , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Receptores AMPA/antagonistas & inhibidores , Receptores de Glutamato Metabotrópico/antagonistas & inhibidores , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacosRESUMEN
The ability to recall past events is a major determinant of survival strategies in all species and is of paramount importance in determining our uniqueness as individuals. In contrast to memory formation, the information about the molecular mechanisms of memory retrieval is surprisingly scarce and fragmentary. Here we show that pretest inhibition of the specific upstream activator of mitogen-activated protein kinase kinase, or of protein kinase A in the hippocampus, blocked retrieval of long-term memory for an inhibitory avoidance task, a hippocampal-dependent learning task. An activator of protein kinase A enhanced retrieval. Mitogen-activated protein kinase activation increased in the hippocampus during retrieval, while protein kinase A activity remained unchanged. Pretest intrahippocampal blockade of metabotropic glutamate receptors or alpha-amino-3-hydroxy-5-methyl-4-isoxazolone propionic acid/kainate receptors, but not N-methyl-D-aspartate receptors or calcium/calmodulin dependent-protein kinase II, impaired retrieval. Thus, recall of inhibitory avoidance activates mitogen-activated protein kinase, which is necessary, along with metabotropic glutamate receptors, alpha-amino-3-hydroxy-5-methyl-4-isoxazolone propionic acid/kainate receptors, and protein kinase A, for long-term memory expression. Our results indicate that memory formation and retrieval may share some molecular mechanisms in the hippocampus.
Asunto(s)
Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Hipocampo/fisiología , Recuerdo Mental/fisiología , Proteínas Quinasas Activadas por Mitógenos/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Animales , Reacción de Prevención/fisiología , Activación Enzimática , Masculino , Ratas , Ratas WistarRESUMEN
One-trial step-down inhibitory (passive) avoidance training is followed by two peaks of cAMP-dependent protein kinase (PKA) activity in rat CA1: one immediately after training and the other 3 h later. The second peak relies on the first: Immediate posttraining infusion into CA1 of the inhibitor of the regulatory subunit of PKA, Rp-cAMPS, at a dose that reduces PKA activity during less than 90 min, cancelled both peaks. Long-term memory (LTM) of this task measured at 24 h depends on the two peaks: Rp-cAMPS given into CA1 0 or 175 min posttraining, but not between those times, blocked LTM. However, the effect of immediate posttraining Rp-cAMPS on LTM could not be reversed by the activator of the regulatory subunit of PKA, Sp-cAMPS, given at 180 min, which suggests that, for LTM, the first peak may be more important than the second. When given at 0, 22, 45, or 90, but not at 175 min from training, Rp-cAMPS blocked short-term memory (STM) measured at 90 or 180 min. This effect of immediate posttraining Rp-cAMPS infusion on STM but not that on LTM was readily reversed by Sp-cAMPS infused 22 min later. On its own, Sp-cAMPS had effects exactly opposite to those of the inhibitor. It enhanced LTM when given at 0 or 175 min from training, and it enhanced STM when given at 0, 22, 45, or 90 min from training. These findings show that STM and LTM formation require separate PKA-dependent processes in CA1. STM relies on the continued activity of the enzyme during the first 90 min. LTM relies on the two peaks of PKA activity that occur immediately and 180 min posttraining.
Asunto(s)
Reacción de Prevención/fisiología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , AMP Cíclico/análogos & derivados , Hipocampo/fisiología , Memoria a Corto Plazo/fisiología , Memoria/fisiología , Tionucleótidos/farmacología , Animales , Reacción de Prevención/efectos de los fármacos , AMP Cíclico/administración & dosificación , AMP Cíclico/farmacología , Electrochoque , Inhibidores Enzimáticos/administración & dosificación , Inhibidores Enzimáticos/farmacología , Hipocampo/efectos de los fármacos , Hipocampo/enzimología , Infusiones Parenterales , Memoria/efectos de los fármacos , Memoria a Corto Plazo/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/enzimología , Neuronas/fisiología , Ratas , Ratas Wistar , Tionucleótidos/administración & dosificación , Factores de TiempoRESUMEN
Rats were trained in one-trial step-down inhibitory avoidance and tested either 3 h or 31 days later. Ten minutes prior to the retention test, through indwelling cannulae placed in the CA1 region of the dorsal hippocampus, they received 0.5 microl infusions of: saline, a vehicle (2% dimethylsulfoxide in saline), the glutamate NMDA receptor blocker, aminophosphonopentanoic acid (AP5) (5.0 microg), the AMPA/kainate receptor blocker, cyanonitroquinoxaline dione (CNQX) (0.25 or 1.25 microg), the metabotropic receptor antagonist, methylcarboxyphenylglycine (MCPG) (0.5 or 2.5 microg), the inhibitor of calcium/calmodulin-dependent protein kinase II (KN62) (3.5 microg), the inhibitor of cAMP-dependent protein kinase (PKA), Rp-cAMPs (0.1 or 0.5 microg), the stimulant of the same enzyme, Sp-cAMPs (0.1 or 0.5 microg), or the inhibitor of the mitogen-activated protein kinase (MAPK) kinase, PD098059 (10 or 50 microM). CNQX, KN62 and PD098059 were dissolved in the vehicle; the other drugs were dissolved in saline. All these drugs, at the same doses, had been previously found to affect short- and long-term memory formation of this task. Retrieval measured 3 h after training (short-term memory) was blocked by CNQX and MCPG, and was unaffected by all the other drugs. In contrast, retrieval measured at 31 days was blocked by MCPG, Rp-cAMPs and PD098059, enhanced by Sp-cAMPs, and unaffected by CNQX, AP5 or KN62. The results indicate that, in CA1, glutamate metabotropic receptors are necessary for the retrieval of both short- and long-term memory; AMPA/kainate receptors are necessary for short-term but not long-term memory retrieval, and NMDA receptors are uninvolved in retrieval. Both the PKA and MAPK signalling pathways are required for the retrieval of long-term but not short-term memory.
Asunto(s)
Reacción de Prevención/fisiología , Miedo/fisiología , Hipocampo/fisiología , Memoria a Corto Plazo/fisiología , Receptores de Neurotransmisores/fisiología , Retención en Psicología/fisiología , Transmisión Sináptica/fisiología , Animales , Mapeo Encefálico , Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Proteínas Quinasas Activadas por Mitógenos/fisiología , Ratas , Ratas Wistar , Receptores AMPA/fisiología , Receptores de Ácido Kaínico/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Receptores de N-Metil-D-Aspartato/fisiologíaRESUMEN
Rats were implanted with cannulae in the CA1 area of the dorsal hippocampus or in the entorhinal cortex and trained in one-trial step-down inhibitory avoidance. Two retention tests were carried out in each animal, one at 1.5 h to measure short-term memory (STM) and another at 24 h to measure long-term memory (LTM). The purpose of the present study was to screen the effect on STM of various drugs previously shown to affect LTM of this task when given posttraining at the same doses that were used here. The drugs and doses were the guanylyl cyclase inhibitor LY83583 (LY, 2.5 microMg), the inhibitor of Tyr-protein kinase at low concentrations and of protein kinase G (PKG) at higher concentrations lavendustin A (LAV, 0.1 and 0.5 microMg), the PKG inhibitor KT5823 (2.0 microMg), the protein kinase C (PKC) inhibitor staurosporin (STAU, 2.5 microMg), the inhibitor of calcium/ calmodulin protein kinase II (CaMKII) KN62 (3.6 microMg), the protein kinase A (PKA) inhibitor KT5720 (0.5 microMg), and the mitogen-activated protein kinase kinase (MAPKK) inhibitor PD098059 (PD, 0.05 microMg). PD was dissolved in saline; all the other drugs were dissolved in 20% dimethyl sulfoxide. In all cases the drugs affected LTM as had been described in previous papers. The drugs affected STM and LTM differentially depending on the brain structure into which they were infused. STM was inhibited by KT5720, LY, and PD given into CA1 and by STAU and KT5720 given into the entorhinal cortex. PD given into the entorhinal cortex enhanced STM. LTM was inhibited by STAU, KN62, KT5720, KT5823, and LAV (0.5 microMg) given into CA1 and by STAU, KT5720, and PD given into the entorhinal cortex. The results suggest that STM and LTM involve different physiological mechanisms but are to an extent linked. STM appears to require PKA, guanylyl cyclase, and MAPKK activity in CA1 and PKA and PKC activity in the entorhinal cortex; MAPKK seems to play an inhibitory role in STM in the entorhinal cortex. In contrast, LTM appears to require PKA and PKC activity in both structures, guanylyl cyclase, PKG, and CaMKII activity in CA1, and MAPKK activity in the entorhinal cortex.
Asunto(s)
Carbazoles , Corteza Entorrinal/metabolismo , Inhibidores Enzimáticos/administración & dosificación , Hipocampo/metabolismo , Memoria/efectos de los fármacos , Memoria/fisiología , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/administración & dosificación , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados , Alcaloides/administración & dosificación , Aminoquinolinas/administración & dosificación , Animales , Reacción de Prevención/efectos de los fármacos , Corteza Entorrinal/efectos de los fármacos , Corteza Entorrinal/enzimología , Corteza Entorrinal/fisiología , Flavonoides/administración & dosificación , Guanilato Ciclasa/antagonistas & inhibidores , Hipocampo/efectos de los fármacos , Hipocampo/enzimología , Hipocampo/fisiología , Indoles/administración & dosificación , Inhibición Psicológica , Masculino , Memoria a Corto Plazo/efectos de los fármacos , Memoria a Corto Plazo/fisiología , Fenoles/administración & dosificación , Inhibidores de Proteínas Quinasas , Pirroles/administración & dosificación , Ratas , Ratas Wistar , Cloruro de Sodio/administración & dosificación , Estaurosporina/administración & dosificaciónRESUMEN
It is widely accepted that the formation of long-term memory (LTM) requires neuronal gene expression, protein synthesis and the remodeling of synaptic contacts. From mollusk to mammals, the cAMP/PKA/CREB signaling pathway has been shown to play a pivotal role in the establishment of LTM. More recently, the MAPK cascade has been also involved in memory processing. Here, we provide evidence for the participation of hippocampal PKA/CREB and MAPK/Elk-1 pathways, via activation of NMDA receptors, in memory formation of a one-trial avoidance learning in rats. Learning of this task is associated with an activation of p44 and p42 MAPKs, CREB and Elk-1, along with an increase in the levels of the catalytic subunit of PKA and Fos protein in nuclear-enriched hippocampal fractions. These changes were blocked by the immediate posttraining intra-hippocampal infusion of APV, a selective blocker of glutamate NMDA receptors, which renders the animals amnesic for this task. Moreover, no changes were found in control-shocked animals. Thus, inhibitory avoidance training in the rat is associated with an increase in the protein product of an IEG, c-fos, which occurs concomitantly with the activation of nuclear MAPK, CREB and Elk-1. NMDA receptors appear to be a necessary upstream step for the activation of these intracellular cascades during learning.
Asunto(s)
Reacción de Prevención , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Proteínas de Unión al ADN , Hipocampo/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Proteínas Proto-Oncogénicas/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Factores de Transcripción/metabolismo , 2-Amino-5-fosfonovalerato/farmacología , Animales , Electroforesis en Gel de Poliacrilamida , Antagonistas de Aminoácidos Excitadores/farmacología , Immunoblotting , Masculino , Microinyecciones , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Ratas , Ratas Wistar , Proteína Elk-1 con Dominio etsRESUMEN
6-Methyl-3'-bromoflavone inhibited [(3)H]flunitrazepam binding to the benzodiazepine binding site of the GABA(A) receptor (BDZ-bs) with Ki values between 10 and 50 nM in different brain regions. The GABA ratio of 1.03 for [(3)H]flunitrazepam binding to cerebral cortex, 0.76 for cerebellum, 0.7 for hippocampus, 0.7 for striatum, and 0.8 for spinal cord indicated an antagonistic or weak inverse agonistic profile of 6-methyl-3'-bromoflavone on BDZ-bs. Unlike classical benzodiazepines, it had no anticonvulsant, anxiolytic, myorelaxant, sedative, amnestic or motor incoordination effects. However, it antagonized the muscle relaxant, the sedative effect, and the changes in locomotor activity induced by diazepam. Taken together, these findings suggest that 6-methyl-3'-bromoflavone has an antagonistic profile on the BDZ-bs.
Asunto(s)
Encéfalo/metabolismo , Flavonoides/farmacología , Antagonistas del GABA/farmacología , Receptores de GABA-A/metabolismo , Médula Espinal/metabolismo , Sinaptosomas/metabolismo , Animales , Sitios de Unión , Unión Competitiva , Cerebelo/metabolismo , Corteza Cerebral/metabolismo , Cuerpo Estriado/metabolismo , Flavonoides/síntesis química , Flavonoides/química , Flunitrazepam/farmacocinética , Antagonistas de Receptores de GABA-A , Hipocampo/metabolismo , Membranas Intracelulares/metabolismo , Cinética , Ligandos , Masculino , RatonesRESUMEN
6-Bromo-3'-nitroflavone is a synthetic flavone derivative that selectively recognizes benzodiazepine receptors and has potent anxiolytic-like effects. Here, we describe in detail its pharmacological characterization. When i.p. injected in mice, 6-bromo-3'-nitroflavone (0.01-0.3 mg/kg) had an anxiolytic-like effect in the elevated plus-maze test. This effect was blocked by the specific benzodiazepine receptor antagonist, flumazenil. In addition, it exhibited anxiolytic-like actions when given orally (1 mg/kg). 6-Bromo-3'-nitroflavone did not exhibit myorelaxant effects (up to 30 mg/kg, i.p.). Unlike diazepam, this flavonoid produced no anterograde amnesia in a one-trial inhibitory avoidance learning. On the other hand, 6-bromo-3'-nitroflavone possessed mild anticonvulsant activity (0.1 mg/kg, i.p.) and provoked sedative-depressant actions only at doses 100-1000 times higher than those producing anxiolytic-like effects. 6-Bromo-3'-nitroflavone (0.1-1 mM) produced a lower potentiation of gamma-amino-butyric acid (GABA)-stimulated 36Cl- influx (126-138%) in comparison to diazepam (0.1 mM: 166%) in cerebral cortical membrane vesicles. Taken together, these findings suggest that 6-bromo-3'-nitroflavone has anxiolytic-like action possibly behaving as a partial agonist of the benzodiazepine receptors.
Asunto(s)
Ansiolíticos/farmacología , Flavonoides/farmacología , Agonistas de Receptores de GABA-A , Animales , Reacción de Prevención/efectos de los fármacos , Reacción de Prevención/fisiología , Flavonoides/uso terapéutico , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Ratones , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Pentobarbital/farmacología , Pentilenotetrazol/farmacología , Desempeño Psicomotor/efectos de los fármacos , Convulsiones/inducido químicamente , Convulsiones/tratamiento farmacológico , Sueño/efectos de los fármacos , Sueño/fisiologíaRESUMEN
Retrograde amnesia was induced in rats trained in step-down inhibitory avoidance by four different treatments: an ip injection of beta-endorphin (1.0 microgram kg), an electroconvulsive shock (ECS), an intrahippocampal infusion of the calcium/calmodulin protein kinase II inhibitor, KN62 (0.08 microgram/side), given 0 h after training, or an intrahippocampal infusion of the protein kinase A inhibitor, KT5720 (0.5 microgram/side), given 3 h after training. Pretest ip injections of ACTH (0.2 microgram/kg) or vasopressin (10.0 micrograms/kg), but not saline, reversed the amnesia caused by beta-endorphin and ECS but not that caused by the enzyme inhibitors. This suggests that the amnesia produced by intrahippocampal KN62 and KT5720 administration is stronger than that caused by ECS and beta-endorphin, possibly because the former interfere directly with specific steps of the core biochemical chain of events that underlies memory consolidation.
Asunto(s)
1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/análogos & derivados , Nivel de Alerta/efectos de los fármacos , Proteínas Quinasas Dependientes de Calcio-Calmodulina/antagonistas & inhibidores , Carbazoles , Cosintropina/farmacología , Inhibidores Enzimáticos/farmacología , Miedo/efectos de los fármacos , Hipocampo/efectos de los fármacos , Indoles/farmacología , Lipresina/farmacología , Recuerdo Mental/efectos de los fármacos , Pirroles/farmacología , betaendorfina/antagonistas & inhibidores , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina/farmacología , Animales , Nivel de Alerta/fisiología , Proteínas Quinasas Dependientes de Calcio-Calmodulina/fisiología , Relación Dosis-Respuesta a Droga , Miedo/fisiología , Hipocampo/fisiología , Inyecciones Intraperitoneales , Masculino , Vías Nerviosas/efectos de los fármacos , Vías Nerviosas/fisiología , Ratas , Ratas Wistar , Retención en Psicología/efectos de los fármacos , Retención en Psicología/fisiología , betaendorfina/farmacologíaRESUMEN
Male Wistar rats were trained in one-trial step-down inhibitory avoidance using a 0.4-mA footshock. At various times after training (0, 1.5, 3, 6 and 9 h for the animals implanted into the CA1 region of the hippocampus; 0 and 3 h for those implanted into the amygdala), these animals received microinfusions of SKF38393 (7.5 micrograms/side), SCH23390 (0.5 microgram/side), norepinephrine (0.3 microgram/side), timolol (0.3 microgram/side), 8-OH-DPAT (2.5 micrograms/side), NAN-190 (2.5 micrograms/side), forskolin (0.5 microgram/side), KT5720 (0.5 microgram/side) or 8-Br-cAMP (1.25 micrograms/side). Rats were tested for retention 24 h after training. When given into the hippocampus 0 h post-training, norepinephrine enhanced memory whereas KT5720 was amnestic. When given 1.5 h after training, all treatments were ineffective. When given 3 or 6 h post-training, 8-Br-cAMP, forskolin, SKF38393, norepinephrine and NAN-190 caused memory facilitation, while KT5720, SCH23390, timolol and 8-OH-DPAT caused retrograde amnesia. Again, at 9 h after training, all treatments were ineffective. When given into the amygdala, norepinephrine caused retrograde facilitation at 0 h after training. The other drugs infused into the amygdala did not cause any significant effect. These data suggest that in the hippocampus, but not in the amygdala, a cAMP/protein kinase A pathway is involved in memory consolidation at 3 and 6 h after training, which is regulated by D1, beta, and 5HT1A receptors. This correlates with data on increased post-training cAMP levels and a dual peak of protein kinase A activity and CREB-P levels (at 0 and 3-6 h) in rat hippocampus after training in this task. These results suggest that the hippocampus, but not the amygdala, is involved in long-term storage of step-down inhibitory avoidance in the rat.
Asunto(s)
Amígdala del Cerebelo/efectos de los fármacos , Proteínas Quinasas Dependientes de AMP Cíclico/efectos de los fármacos , AMP Cíclico/análisis , Hipocampo/efectos de los fármacos , Memoria/fisiología , 8-Bromo Monofosfato de Adenosina Cíclica/farmacología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Animales , Benzazepinas/farmacología , Colforsina/farmacología , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/análisis , Masculino , Norepinefrina/farmacología , Ratas , Ratas Wistar , Transducción de SeñalRESUMEN
Rats implanted bilaterally with cannulae in the CA1 region of the dorsal hippocampus or in the amygdala were trained in one-trial step-down inhibitory (passive) avoidance using a 0.4 mA footshock. At various times after training (0, 1.5, 3, 6 or 9 h for animals implanted in the hippocampus; 0 or 3 h for those implanted in the amygdala), they received infusions of 8-Br-cAMP (cyclic adenosine monophosphate) (1.25 micrograms/side), SKF38393 (7.5 micrograms/side), SCH23390 (0.5 microgram/side), norepinephrine ClH (0.3 microgram/side), timolol ClH (0.3 microgram/side), 8-HO-DPAT (2.5 micrograms/side), NAN-190 (2.5 micrograms/side), forskolin (0.5 microgram/side) or KT5720 (0.5 microgram/side). Rats were tested for retention 24 h after training. SKF38393 is an agonist and SCH23390 an antagonist at dopamine D1 receptors, timolol is a beta-adrenoceptor antagonist, 8-HO-DPAT is an agonist and NAN-190 an antagonist at 5HT1A receptors, forskolin enhances adenylyl cyclase, and KT5720 inhibits protein kinase A. When given into the hippocampus 0 h post-training, norepinephrine enhanced memory and KT5720 was amnestic. When given 1.5 h after training, all treatments were ineffective. When given 3 or 6 h post-training, 8-Br-cAMP, forskolin, SKF 38393, noradrenaline and NAN-190 caused memory facilitation, and KT5720, SCH23390, timolol and 8-HO-DPAT caused retrograde amnesia. At 9 h from training, all treatments were again ineffective. When given into the amygdala 0 or 3 h post-training all treatments were ineffective, except for noradrenaline at 0 h, which caused retrograde facilitation. The data agree with the suggestion that in the hippocampus, but not the amygdala, a cAMP/protein kinase A pathway is involved in memory consolidation at 3 and 6 h from training, and that this is regulated by D1, beta, and 5HT1A receptors. This correlates with a previous report of increased cAMP levels, protein kinase A activity and P-CREB levels at 3-6 h from training in rat hippocampus in this task. This may be taken to suggest that the hippocampus, but not the amygdala, is involved in the long-term storage of step-down inhibitory avoidance in the rat.
Asunto(s)
Carbazoles , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Memoria/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , 2,3,4,5-Tetrahidro-7,8-dihidroxi-1-fenil-1H-3-benzazepina/farmacología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos beta/farmacología , Amígdala del Cerebelo , Animales , Reacción de Prevención , Benzazepinas/farmacología , Colforsina/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/efectos de los fármacos , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Hipocampo , Indoles/farmacología , Masculino , Norepinefrina/farmacología , Piperazinas/farmacología , Pirroles/farmacología , Ratas , Ratas Wistar , Receptores Adrenérgicos beta/efectos de los fármacos , Receptores de Dopamina D1/efectos de los fármacos , Receptores de Serotonina/efectos de los fármacos , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Factores de Tiempo , Timolol/farmacologíaRESUMEN
Male Wistar rats were trained in one-trial step-down inhibitory avoidance using a 0.4-mA footshock. At various times after training (0, 1.5, 3,6 and 9 h for the animals implanted into the CA1 region of the hippocampus; 0 and 3 h for those implanted into the amygdala), these animals received microinfusions of SKF38393 (7.5 mug/side), SCH23390 (0.5 mug/side), norepinephrine (0.3 mug/side), timolol (0.3 mug/side), 8-OH-DPAT (2.5 mug/side), NAN-190 (2.5 mug/side), forskolin (0.5 mug/side), KT5720 (0.5 mug/side) or 8-Br-cAMP (1.25 mug/side). Rats were tested for retention 24 h after training. When given into the hippocampus 0 h post-training, norepinephrine enhanced memory whereas KT5720 was amnestic. When given 1.5 h after training, all treatments were ineffective. When given 3 or 6 h post-training, 8-Br-cAMP, forskolin, SKF38393, norepinephrine and NAN-190 caused memory facilitation, while KT5720, SCH23390, timolol and 8-OH-DPAT caused retrograde amnesia. Again, at 9 h after training, all treatments were inffective. When given into the amygdala, norepinephrine caused retrograde facilitation at 0 h after training. The other drugs infused into the amygdala did not cause any significant effect. These data suggest that in the hippocampus, but not in the amygdala, a cAMP/protein kinase A pathway is involved in memory cosolidation at 3 and 6 h after training, which is regulated by D1, Beta, and 5HT1A receptors. This correlates with data on increased post-training cAMP levels and a dual peak of protein kinase A activity and CREB-P levels (at 0 and 3-6 h) in rat hippocampus after training in this task. These results suggest that the hippocampus, but not the amygdala, is involved in long-term storage of step-down inhibitory avoidance in the rat.