RESUMEN
Presynaptic NMDA receptors facilitate the release of glutamate at excitatory cortical synapses and are involved in regulation of synaptic dynamics and plasticity. At synapses in the entorhinal cortex these receptors are tonically activated and provide a positive feedback modulation of the level of background excitation. NMDA receptor activation requires obligatory occupation of a co-agonist binding site, and in the present investigation we have examined whether this site on the presynaptic receptor is activated by endogenous glycine or d-serine. We used whole-cell patch clamp recordings of spontaneous AMPA receptor-mediated synaptic currents from rat entorhinal cortex neurones in vitro as a monitor of presynaptic glutamate release. Addition of exogenous glycine or d-serine had minimal effects on spontaneous release, suggesting that the co-agonist site was endogenously activated and likely to be saturated in our slices. This was supported by the observation that a co-agonist site antagonist reduced the frequency of spontaneous currents. Depletion of endogenous glycine by enzymatic breakdown with a bacterial glycine oxidase had little effect on glutamate release, whereas d-serine depletion with a yeast d-amino acid oxidase significantly reduced glutamate release, suggesting that d-serine is the endogenous agonist. Finally, the effects of d-serine depletion were mimicked by compromising astroglial cell function, and this was rescued by exogenous d-serine, indicating that astroglial cells are the provider of the d-serine that tonically activates the presynaptic NMDA receptor. We discuss the significance of these observations for the aetiology of epilepsy and possible targeting of the presynaptic NMDA receptor in anticonvulsant therapy.
Asunto(s)
Astrocitos/fisiología , Corteza Entorrinal/metabolismo , Terminales Presinápticos/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/agonistas , Receptores de N-Metil-D-Aspartato/metabolismo , Serina/farmacología , Animales , Corteza Entorrinal/efectos de los fármacos , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Ácido Glutámico/metabolismo , Glicina/farmacología , Ratas , Ratas WistarRESUMEN
Ethosuximide is the drug of choice for treating generalized absence seizures, but its mechanism of action is still a matter of debate. It has long been thought to act by disrupting a thalamic focus via blockade of T-type channels and, thus, generation of spike-wave activity in thalamocortical pathways. However, there is now good evidence that generalized absence seizures may be initiated at a cortical focus and that ethosuximide may target this focus. In the present study we have looked at the effect ethosuximide on glutamate and GABA release at synapses in the rat entorhinal cortex in vitro, using two experimental approaches. Whole-cell patch-clamp studies revealed an increase in spontaneous GABA release by ethosuximide concurrent with no change in glutamate release. This was reflected in studies that estimated global background inhibition and excitation from intracellularly recorded membrane potential fluctuations, where there was a substantial rise in the ratio of network inhibition to excitation, and a concurrent decrease in excitability of neurones embedded in this network. These studies suggest that, in addition to well-characterised effects on ion channels, ethosuximide may directly elevate synaptic inhibition in the cortex and that this could contribute to its anti-absence effects. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.
Asunto(s)
Anticonvulsivantes/farmacología , Corteza Entorrinal/efectos de los fármacos , Etosuximida/farmacología , Red Nerviosa/efectos de los fármacos , Neuronas/efectos de los fármacos , Ácido gamma-Aminobutírico/metabolismo , Animales , Corteza Entorrinal/fisiología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Masculino , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Red Nerviosa/fisiología , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Neuronas/fisiología , Técnicas de Placa-Clamp , Ratas , Ratas Wistar , Sinapsis/efectos de los fármacos , Sinapsis/fisiología , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiologíaRESUMEN
We applied the group-I metabotropic glutamate (mGlu) receptor agonist, 3,5-dihydroxyphenylglycine (DHPG), to neonatal or adult rat hippocampal slices at concentrations (10 microM) that induced a short-term depression (STD) of excitatory synaptic transmission at the Schaffer collateral/CA1 synapses. DHPG-induced STD was entirely mediated by the activation of mGlu5 receptors because it was abrogated by the mGlu5 receptor antagonist, MPEP [2-methyl-6-(phenylethynyl)pyridine], but not by the mGlu1 receptor antagonist, CPCCOEt [7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester]. Knowing that ephrin-Bs functionally interact with group-I mGlu receptors (Calò et al., 2005), we examined whether pharmacological activation of ephrin-Bs could affect DHPG-induced STD. We activated ephrin-Bs using their cognate receptor, EphB1, under the form of a preclustered EphB1/Fc chimera. Addition of clustered EphB1/Fc alone to the slices induced a small but nondecremental depression of excitatory synaptic transmission, which differed from the depression induced by 10 microM DHPG. Surprisingly, EphB1/Fc-induced synaptic depression was abolished by MPEP (but not by CPCCOEt) suggesting that it required the endogenous activation of mGlu5 receptors. In addition, coapplication of DHPG and EphB1/Fc, resulted in a large and nondecremental long-term depression. The effect of clustered EphB1/Fc was specific because it was not mimicked by unclustered EphB1/Fc or clustered EphA1/Fc. These findings raise the intriguing possibility that changes in synaptic efficacy mediated by mGlu5 receptors are under the control of the ephrin/Eph receptor system, and that the neuronal actions of ephrins can be targeted by drugs that attenuate mGlu5 receptor signaling.
Asunto(s)
Efrinas/metabolismo , Ácido Glutámico/metabolismo , Hipocampo/metabolismo , Depresión Sináptica a Largo Plazo/fisiología , Receptores de Glutamato Metabotrópico/metabolismo , Transmisión Sináptica/fisiología , Animales , Efrina-B1/agonistas , Efrina-B1/metabolismo , Agonistas de Aminoácidos Excitadores/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/citología , Masculino , Técnicas de Cultivo de Órganos , Técnicas de Placa-Clamp , Piridinas/farmacología , Ratas , Ratas Wistar , Receptor EphB1/genética , Receptor EphB1/metabolismo , Receptor del Glutamato Metabotropico 5 , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Transmisión Sináptica/efectos de los fármacosRESUMEN
The perirhinal cortex of the temporal lobe is essential for the familiarity discrimination component of recognition memory. In view of the importance of changes in calcium ion concentration for synaptic plasticity, the present study examined the effects of L-type voltage-dependent calcium channel (VDCC) antagonism on rat perirhinal-based familiarity discrimination processes and plasticity including long-term depression (LTD), long-term potentiation (LTP), and depotentiation. Single doses of three different types of L-type VDCC antagonists, verapamil, diltiazem, and nifedipine, administered systemically, or verapamil administered locally into the perirhinal cortex, impaired acquisition of long-term (24 h) but not shorter-term (20 min) recognition memory. L-type VDCC antagonism also disrupted memory retrieval after 24 h but not 20 min. Differential neuronal activation produced by viewing novel or familiar visual stimuli was measured by Fos expression. L-type VDCC antagonism by verapamil in perirhinal cortex during memory acquisition disrupted the normal pattern of differential Fos expression, so paralleling the antagonist-induced memory impairment. In slices of perirhinal cortex maintained in vitro, verapamil was without effect on baseline excitability or LTP but blocked LTD and depotentiation. The consistency of effects across the behavioral and cellular levels of analysis provides strong evidence for the involvement of perirhinal L-type VDCCs in long-term recognition memory processes.
Asunto(s)
Canales de Calcio Tipo L/metabolismo , Plasticidad Neuronal/fisiología , Reconocimiento en Psicología/fisiología , Lóbulo Temporal/fisiología , Animales , Bloqueadores de los Canales de Calcio/administración & dosificación , Diltiazem/administración & dosificación , Técnicas In Vitro , Potenciación a Largo Plazo/efectos de los fármacos , Potenciación a Largo Plazo/fisiología , Depresión Sináptica a Largo Plazo/efectos de los fármacos , Depresión Sináptica a Largo Plazo/fisiología , Plasticidad Neuronal/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/fisiología , Nifedipino/administración & dosificación , Proteínas Proto-Oncogénicas c-fos/metabolismo , Ratas , Reconocimiento en Psicología/efectos de los fármacos , Sinapsis/efectos de los fármacos , Sinapsis/fisiología , Lóbulo Temporal/efectos de los fármacos , Factores de Tiempo , Verapamilo/administración & dosificaciónRESUMEN
Recent, convergent evidence places the anterior thalamic nuclei at the heart of diencephalic amnesia. However, the reasons for the severe memory loss in diencephalic amnesia remain unknown. A potential clue comes from the dense, reciprocal connections between the anterior thalamic nuclei and retrosplenial cortex, another region vital for memory. We now report a loss of synaptic plasticity [long-term depression (LTD)] in rat retrosplenial cortex slices months following an anterior thalamic lesion. The loss of LTD was lamina-specific, occurring only in superficial layers of the cortex and was associated with a decrease in GABA(A)-mediated inhibitory transmission. As retrosplenial cortex is itself vital for memory, this distal lesion effect will amplify the impact of anterior thalamic lesions. These findings not only provide novel insights into the functional pathology of diencephalic amnesia and have implications for the aetiology of the posterior cingulate hypoactivity in Alzheimer's disease, but also show how distal changes in plasticity could contribute to diaschisis.
Asunto(s)
Amnesia/patología , Núcleos Talámicos Anteriores/patología , Giro del Cíngulo/patología , Plasticidad Neuronal , Sinapsis/fisiología , Amnesia/metabolismo , Amnesia/fisiopatología , Animales , Núcleos Talámicos Anteriores/metabolismo , Núcleos Talámicos Anteriores/fisiopatología , Giro del Cíngulo/fisiopatología , Masculino , N-Metilaspartato/metabolismo , Técnicas de Placa-Clamp , Ratas , Receptores de N-Metil-D-Aspartato/fisiología , Transmisión Sináptica/fisiología , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Learning is widely believed to involve synaptic plasticity, using mechanisms such as those used in long-term potentiation (LTP). We assess whether the mechanisms used in alternative forms of plasticity, long-term depression (LTD) and depotentiation, play a role in learning. We have exploited the involvement of the perirhinal cortex in two different forms of learning to compare simultaneously, within the same brain region, their effects on LTD and depotentiation. Multiple-exposure learning but not single-exposure learning in vivo prevented, in a muscarinic receptor-dependent manner, subsequent induction of LTD and depotentiation, but not LTP, in perirhinal cortex in vitro. The contrast in the effects of the two types of learning under these particular experimental conditions indicate that the in vitro change is unlikely to be attributable to synapse-specific plastic changes registering the precise details of the individual learned associations. Instead, it is concluded that the lack of LTD and depotentiation arises from, and establishes the importance of, a learning-related generalized change in plasticity gain. The existence of this additional mechanism has important implications for interpretations of how plasticity relates to learning.
Asunto(s)
Corteza Cerebral/fisiología , Aprendizaje/fisiología , Depresión Sináptica a Largo Plazo/fisiología , Animales , Conducta Animal , Relación Dosis-Respuesta en la Radiación , Estimulación Eléctrica/métodos , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/efectos de la radiación , Técnicas In Vitro , Masculino , Antagonistas Muscarínicos/farmacología , Estimulación Luminosa/métodos , Ratas , Reconocimiento en Psicología/fisiología , Escopolamina/farmacologíaRESUMEN
Long-term potentiation (LTP) and long-term depression (LTD) remain widely accepted vertebrate models for the cellular and molecular mechanisms that underlie synaptic changes during learning and memory. Although LTD is a phenomenon that occurs in many regions of the CNS, it is clear that the mechanisms recruited in its induction and expression can vary, depending on many factors, including brain region and developmental time point. LTD in the hippocampus and cerebellum is probably the best characterized, although there are also other brain areas where mechanisms of LTD are well understood, and where it is thought to have a functional role.
Asunto(s)
Encéfalo/fisiología , Depresión Sináptica a Largo Plazo/fisiología , Transmisión Sináptica/fisiología , Animales , Encéfalo/anatomía & histología , Humanos , Modelos Neurológicos , Red Nerviosa/anatomía & histología , Red Nerviosa/fisiologíaRESUMEN
The role of NMDA receptors in the induction of long-term potentiation (LTP) and long-term depression (LTD) is well established but which particular NR2 subunits are involved in these plasticity processes is still a matter of controversy. We have studied the effects of subtype selective NMDA receptor antagonists on LTP induced by high frequency stimulation (100 Hz for 1s) and LTD induced by low frequency stimulation (1 Hz for 15 min) in the CA1 region of hippocampal slices from 14 day old Wistar rats. Against recombinant receptors in HEK293 cells NVP-AAM077 (NVP) was approximately 14-fold selective for NR2A vs NR2B receptors, whilst Ro 25-6981 (Ro) was highly selective for NR2B receptors. On NMDA receptor-mediated EPSCs from Schaffer collaterals in CA1 neurones, NVP and Ro both reduced the amplitude but differentially affected the time constant of decay. The data are compatible with the selective effect of NVP (0.1 microM) and Ro (4 microM) on native NR2A and NBR2B receptors, respectively. NVP reduced both LTP and LTD whereas Ro reduced only LTP. Thus, LTP was reduced by 63% at 0.1 microM NVP and almost completely at 0.4 microM whereas 5 microM Ro reduced LTP by 45%. These data are consistent with a role for both NR2A and NR2B in the induction of LTP, under our experimental conditions. In comparison, LTD was unaffected by Ro (5 microM) even in the presence of a glutamate uptake inhibitor threo-beta-benzylaspartic acid (TBOA) to increase the concentration of glutamate at NR2B containing receptors. NVP (0.2-0.4 microM), however, produced a concentration dependent inhibition of LTD which was complete at 0.4 microM. The lack of effect of 0.1 microM NVP on LTD contrasts with its marked effect on LTP and raises the possibility that different NVP-sensitive NR2 subunit-containing NMDA receptors are required for LTP and LTD in this preparation.
Asunto(s)
Hipocampo/fisiología , Potenciación a Largo Plazo/fisiología , Depresión Sináptica a Largo Plazo/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Animales , Animales Recién Nacidos , Línea Celular Transformada , Relación Dosis-Respuesta a Droga , Relación Dosis-Respuesta en la Radiación , Interacciones Farmacológicas , Estimulación Eléctrica/métodos , Agonistas de Aminoácidos Excitadores/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Femenino , Humanos , Técnicas In Vitro , Potenciación a Largo Plazo/efectos de los fármacos , Potenciación a Largo Plazo/efectos de la radiación , Depresión Sináptica a Largo Plazo/efectos de los fármacos , Depresión Sináptica a Largo Plazo/efectos de la radiación , N-Metilaspartato/farmacología , Técnicas de Placa-Clamp/métodos , Fenoles/farmacología , Piperidinas/farmacología , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Transfección/métodosRESUMEN
Mechanisms of long-term potentiation and depression (LTP and LTD) change considerably during development, but the importance of these changes and the factors that control them is not clear. We found that visual experience triggered a switch in mechanisms of LTD in rat perirhinal cortex, an area critical for visual recognition memory. Thus, changes in synaptic plasticity mechanisms were correlated with the changing physiological demands on the CNS.
Asunto(s)
Corteza Cerebral/crecimiento & desarrollo , Depresión Sináptica a Largo Plazo/fisiología , Plasticidad Neuronal/fisiología , Animales , Corteza Cerebral/metabolismo , Luz , Técnicas de Cultivo de Órganos , Ratas , Receptores de Glutamato/biosíntesis , Receptores Muscarínicos/biosíntesis , Visión Ocular/fisiologíaRESUMEN
We established the importance of phosphorylation of cAMP responsive element-binding protein (CREB) to both the familiarity discrimination component of long-term recognition memory and plasticity within the perirhinal cortex of the temporal lobe. Adenoviral transduction of perirhinal cortex (and adjacent visual association cortex) with a dominant-negative inhibitor of CREB impaired the preferential exploration of novel over familiar objects at a long (24 h) but not a short (15 min) delay, disrupted the normal reduced activation of perirhinal neurons to familiar compared with novel pictures, and impaired long-term potentiation of synaptic transmission in perirhinal slices. The consistency of these effects across the behavioral, systems, and cellular levels of analysis provides strong evidence for involvement of CREB phosphorylation in synaptic plastic processes within perirhinal cortex necessary for long-term recognition memory.
Asunto(s)
Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Conducta Exploratoria/fisiología , Potenciación a Largo Plazo/fisiología , Reconocimiento Visual de Modelos/fisiología , Procesamiento Proteico-Postraduccional , Lóbulo Temporal/fisiología , Adenoviridae/genética , Animales , Aprendizaje por Asociación , AMP Cíclico/fisiología , Virus Defectuosos/genética , Aprendizaje Discriminativo/fisiología , Discriminación en Psicología/fisiología , Corteza Entorrinal/fisiología , Potenciales Postsinápticos Excitadores , Genes Reporteros , Genes fos , Vectores Genéticos/genética , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/genética , Técnicas In Vitro , Masculino , Fosforilación , Estimulación Luminosa , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Ratas , Proteínas Recombinantes de Fusión/genética , Método Simple Ciego , Transmisión Sináptica/fisiología , Transcripción Genética , Transducción GenéticaRESUMEN
It is widely believed that long-term depression (LTD) and its counterpart, long-term potentiation (LTP), involve mechanisms that are crucial for learning and memory. However, LTD is difficult to induce in adult cortex for reasons that are not known. Here we show that LTD can be readily induced in adult cortex by the activation of NMDA receptors (NMDARs), after inhibition of glutamate uptake. Interestingly there is no need to activate synaptic NMDARs to induce this LTD, suggesting that LTD is triggered primarily by extrasynaptic NMDA receptors. We also find that de novo LTD requires the activation of NR2B-containing NMDAR, whereas LTP requires activation of NR2A-containing NMDARs. Surprisingly another form of LTD, depotentiation, requires activation of NR2A-containing NMDARs. Therefore, NMDARs with different synaptic locations and subunit compositions are involved in various forms of synaptic plasticity in adult cortex.
Asunto(s)
Corteza Cerebral/fisiología , Potenciación a Largo Plazo/fisiología , Depresión Sináptica a Largo Plazo/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , 2-Amino-5-fosfonovalerato/farmacología , Animales , Ácido Aspártico/farmacología , Corteza Cerebral/efectos de los fármacos , Ácidos Dicarboxílicos/farmacología , Maleato de Dizocilpina/farmacología , Agonistas de Aminoácidos Excitadores/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Ácido Glutámico/farmacología , Potenciación a Largo Plazo/efectos de los fármacos , Depresión Sináptica a Largo Plazo/efectos de los fármacos , N-Metilaspartato/farmacología , Neuronas/efectos de los fármacos , Neuronas/fisiología , Neuronas/ultraestructura , Inhibidores de la Captación de Neurotransmisores/farmacología , Fenoles/farmacología , Picrotoxina/farmacología , Piperidinas/farmacología , Subunidades de Proteína , Pirrolidinas/farmacología , Quinoxalinas/farmacología , Ratas , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/efectos de los fármacosRESUMEN
We establish the importance of cholinergic neurotransmission to both recognition memory and plasticity within the perirhinal cortex of the temporal lobe. The muscarinic receptor antagonist scopolamine impaired the preferential exploration of novel over familiar objects, disrupted the normal reduced activation of perirhinal neurones to familiar compared to novel pictures, and blocked production of long-term depression (LTD) but not long-term potentiation (LTP) of synaptic transmission in perirhinal slices. The consistency of these effects across the behavioral, systems, and cellular levels of analysis provides strong evidence for the involvement of cholinergic mechanisms in synaptic plastic processes within perirhinal cortex that are necessary for recognition memory.