RESUMEN
The triple-network model of psychopathology is a framework to explain the functional and structural neuroimaging phenotypes of psychiatric and neurological disorders. It describes the interactions within and between three distributed networks: the salience, default-mode, and central executive networks. These have been associated with brain disorder traits in patients. Homologous networks have been proposed in animal models, but their integration into a triple-network organization has not yet been determined. Using resting-state datasets, we demonstrate conserved spatio-temporal properties between triple-network elements in human, macaque, and mouse. The model predictions were also shown to apply in a mouse model for depression. To validate spatial homologies, we developed a data-driven approach to convert mouse brain maps into human standard coordinates. Finally, using high-resolution viral tracers in the mouse, we refined an anatomical model for these networks and validated this using optogenetics in mice and tractography in humans. Unexpectedly, we find serotonin involvement within the salience rather than the default-mode network. Our results support the existence of a triple-network system in the mouse that shares properties with that of humans along several dimensions, including a disease condition. Finally, we demonstrate a method to humanize mouse brain networks that opens doors to fully data-driven trans-species comparisons.
Asunto(s)
Imagen por Resonancia Magnética , Red Nerviosa , Animales , Encéfalo , Mapeo Encefálico/métodos , Humanos , Imagen por Resonancia Magnética/métodos , Ratones , Vías NerviosasRESUMEN
This study proposes a new intelligent diagnostic method for bearing faults in rotating machinery. The method uses a combination of nonlinear mode decomposition based on the improved fast kurtogram, gramian angular field, and convolutional neural network to detect the bearing state of rotating machinery. The nonlinear mode decomposition based on the improved fast kurtogram inherits the advantages of the original algorithm while improving the computational efficiency and signal-to-noise ratio. The gramian angular field can construct a two-dimensional image without destroying the time relationship of the signal. Therefore, the proposed method can perform fault diagnosis on rotating machinery under complex operating conditions. The proposed method is verified on the Paderborn dataset under heavy noise and multiple operating conditions to evaluate its effectiveness. Experimental results show that the proposed model outperforms wavelet denoising and the traditional adaptive decomposition method. The proposed model achieves over 99.6% accuracy in all four operating conditions provided by this dataset, and 93.8% accuracy in a strong noise environment with a signal-to-noise ratio of -4 dB.
Asunto(s)
Análisis de Falla de Equipo , Redes Neurales de la Computación , Algoritmos , Análisis de Falla de Equipo/métodos , Humanos , Reproducibilidad de los Resultados , Relación Señal-RuidoRESUMEN
Fast exchange of extracellular signals between neurons and astrocytes is crucial for synaptic function. Over the last few decades, different pathways of astroglial release of neuroactive substances have been proposed to modulate neurotransmission. However, their involvement in physiological conditions is highly debated. Connexins, the gap junction forming proteins, are highly expressed in astrocytes and have recently been shown to scale synaptic transmission and plasticity. Interestingly, in addition to gap junction channels, the most abundant connexin (Cx) in astrocytes, Cx43, also forms hemichannels. While such channels are mostly active in pathological conditions, they have recently been shown to regulate cognitive function. However, whether astroglial Cx43 hemichannels are active in resting conditions and regulate basal synaptic transmission is unknown. Here we show that in basal conditions Cx43 forms functional hemichannels in astrocytes from mouse hippocampal slices. We furthermore demonstrate that the activity of astroglial Cx43 hemichannels in resting states regulates basal excitatory synaptic transmission of hippocampal CA1 pyramidal cells through ATP signaling. These data reveal Cx43 hemichannels as a novel astroglial release pathway at play in basal conditions, which tunes the moment-to-moment glutamatergic synaptic transmission.
Asunto(s)
Astrocitos/metabolismo , Conexina 43/metabolismo , Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/citología , Transmisión Sináptica/fisiología , Adenosina Trifosfato/metabolismo , Adenosina Trifosfato/farmacología , Animales , Animales Recién Nacidos , Astrocitos/efectos de los fármacos , Carbenoxolona/farmacología , Conexina 43/genética , Estimulación Eléctrica , Etidio/metabolismo , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Técnicas In Vitro , Ratones , Ratones Transgénicos , Neuronas/efectos de los fármacos , Neuronas/fisiología , Técnicas de Placa-Clamp , Péptidos/farmacología , Antagonistas del Receptor Purinérgico P2/farmacología , Fosfato de Piridoxal/análogos & derivados , Fosfato de Piridoxal/farmacología , Transmisión Sináptica/efectos de los fármacosRESUMEN
Functional network activity alterations are one of the earliest hallmarks of Alzheimer's disease (AD), detected prior to amyloidosis and tauopathy. Better understanding the neuronal underpinnings of such network alterations could offer mechanistic insight into AD progression. Here, we examined a mouse model (3xTgAD mice) recapitulating this early AD stage. We found resting functional connectivity loss within ventral networks, including the entorhinal cortex, aligning with the spatial distribution of tauopathy reported in humans. Unexpectedly, in contrast to decreased connectivity at rest, 3xTgAD mice show enhanced fMRI signal within several projection areas following optogenetic activation of the entorhinal cortex. We corroborate this finding by demonstrating neuronal facilitation within ventral networks and synaptic hyperexcitability in projection targets. 3xTgAD mice, thus, reveal a dichotomic hypo-connected:resting versus hyper-responsive:active phenotype. This strong homotopy between the areas affected supports the translatability of this pathophysiological model to tau-related, early-AD deficits in humans.
Asunto(s)
Enfermedad de Alzheimer , Tauopatías , Enfermedad de Alzheimer/metabolismo , Animales , Modelos Animales de Enfermedad , Corteza Entorrinal , Humanos , Ratones , Ratones Transgénicos , Neuronas/metabolismo , Tauopatías/diagnóstico por imagen , Tauopatías/metabolismo , Proteínas tau/genética , Proteínas tau/metabolismoRESUMEN
The motivation to eat is not only shaped by nutrition but also competed by external stimuli including pain. How the mouse hypothalamus, the feeding regulation center, integrates nociceptive inputs to modulate feeding is unclear. Within the key nociception relay center parabrachial nucleus (PBN), we demonstrated that neurons projecting to the lateral hypothalamus (LHPBN) are nociceptive yet distinct from danger-encoding central amygdala-projecting (CeAPBN) neurons. Activation of LHPBN strongly suppressed feeding by limiting eating frequency and also reduced motivation to work for food reward. Refined approach-avoidance paradigm revealed that suppression of LHPBN, but not CeAPBN, sustained motivation to obtain food. The effect of LHPBN neurons on feeding was reversed by suppressing downstream LHVGluT2 neurons. Thus, distinct from a circuit for fear and escape responses, LHPBN neurons channel nociceptive signals to LHVGluT2 neurons to suppress motivational drive for feeding. Our study provides a new perspective in understanding feeding regulation by external competing stimuli.
RESUMEN
Despite notable genetic influences, obesity mainly results from the overconsumption of food, which arises from the interplay of physiological, cognitive and environmental factors. In patients with obesity, eating is determined more by external cues than by internal physiological needs. However, how environmental context drives non-homeostatic feeding is elusive. Here, we identify a population of somatostatin (TNSST) neurons in the mouse hypothalamic tuberal nucleus that are preferentially activated by palatable food. Activation of TNSST neurons enabled a context to drive non-homeostatic feeding in sated mice and required inputs from the subiculum. Pairing a context with palatable food greatly potentiated synaptic transmission between the subiculum and TNSST neurons and drove non-homeostatic feeding that could be selectively suppressed by inhibiting TNSST neurons or the subiculum but not other major orexigenic neurons. These results reveal how palatable food, through a specific hypothalamic circuit, empowers environmental context to drive non-homeostatic feeding.
Asunto(s)
Conducta Alimentaria/fisiología , Hipotálamo/fisiología , Vías Nerviosas/fisiología , Neuronas/fisiología , Animales , Señales (Psicología) , Masculino , Ratones , Somatostatina/metabolismoRESUMEN
The tuberal nucleus (TN) is a surprisingly understudied brain region. We found that somatostatin (SST) neurons in the TN, which is known to exhibit pathological or cytological changes in human neurodegenerative diseases, play a crucial role in regulating feeding in mice. GABAergic tuberal SST (TNSST) neurons were activated by hunger and by the hunger hormone, ghrelin. Activation of TNSST neurons promoted feeding, whereas inhibition reduced it via projections to the paraventricular nucleus and bed nucleus of the stria terminalis. Ablation of TNSST neurons reduced body weight gain and food intake. These findings reveal a previously unknown mechanism of feeding regulation that operates through orexigenic TNSST neurons, providing a new perspective for understanding appetite changes.
Asunto(s)
Regulación del Apetito/fisiología , Neuronas GABAérgicas/fisiología , Somatostatina/fisiología , Núcleos Talámicos Ventrales/fisiología , Animales , Ghrelina/fisiología , Ratones , Ratones Mutantes , Núcleo Hipotalámico Paraventricular/citología , Núcleo Hipotalámico Paraventricular/fisiología , Núcleos Talámicos Ventrales/citologíaRESUMEN
The main energy source powering the brain is glucose. Strong energy needs of our nervous system are fulfilled by conveying this essential metabolite through blood via an extensive vascular network. Glucose then reaches brain tissues by cell uptake, diffusion and metabolization, processes primarily undertaken by astrocytes. Deprivation of glucose can however occur in various circumstances. In particular, ageing is associated with cognitive disturbances that are partly attributable to metabolic deficiency leading to brain glycopenia. Despite the crucial role of glucose and its metabolites in sustaining neuronal activity, little is known about its moment-to-moment contribution to astroglial physiology. We thus here investigated the early structural and functional alterations induced in astrocytes by a transient metabolic challenge consisting in glucose deprivation. Electrophysiological recordings of hippocampal astroglial cells of the stratum radiatum in situ revealed that shortage of glucose specifically increases astrocyte membrane capacitance, whilst it has no impact on other passive membrane properties. Consistent with this change, morphometric analysis unraveled a prompt increase in astrocyte volume upon glucose deprivation. Furthermore, characteristic functional properties of astrocytes are also affected by transient glucose deficiency. We indeed found that glucoprivation decreases their gap junction-mediated coupling, while it progressively and reversibly increases their intracellular calcium levels during the slow depression of synaptic transmission occurring simultaneously, as assessed by dual electrophysiological and calcium imaging recordings. Together, these data indicate that astrocytes rapidly respond to metabolic dysfunctions, and are therefore central to the neuroglial dialog at play in brain adaptation to glycopenia.
RESUMEN
The secreted leucine-rich glioma inactivated 1 (LGI1) protein is an important actor for human seizures of both genetic and autoimmune etiology: mutations in LGI1 cause inherited temporal lobe epilepsy, while LGI1 is involved in antibody-mediated encephalitis. Remarkably, Lgi1-deficient (Lgi1(-/-)) mice recapitulate the epileptic disorder and display early-onset spontaneous seizures. To understand how Lgi1-deficiency leads to seizures during postnatal development, we here investigated the early functional and structural defects occurring before seizure onset in Lgi1(-/-) mice. We found an increased excitatory synaptic transmission in hippocampal slices from Lgi1(-/-) mice. No structural alteration in the morphology of pyramidal cell dendrites and synapses was observed at this stage, indicating that Lgi1-deficiency is unlikely to trigger early developmental abnormalities. Consistent with the presynaptic subcellular localization of the protein, Lgi1-deficiency caused presynaptic defects, with no alteration in postsynaptic AMPA receptor activity in Lgi1-/- pyramidal cells before seizure onset. Presynaptic dysfunction led to increased synaptic glutamate levels, which were associated with hyperexcitable neuronal networks. Altogether, these data show that Lgi1 acts presynaptically as a negative modulator of excitatory synaptic transmission during early postnatal development. We therefore here reveal that increased presynaptic glutamate release is a key early event resulting from Lgi1-deficiency, which likely contributes to epileptogenesis.
Asunto(s)
Epilepsia/fisiopatología , Regulación del Desarrollo de la Expresión Génica , Proteínas/metabolismo , Transmisión Sináptica , Animales , Modelos Animales de Enfermedad , Femenino , Hipocampo/fisiopatología , Péptidos y Proteínas de Señalización Intracelular , Masculino , Ratones Noqueados , Proteínas/genética , Células Piramidales/fisiologíaRESUMEN
Neuronal networks can generate complex patterns of activity that depend on membrane properties of individual neurons as well as on functional synapses. To decipher the impact of synaptic properties and connectivity on neuronal network behavior, we investigate the responses of neuronal ensembles from small (5-30 cells in a restricted sphere) and large (acute hippocampal slice) networks to single electrical stimulation: in both cases, a single stimulus generated a synchronous long-lasting bursting activity. While an initial spike triggered a reverberating network activity that lasted 2-5 seconds for small networks, we found here that it lasted only up to 300 milliseconds in slices. To explain this phenomena present at different scales, we generalize the depression-facilitation model and extracted the network time constants. The model predicts that the reverberation time has a bell shaped relation with the synaptic density, revealing that the bursting time cannot exceed a maximum value. Furthermore, before reaching its maximum, the reverberation time increases sub-linearly with the synaptic density of the network. We conclude that synaptic dynamics and connectivity shape the mean burst duration, a property present at various scales of the networks. Thus bursting reverberation is a property of sufficiently connected neural networks, and can be generated by collective depression and facilitation of underlying functional synapses.
Asunto(s)
Hipocampo/metabolismo , Neuronas/metabolismo , Sinapsis/fisiología , Estimulación Eléctrica , Humanos , Depresión Sináptica a Largo Plazo/fisiología , Modelos TeóricosRESUMEN
Phenytoin is an effective anti-epileptic drug that inhibits Na(+) channel activities; however, how phenytoin modulates synaptic transmission to soothe epileptic symptoms is not clear. To characterize the effects of phenytoin regulation on neurotransmission, we studied the electrophysical properties of cultured embryonic cortical neurons. Phenytoin inhibited the inward Na(+) current in a dose-dependent manner with an IC50 of 16.8 µM, and at 100 µM, the inhibitory effect of phenytoin on the Na(+) current was proportional to the frequency applied. In cultured neurons, phenytoin significantly decreased the action potential firing rate and the peak potential. To study the effect of phenytoin in neurotransmission, we measured the Ca(2+) responses from stimulated target neurons and their neighboring neurons. Phenytoin significantly suppressed the Ca(2+) responses evoked by strong stimulations in the target and neighboring neurons, and exerted a decreased inhibitory effect under moderate stimulation. Picrotoxin, a GABAA receptor antagonist, enhanced the recorded spontaneous excitatory postsynaptic current activities. After picrotoxin-induced enhancement, phenytoin had a more pronounced effect on the suppression of the spontaneous hyper-exciting excitatory postsynaptic current (>100 pA), but it only mildly inhibited the general excitatory postsynaptic current. Our results demonstrate that phenytoin suppresses the efficacy of neurotransmission especially for the high-frequency stimulation by reducing the Na(+) channel activity and can potentially alleviate epileptiform activity.
Asunto(s)
Anticonvulsivantes/farmacología , Corteza Cerebral/efectos de los fármacos , Neuronas/efectos de los fármacos , Fenitoína/farmacología , Transmisión Sináptica/efectos de los fármacos , Animales , Células Cultivadas , Corteza Cerebral/embriología , Potenciales de la Membrana/efectos de los fármacos , Ratas Sprague-DawleyRESUMEN
γ-Aminobutyric acid (GABA) spillover from synaptic cleft activates extrasynaptic GABAA receptor and results in a tonic inhibition, which induces a background inhibitory effect to stabilize the membrane potential of the neuronal cells. However, the role of tonic inhibition and how it can be regulated during brain development and epileptic state remain elusive. By whole-cell patch-clamp recording on the granule cell in the dentate gyrus, we recorded tonic conductance to investigate the level of tonic inhibition in these two critical periods. According to our observation, an age-dependent increase in tonic conductance was observed. Furthermore, a change in tonic inhibition was also found in a chronic epileptic animal model, indicating that the alteration in tonic inhibition after epilepsy induction persists for a long duration to modulate neuronal activities. The present results show that tonic inhibition is altered during brain development and a chronic epileptic condition, indicating a role of the tonic inhibitory effect in both the critical periods.
Asunto(s)
Envejecimiento/fisiología , Giro Dentado/fisiopatología , Epilepsia/fisiopatología , Ácido gamma-Aminobutírico/fisiología , Animales , Anticonvulsivantes/farmacología , Interpretación Estadística de Datos , Diazepam/farmacología , Fenómenos Electrofisiológicos , Agonistas del GABA/farmacología , Moduladores del GABA/farmacología , Técnicas In Vitro , Masculino , Ácidos Nipecóticos/farmacología , Técnicas de Placa-Clamp , Ratas , Ratas Wistar , Estado Epiléptico/fisiopatología , TiagabinaRESUMEN
Synaptic transmission depends on several molecular and geometric components, such as the location of vesicular release, the number of released neurotransmitter molecules, the number and type of receptors, as well as the synapse organization. Our goal here is to illustrate how synaptic modeling allows extracting quantitative information in the context of neurological diseases and associated therapies. Combining electrophysiology with simulation tools, we first evaluate the reduction in synaptically released glutamate molecules induced by a ketogenic diet. In a second part, because the scaffolding molecule Shank3 is disrupted at the postsynaptic density in Autism Spectral Disorders, we present a numerical simulation of the synaptic response where this disruption leads to an alteration of the excitatory AMPA receptor trafficking. The take home message is that combining recent experimental findings with modeling approaches allows obtaining precise quantitative properties of what was still unapproachable a decade ago.
RESUMEN
This study was undertaken to assay the effect of lovastatin on the glycogen synthase kinase-3 beta (GSK-3ß) and collapsin responsive mediator protein-2 (CRMP-2) signaling pathway and mossy fiber sprouting (MFS) in epileptic rats. MFS in the dentate gyrus (DG) is an important feature of temporal lobe epilepsy (TLE) and is highly related to the severity and the frequency of spontaneous recurrent seizures. However, the molecular mechanism of MFS is mostly unknown. GSK-3ß and CRMP-2 are the genes responsible for axonal growth and neuronal polarity in the hippocampus, therefore this pathway is a potential target to investigate MFS. Pilocarpine-induced status epilepticus animal model was taken as our researching material. Western blot, histological and electrophysiological techniques were used as the studying tools. The results showed that the expression level of GSK-3ß and CRMP-2 were elevated after seizure induction, and the administration of lovastatin reversed this effect and significantly reduced the extent of MFS in both DG and CA3 region in the hippocampus. The alteration of expression level of GSK-3ß and CRMP-2 after seizure induction proposes that GSK-3ß and CRMP-2 are crucial for MFS and epiletogenesis. The fact that lovastatin reversed the expression level of GSK-3ß and CRMP-2 indicated that GSK-3ß and CRMP-2 are possible to be a novel mechanism of lovatstain to suppress MFS and revealed a new therapeutic target and researching direction for studying the mechanism of MFS and epileptogenesis.
Asunto(s)
Giro Dentado/efectos de los fármacos , Modelos Animales de Enfermedad , Glucógeno Sintasa Quinasa 3/metabolismo , Lovastatina/uso terapéutico , Fibras Musgosas del Hipocampo/efectos de los fármacos , Pilocarpina/toxicidad , Estado Epiléptico/prevención & control , Animales , Anticolesterolemiantes/uso terapéutico , Western Blotting , Giro Dentado/enzimología , Giro Dentado/patología , Electrofisiología , Epilepsia del Lóbulo Temporal/inducido químicamente , Epilepsia del Lóbulo Temporal/prevención & control , Glucógeno Sintasa Quinasa 3 beta , Péptidos y Proteínas de Señalización Intercelular , Masculino , Fibras Musgosas del Hipocampo/enzimología , Fibras Musgosas del Hipocampo/patología , Agonistas Muscarínicos/toxicidad , Proteínas del Tejido Nervioso/metabolismo , Ratas , Ratas Wistar , Convulsiones/inducido químicamente , Convulsiones/prevención & control , Estado Epiléptico/inducido químicamenteRESUMEN
Carisbamate (CRS, RWJ-333369) is a novel antiepileptic drug awaiting approval for use in the treatment of partial and generalized seizures. Our aim was to determine whether CRS modulates synaptic transmission in the dentate gyrus (DG) and the underlying mechanism. The whole-cell patch-clamp method was used to record AMPA receptor- and NMDA receptor-mediated excitatory postsynaptic currents (EPSC(AMPA) and EPSC(NMDA)) and GABA(A) receptor-mediated inhibitory postsynaptic currents (IPSCs) in granule cells of the DG in brain slices prepared from 3- to 5-week-old male Wistar rats. CRS (30-300 µM) inhibited the evoked EPSC(AMPA) and EPSC(NMDA) by the same extent (20%) with significantly altered CV(-2), suggesting presynaptic modulation. It did not significantly change the inward currents induced by AMPA application. The inhibitory effect of CRS on the evoked EPSC(AMPA) was not occluded by selective voltage-gated Ca(2+) channel blockers, ruling out the involvement of presynaptic Ca(2+) channels. The frequency, but not the amplitude, of spontaneous EPSC(AMPA) was significantly reduced by CRS. However, CRS did not alter either the frequency or the amplitude of TTX-insensitive miniature EPSC(AMPA), indicating an action potential-dependent mechanism was involved. In addition, CRS (100 or 300 µM) did not significantly change the amplitude of the evoked IPSCs. To summarize, our results suggest that CRS reduces glutamatergic transmission by an action potential-dependent presynaptic mechanism and consequently inhibits excitatory synaptic strength in the DG without affecting GABAergic transmission. This effect may contribute to the antiepileptic action observed clinically at therapeutic concentrations of CRS.
Asunto(s)
Anticonvulsivantes/farmacología , Carbamatos/farmacología , Giro Dentado/citología , Ácido Glutámico/farmacología , Neuronas/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , 2-Amino-5-fosfonovalerato/farmacología , 6-Ciano 7-nitroquinoxalina 2,3-diona/farmacología , Animales , Biofisica , Bloqueadores de los Canales de Calcio/farmacología , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Antagonistas del GABA/farmacología , Técnicas In Vitro , Masculino , Técnicas de Placa-Clamp , Picrotoxina/farmacología , Ratas , Ratas Wistar , omega-Conotoxina GVIA/farmacologíaRESUMEN
BACKGROUND AND PURPOSE: Levetiracetam is an effective anti-epileptic drug in the treatment of partial and generalized seizure. The purpose of the present study was to investigate whether levetiracetam regulates AMPA and NMDA receptor-mediated excitatory synaptic transmission and to determine its site of action in the dentate gyrus (DG), the area of the hippocampus that regulates seizure activities. EXPERIMENTAL APPROACH: Whole-cell patch-clamp method was used to record the AMPA and NMDA receptor-mediated excitatory postsynaptic currents (EPSC(AMPA) and EPSC(NMDA)) in the presence of specific antagonists, from the granule cells in the DG in brain slice preparations from young Wistar rats (60-120 g). KEY RESULTS: Levetiracetam (100 microM) inhibited both evoked EPSC(AMPA) and EPSC(NMDA) to an equal extent (80%), altered the paired-pulse ratio (from 1.39 to 1.25), decreased the frequency of asynchronous EPSC and prolonged the inter-event interval of miniature EPSC(AMPA) (from 2.7 to 4.6 s) without changing the amplitude, suggesting a presynaptic action of levetiracetam. The inhibitory effect of levetiracetam on evoked EPSC(AMPA) was blocked by omega-agatoxin TK (100 nM), a selective P/Q-type voltage-dependent calcium channel blocker, but not by nimodipine (10 microM) or omega-conotoxin (400 nM). CONCLUSIONS AND IMPLICATIONS: These results suggest that levetiracetam modulated the presynaptic P/Q-type voltage-dependent calcium channel to reduce glutamate release and inhibited the amplitude of EPSC in DG. This effect is most likely to contribute to the anti-epileptic action of levetiracetam in patients.
Asunto(s)
Anticonvulsivantes/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Ácido Glutámico/efectos de los fármacos , Piracetam/análogos & derivados , Animales , Canales de Calcio Tipo P/efectos de los fármacos , Canales de Calcio Tipo P/metabolismo , Canales de Calcio Tipo Q/efectos de los fármacos , Canales de Calcio Tipo Q/metabolismo , Giro Dentado/efectos de los fármacos , Giro Dentado/metabolismo , Ácido Glutámico/metabolismo , Técnicas In Vitro , Levetiracetam , Masculino , Técnicas de Placa-Clamp , Piracetam/farmacología , Ratas , Ratas Wistar , Receptores AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismoRESUMEN
PURPOSE: The dentate gyrus (DG) is a gateway that regulates seizure activity in the hippocampus. We investigated the site of action of lamotrigine (LTG), an effective anticonvulsant, in the regulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and N-methyl-D-aspartic acid (NMDA) receptor-mediated excitatory synaptic transmission on DG. METHODS: Evoked AMPA and NMDA receptor-mediated excitatory postsynaptic currents (eEPSCampa and eEPSCnmda) were recorded by whole-cell patch-clamp recording from the granule cells of DG in brain slice preparation of young Wistar rats (60-120 g). Exogenously applied AMPA and NMDA-induced currents and AMPA receptor-mediated miniature EPSC (mEPSCampa) were recorded in the presence of specific antagonists. RESULTS: LTG inhibited both eEPSCampa and eEPSCnmda, and had no effect on exogenously applied NMDA-induced current indicating LTG inhibited glutamate release. Previous studies demonstrated that alteration in glutamate concentration in synaptic cleft causes parallel changes of eEPSCampa and eEPSCnmda. Our results showed that LTG inhibited eEPSCampa significantly more than eEPSCnmda (p < 0.05), suggesting that LTG may also have blocked the postsynaptic AMPA receptor. The hypothesis is further supported by the facts that; (1) LTG (30-100 microM) inhibited direct exogenously applied AMPA-induced currents (to 90%), (2) LTG significantly reduced the amplitude, but not the frequency of mEPSCampa and asynchronous (EPSC), and (3) LTG-induced reduction of eEPSCampa was not associated with a modification of the paired-pulse ratio. To sum up, LTG exerts a postsynaptic inhibitory mechanism on the AMPA receptor. CONCLUSIONS: Our results demonstrate that LTG suppresses postsynaptic AMPA receptors and reduces glutamate release in granule cells of DG. The postsynaptic effect can be one of the underlying mechanisms of LTG's anticonvulsant action.
Asunto(s)
Anticonvulsivantes/farmacología , Giro Dentado/metabolismo , Epilepsia/tratamiento farmacológico , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Glutamatos/metabolismo , Receptores AMPA/efectos de los fármacos , Triazinas/farmacología , Animales , Anticonvulsivantes/uso terapéutico , Giro Dentado/efectos de los fármacos , Modelos Animales de Enfermedad , Epilepsia/fisiopatología , Potenciales Postsinápticos Excitadores/fisiología , Glutamatos/efectos de los fármacos , Lamotrigina , Masculino , Ratas , Ratas Wistar , Receptores AMPA/agonistas , Receptores AMPA/metabolismo , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/metabolismo , Triazinas/uso terapéutico , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/farmacologíaRESUMEN
Levetiracetam (LEV) is an effective antiepileptic drug (AED) with distinct mechanism from the conventional AEDs. The major physiological function of ROMK1 channels is to maintain the resting membrane potential (RMP). In this study, we investigated the mechanisms underling LEV on ROMK1 channels. Xenopus oocytes were injected with mRNA to express the wild-type or mutant ROMK1 channels. Giant inside-out patch clamp recordings were performed to study the effect of LEV on these channels. LEV increased the activity of ROMK1 channels in a concentration-dependent manner and enhanced both wild-type and pH(i) gating residue mutant (K80M) channels over a range of pH(i) values. LEV activated the mutated channels at PIP(2)-binding sites (R188Q, R217A and K218A) and PKC-phosphorylation sites channels (S4A, S183A, T191A, T193A, S201A and T234A). However, this drug failed to enhance the channel activity in the presence of PKA inhibitors and did not activate the mutants of PKA-phosphorylation sites on C-terminal (S219A, S313A) and the constructed mutants (S219D and S313D) that mimic the negative charge carried by a phosphate group bound to a serine. Our results demonstrated PKA-mediated phosphorylation is a novel mechanism for LEV activating ROMK1 channels. These findings show that LEV activates ROMK1 channels independently from pH(i) and not via a PIP(2)- or PKC-dependent pathway. The effects of LEV may come from the PKA-induced conformational change but not charge-charge interaction in ROMK1 channels. Enhancing the activity of ROMK1 channels may be an important molecular mechanism for the antiepileptic effects of LEV in restoring neuronal RMP to prevent seizure spreading.
Asunto(s)
Anticonvulsivantes/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Piracetam/análogos & derivados , Canales de Potasio de Rectificación Interna/fisiología , Animales , Células Cultivadas , Concentración de Iones de Hidrógeno , Levetiracetam , Mutagénesis Sitio-Dirigida , Oocitos/efectos de los fármacos , Oocitos/fisiología , Técnicas de Placa-Clamp , Fosfatidilinositol 4,5-Difosfato/fisiología , Fosforilación , Piracetam/farmacología , ARN Mensajero/genética , XenopusRESUMEN
Parkinson's disease (PD) is a neurodegenerative disease that mainly affects dopaminergic (DA-ergic) neurons in the substantia nigra pars compacta (SNc). Glutamate modulates neuronal excitability, and a high concentration of glutamatergic receptors is found on DA-ergic neurons in the SNc. Paraquat (PQ) is a putative causative agent for PD. Its effects on synaptic glutamate transmission in SNc DA-ergic neurons were evaluated using whole-cell voltage-clamp recording in brain slices from 7- to 14-day-old Wistar rats. In the presence of bicuculline (BIC), strychnine, and dl-aminophosphonovaleric acid, PQ reversibly suppressed AMPA receptor-mediated evoked excitatory postsynaptic currents (eEPSCs) in a concentration-dependent manner (P<0.05). In the presence of tetrodotoxin (1 microM), PQ (50 microM) significantly reduced the amplitudes, but not the frequencies, of miniature EPSCs in the SNc, suggesting PQ inhibited eEPSCs through a postsynaptic mechanism. Exogenous application of AMPA to induce AMPA-mediated inward currents excluded involvement of a presynaptic response. The AMPA-induced currents in the SNc were significantly reduced by PQ (50 microM) to 74% of control levels (P<0.05), supporting that PQ acts on postsynaptic AMPA receptors. No effect of PQ on eEPSCs was seen in the LD thalamic nucleus and hippocampus, showing PQ specifically inhibited DA-ergic neurons in the SNc. Our results demonstrate a novel mechanism of action of PQ on glutamate-gated postsynaptic AMPA receptors in SNc DA-ergic neurons. This effect may attenuate the excitability and function of DA-ergic neurons in the SNc, which may contribute to the pathogenesis of PD.
Asunto(s)
Dopamina , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Neuronas/efectos de los fármacos , Paraquat/farmacología , Receptores AMPA/antagonistas & inhibidores , Sustancia Negra/efectos de los fármacos , Animales , Animales Recién Nacidos , Dopamina/fisiología , Potenciales Postsinápticos Excitadores/fisiología , Femenino , Masculino , Neuronas/metabolismo , Neuronas/fisiología , Ratas , Ratas Wistar , Receptores AMPA/fisiología , Sustancia Negra/metabolismoRESUMEN
Mossy fiber sprouting (MFS) is the main characteristic of temporal lobe epilepsy (TLE), which is highly correlated with the frequencies of recurrent seizures as well as degrees of severity of TLE. A recent MRI technique, referred to as diffusion spectrum imaging (DSI), can resolve crossing fibers and investigate the intravoxel heterogeneity of water molecular diffusion. Being able to achieve higher accuracy in depicting the complex fiber architecture, DSI may help improve localization of the seizure-induced epileptic foci. In this study, two indices of DSI, which represented the mean diffusivity (MSL) and diffusion anisotropy (DA), were proposed. A correlative study between diffusion characteristics and the severity of MFS was investigated in the pilocarpine-induced status epilepticus (SE) rat model. Nine SE rats and five control rats were studied with MRI and histological Timm's staining. For MSL, no significant correlation was found in the dentate gyrus (DG), r=-0.36; p=0.2017, and positive correlation was found in cornu ammonis (CA3), r=0.62; p=0.0174. The correlation between DA and Timm's score showed positive correlation in DG, r=0.71; p=0.0047, and negative correlation in CA3, r=-0.63; p=0.0151. Our results were compatible with the previous reports on fiber architecture alterations in DG and CA3 subregions. In conclusion, the histological correspondence of DSI indices was demonstrated. With DSI indices, longitudinal follow-up of hippocampal fiber architecture can be achieved to elucidate the pathophysiology of TLE, which might be helpful in disease localization.