RESUMEN
GABA(A) receptors are critically involved in hippocampal oscillations. GABA(A) receptor α1 and α2 subunits are differentially expressed throughout the hippocampal circuitry and thereby may have distinct contributions to oscillations. It is unknown which GABA(A) receptor α subunit controls hippocampal oscillations and where these receptors are expressed. To address these questions we used transgenic mice expressing GABA(A) receptor α1 and/or α2 subunits with point mutations (H101R) that render these receptors insensitive to allosteric modulation at the benzodiazepine binding site, and tested how increased or decreased function of α subunits affects hippocampal oscillations. Positive allosteric modulation by zolpidem prolonged decay kinetics of hippocampal GABAergic synaptic transmission and reduced the frequency of cholinergically induced oscillations. Allosteric modulation of GABAergic receptors in CA3 altered oscillation frequency in CA1, while modulation of GABA receptors in CA1 did not affect oscillations. In mice having a point mutation (H101R) at the GABA(A) receptor α2 subunit, zolpidem effects on cholinergically induced oscillations were strongly reduced compared to wild-type animals, while zolpidem modulation was still present in mice with the H101R mutation at the α1 subunit. Furthermore, genetic knockout of α2 subunits strongly reduced oscillations, whereas knockout of α1 subunits had no effect. Allosteric modulation of GABAergic receptors was strongly reduced in unitary connections between fast spiking interneurons and pyramidal neurons in CA3 of α2H101R mice, but not of α1H101R mice, suggesting that fast spiking interneuron to pyramidal neuron synapses in CA3 contain α2 subunits. These findings suggest that α2-containing GABA(A) receptors expressed in the CA3 region provide the inhibition that controls hippocampal rhythm during cholinergically induced oscillations.
Asunto(s)
Región CA3 Hipocampal/fisiología , Receptores de GABA-A/fisiología , Animales , Cognición/fisiología , Agonistas de Receptores de GABA-A/farmacología , Técnicas In Vitro , Interneuronas/fisiología , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Células Piramidales/fisiología , Piridinas/farmacología , ZolpidemRESUMEN
The hippocampus is critical for a wide range of emotional and cognitive behaviors. Here, we performed the first genome-wide search for genes influencing hippocampal oscillations. We measured local field potentials (LFPs) using 64-channel multi-electrode arrays in acute hippocampal slices of 29 BXD recombinant inbred mouse strains. Spontaneous activity and carbachol-induced fast network oscillations were analyzed with spectral and cross-correlation methods and the resulting traits were used for mapping quantitative trait loci (QTLs), i.e., regions on the genome that may influence hippocampal function. Using genome-wide hippocampal gene expression data, we narrowed the QTLs to eight candidate genes, including Plcb1, a phospholipase that is known to influence hippocampal oscillations. We also identified two genes coding for calcium channels, Cacna1b and Cacna1e, which mediate presynaptic transmitter release and have not been shown to regulate hippocampal network activity previously. Furthermore, we showed that the amplitude of the hippocampal oscillations is genetically correlated with hippocampal volume and several measures of novel environment exploration.
Asunto(s)
Estudios de Asociación Genética , Hipocampo/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/genética , Animales , Carbacol/farmacología , Análisis por Conglomerados , Electrodos , Regulación de la Expresión Génica/efectos de los fármacos , Hipocampo/efectos de los fármacos , Técnicas In Vitro , Patrón de Herencia/efectos de los fármacos , Patrón de Herencia/genética , Locomoción/efectos de los fármacos , Locomoción/genética , Ratones , Ratones Endogámicos , Red Nerviosa/efectos de los fármacos , Red Nerviosa/fisiología , Tamaño de los Órganos/efectos de los fármacos , Tamaño de los Órganos/genética , Sitios de Carácter Cuantitativo/efectos de los fármacos , Sitios de Carácter Cuantitativo/genética , Carácter Cuantitativo HeredableRESUMEN
Megalencephalic leucoencephalopathy with subcortical cysts is a genetic brain disorder with onset in early childhood. Affected infants develop macrocephaly within the first year of life, after several years followed by slowly progressive, incapacitating cerebellar ataxia and spasticity. From early on, magnetic resonance imaging shows diffuse signal abnormality and swelling of the cerebral white matter, with evidence of highly increased white matter water content. In most patients, the disease is caused by mutations in the gene MLC1, which encodes a plasma membrane protein almost exclusively expressed in brain and at lower levels in leucocytes. Within the brain, MLC1 is mainly located in astrocyte-astrocyte junctions adjacent to the blood-brain and cereborspinal fluid-brain barriers. Thus far, the function of MLC1 has remained unknown. We tested the hypothesis that MLC1 mutations cause a defect in ion currents involved in water and ion homeostasis, resulting in cerebral white matter oedema. Using whole-cell patch clamp studies we demonstrated an association between MLC1 expression and anion channel activity in different cell types, most importantly astrocytes. The currents were absent in chloride-free medium and in cells with disease-causing MLC1 mutations. MLC1-dependent currents were greatly enhanced by hypotonic pretreatment causing cell swelling, while ion channel blockers, including Tamoxifen, abolished the currents. Down regulation of endogenous MLC1 expression in astrocytes by small interfering RNA greatly reduced the activity of this channel, which was rescued by overexpression of normal MLC1. The current-voltage relationship and the pharmacological profiles of the currents indicated that the channel activated by MLC1 expression is a volume-regulated anion channel. Such channels are involved in regulatory volume decrease. We showed that regulatory volume decrease was hampered in lymphoblasts from patients with megalencephalic leucoencephalopathy. A similar trend was observed in astrocytes with decreased MLC1 expression; this effect was rescued by overexpression of normal MLC1. In the present study, we show that absence or mutations of the MLC1 protein negatively impact both volume-regulated anion channel activity and regulatory volume decrease, indicating that megalencephalic leucoencephalopathy is caused by a disturbance of cell volume regulation mediated by chloride transport.
Asunto(s)
Astrocitos/patología , Cloruros/metabolismo , Quistes/fisiopatología , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/fisiopatología , Transporte Iónico/fisiología , Proteínas de la Membrana/genética , Astrocitos/metabolismo , Tamaño de la Célula , Quistes/metabolismo , Quistes/patología , Células HEK293 , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/metabolismo , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/patología , Humanos , Proteínas de la Membrana/metabolismoRESUMEN
Ongoing neuronal oscillations in vivo exhibit non-random amplitude fluctuations as reflected in a slow decay of temporal auto-correlations that persist for tens of seconds. Interestingly, the decay of auto-correlations is altered in several brain-related disorders, including epilepsy, depression and Alzheimer's disease, suggesting that the temporal structure of oscillations depends on intact neuronal networks in the brain. Whether structured amplitude modulation occurs only in the intact brain or whether isolated neuronal networks can also give rise to amplitude modulation with a slow decay is not known. Here, we examined the temporal structure of cholinergic fast network oscillations in acute hippocampal slices. For the first time, we show that a slow decay of temporal correlations can emerge from synchronized activity in isolated hippocampal networks from mice, and is maximal at intermediate concentrations of the cholinergic agonist carbachol. Using zolpidem, a positive allosteric modulator of GABA(A) receptor function, we found that increased inhibition leads to longer oscillation bursts and more persistent temporal correlations. In addition, we asked if these findings were unique for mouse hippocampus, and we therefore analysed cholinergic fast network oscillations in rat prefrontal cortex slices. We observed significant temporal correlations, which were similar in strength to those found in mouse hippocampus and human cortex. Taken together, our data indicate that fast network oscillations with temporal correlations can be induced in isolated networks in vitro in different species and brain areas, and therefore may serve as model systems to investigate how altered temporal correlations in disease may be rescued with pharmacology.
Asunto(s)
Carbacol/farmacología , Agonistas Colinérgicos/farmacología , Hipocampo/anatomía & histología , Hipocampo/fisiología , Potenciales de la Membrana/fisiología , Red Nerviosa/efectos de los fármacos , Red Nerviosa/fisiología , Animales , Relación Dosis-Respuesta a Droga , Electrofisiología , Agonistas de Receptores de GABA-A/farmacología , Hipocampo/efectos de los fármacos , Humanos , Masculino , Ratones , Ratones Endogámicos DBA , Red Nerviosa/anatomía & histología , Periodicidad , Corteza Prefrontal/anatomía & histología , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/fisiología , Piridinas/farmacología , Ratas , Ratas Wistar , ZolpidemRESUMEN
Cognitive ability and the properties of brain oscillation are highly heritable in humans. Genetic variation underlying oscillatory activity might give rise to differences in cognition and behavior. How genetic diversity translates into altered properties of oscillations and synchronization of neuronal activity is unknown. To address this issue, we investigated cellular and synaptic mechanisms of hippocampal fast network oscillations in eight genetically distinct inbred mouse strains. The frequency of carbachol-induced oscillations differed substantially between mouse strains. Since GABAergic inhibition sets oscillation frequency, we studied the properties of inhibitory synaptic inputs (IPSCs) received by CA3 and CA1 pyramidal cells of three mouse strains that showed the highest, lowest and intermediate frequencies of oscillations. In CA3 pyramidal cells, the frequency of rhythmic IPSC input showed the same strain differences as the frequency of field oscillations. Furthermore, IPSC decay times in both CA1 and CA3 pyramidal cells were faster in mouse strains with higher oscillation frequencies than in mouse strains with lower oscillation frequency, suggesting that differences in GABA(A)-receptor subunit composition exist between these strains. Indeed, gene expression of GABA(A)-receptor ß2 (Gabrb2) and ß3 (Gabrb2) subunits was higher in mouse strains with faster decay kinetics compared with mouse strains with slower decay kinetics. Hippocampal pyramidal neurons in mouse strains with higher oscillation frequencies and faster decay kinetics fired action potential at higher frequencies. These data indicate that differences in genetic background may result in different GABA(A)-receptor subunit expression, which affects the rhythm of pyramidal neuron firing and fast network activity through GABA synapse kinetics.
RESUMEN
Endocannabinoids control hippocampal inhibitory synaptic transmission through activation of presynaptic CB(1) receptors. During depolarization-induced suppression of inhibition (DSI), endocannabinoids are synthesized upon postsynaptic depolarization. The endocannabinoid 2-arachidonoylglycerol (2-AG) may mediate hippocampal DSI. Currently, the best studied pathway for biosynthesis of 2-AG involves the enzyme diacylglycerol lipase (DAGL). However, whether DAGL is necessary for hippocampal DSI is controversial and was not systematically addressed. Here, we investigate DSI at unitary connections between CB(1) receptor-containing interneurons and pyramidal neurons in CA1. We found that the novel DAGL inhibitor OMDM-188, as well as the established inhibitor RHC-80267, did not affect DSI. As reported previously, effects of the DAGL inhibitor tetrahydrolipstatin depended on the application method: postsynaptic intracellular application left DSI intact, while incubation blocked DSI. We show that all DAGL inhibitors tested block slow self-inhibition in neocortical interneurons, which involves DAGL. We conclude that DAGL is not involved in DSI at unitary connections in hippocampus.
Asunto(s)
Potenciales Postsinápticos Inhibidores/fisiología , Lipoproteína Lipasa/metabolismo , Inhibición Neural/fisiología , Neuronas/fisiología , Animales , Animales Recién Nacidos , Benzoxazinas/farmacología , Ciclohexanonas/farmacología , Estimulación Eléctrica/métodos , Inhibidores Enzimáticos/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Antagonistas del GABA/farmacología , Proteínas Fluorescentes Verdes/genética , Hipocampo/citología , Técnicas In Vitro , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Lipoproteína Lipasa/antagonistas & inhibidores , Lisina/análogos & derivados , Lisina/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Morfolinas/farmacología , Naftalenos/farmacología , Neocórtex/citología , Inhibición Neural/efectos de los fármacos , Inhibición Neural/genética , Neuronas/efectos de los fármacos , Piridazinas/farmacología , Quinoxalinas/farmacología , Receptor Cannabinoide CB1/agonistas , Receptor Cannabinoide CB1/deficiencia , Valina/análogos & derivados , Valina/farmacologíaRESUMEN
Primary sensory cortical areas continuously receive thalamic inputs that arrive at different frequencies depending on the amount of sensory activity. The cortical response to repeated sensory stimuli rapidly adapts and different frequencies recruit cortical neuronal networks to different extents. GABAergic inhibition limits the spread of excitation within cortical neuronal networks. However, it is unknown how frequency adaptation of cortical network activity at different frequencies is shaped by GABAergic inhibition. Here, we find that in acute slices of visual cortex area V1 GABAergic inhibition affects frequency adaptation depending on the frequency of activity. Using voltage-sensitive dye imaging, we found that while increasing inhibitory postsynaptic currents (IPSCs) with flunitrazepam dampened the spread of cortical excitation, short-term adaptations to different stimulation frequencies were differentially affected. At high frequencies (40 Hz), facilitation of cortical excitation was no longer transient, but facilitation was sustained. At low frequencies (10 Hz) flunitrazepam decreased a depression of the excitation. In contrast, in mice lacking the GABA(A) receptor alpha1 subunit facilitation was reduced and depression enhanced. These findings suggest that GABAergic inhibition affects cortical excitation at different frequencies differentially, favoring facilitation at higher frequencies of excitation.
Asunto(s)
Adaptación Fisiológica/fisiología , Potenciales Postsinápticos Inhibidores/fisiología , Plasticidad Neuronal/fisiología , Corteza Visual/fisiología , Ácido gamma-Aminobutírico/metabolismo , Animales , Estimulación Eléctrica , Ratones , Ratones Endogámicos C57BL , Imagen de Colorante Sensible al VoltajeRESUMEN
A major challenge in neuroscience is to identify genes that influence specific behaviors and to understand the intermediary neuronal mechanisms. One approach is to identify so-called endophenotypes at different levels of neuronal organization from synapse to brain activity. An endophenotype is a quantitative trait that is closer to the gene action than behavior, and potentially a marker of neuronal mechanisms underlying behavior. Hippocampal activity and, in particular, hippocampal oscillations have been suggested to underlie various cognitive and motor functions. To identify quantitative traits that are potentially useful for identifying genes influencing hippocampal activity, we measured gamma oscillations and spontaneous activity in acute hippocampal slices from eight inbred mouse strains under three experimental conditions. We estimated the heritability of more than 200 quantitative traits derived from this activity. We observed significant differences between the different mouse strains, particularly in the amplitude of the activity and the correlation between activities in different hippocampal subregions. Interestingly, these traits had a low genetic correlation between the three experimental conditions, which suggests that different genetic components influence the activity in different conditions. Our findings show that several traits of hippocampal gamma oscillations and spontaneous activity are heritable and could thus be potentially useful in gene-finding strategies based on endophenotypes.
Asunto(s)
Hipocampo/fisiología , Ratones Endogámicos/genética , Ratones Endogámicos/fisiología , Red Nerviosa/fisiología , Neuronas/fisiología , Fenotipo , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/genética , Potenciales de Acción/fisiología , Análisis de Varianza , Animales , Carbacol/farmacología , Análisis por Conglomerados , Electrofisiología , Análisis de Fourier , Agonistas del GABA/farmacología , Hipocampo/efectos de los fármacos , Ratones , Microelectrodos , Red Nerviosa/efectos de los fármacos , Neuronas/efectos de los fármacos , Oscilometría , Periodicidad , Piridinas/farmacología , Receptores de GABA-A/fisiología , Especificidad de la Especie , ZolpidemRESUMEN
Human brain oscillations occur in different frequency bands that have been linked to different behaviours and cognitive processes. Even within specific frequency bands such as the beta- (14-30 Hz) or gamma-band (30-100 Hz), oscillations fluctuate in frequency and amplitude. Such frequency fluctuations most probably reflect changing states of neuronal network activity, as brain oscillations arise from the correlated synchronized activity of large numbers of neurons. However, the neuronal mechanisms governing the dynamic nature of amplitude and frequency fluctuations within frequency bands remain elusive. Here we show that in acute slices of rat prefrontal cortex (PFC), carbachol-induced oscillations in the beta-band show frequency and amplitude fluctuations. Fast and slow non-harmonic frequencies are distributed differentially over superficial and deep cortical layers, with fast frequencies being present in layer 3, while layer 6 only showed slow oscillation frequencies. Layer 5 pyramidal cells and interneurons experience both fast and slow frequencies and they time their spiking with respect to the dominant frequency. Frequency and phase information is encoded and relayed in the layer 5 network through timed excitatory and inhibitory synaptic transmission. Our data indicate that frequency fluctuations in the beta-band reflect synchronized activity in different cortical subnetworks, that both influence spike timing of output layer 5 neurons. Thus, amplitude and frequency fluctuations within frequency bands may reflect activity in distinct cortical neuronal subnetworks that may process information in a parallel fashion.
Asunto(s)
Potenciales de Acción/fisiología , Relojes Biológicos/fisiología , Red Nerviosa/fisiología , Plasticidad Neuronal/fisiología , Neuronas/fisiología , Corteza Prefrontal/fisiología , Animales , Animales Recién Nacidos , Células Cultivadas , Ratas , Ratas WistarRESUMEN
Encoding and retention of information in memory are associated with a sustained increase in the amplitude of neuronal oscillations for up to several seconds. We reasoned that coordination of oscillatory activity over time might be important for memory and, therefore, that the amplitude modulation of oscillations may be abnormal in Alzheimer disease (AD). To test this hypothesis, we measured magnetoencephalography (MEG) during eyes-closed rest in 19 patients diagnosed with early-stage AD and 16 age-matched control subjects and characterized the autocorrelation structure of ongoing oscillations using detrended fluctuation analysis and an analysis of the life- and waiting-time statistics of oscillation bursts. We found that Alzheimer's patients had a strongly reduced incidence of alpha-band oscillation bursts with long life- or waiting-times (< 1 s) over temporo-parietal regions and markedly weaker autocorrelations on long time scales (1-25 seconds). Interestingly, the life- and waiting-times of theta oscillations over medial prefrontal regions were greatly increased. Whereas both temporo-parietal alpha and medial prefrontal theta oscillations are associated with retrieval and retention of information, metabolic and structural deficits in early-stage AD are observed primarily in temporo-parietal areas, suggesting that the enhanced oscillations in medial prefrontal cortex reflect a compensatory mechanism. Together, our results suggest that amplitude modulation of neuronal oscillations is important for cognition and that indices of amplitude dynamics of oscillations may prove useful as neuroimaging biomarkers of early-stage AD.
Asunto(s)
Enfermedad de Alzheimer/fisiopatología , Lóbulo Parietal/fisiología , Corteza Prefrontal/fisiopatología , Anciano , Estudios de Casos y Controles , HumanosRESUMEN
The amplitude fluctuations of ongoing oscillations in the electroencephalographic (EEG) signal of the human brain show autocorrelations that decay slowly and remain significant at time scales up to tens of seconds. We call these long-range temporal correlations (LRTC). Abnormal LRTC have been observed in several brain pathologies, but it has remained unknown whether genetic factors influence the temporal correlation structure of ongoing oscillations. We recorded the ongoing EEG during eyes-closed rest in 390 monozygotic and dizygotic twins and investigated the temporal structure of ongoing oscillations in the alpha- and beta-frequency bands using detrended fluctuation analysis (DFA). The strength of LRTC was more highly correlated in monozygotic than in dizygotic twins. Statistical analysis attributed up to approximately 60% of the variance in DFA to genetic factors, indicating a high heritability for the temporal structure of amplitude fluctuations in EEG oscillations. Importantly, the DFA and EEG power were uncorrelated. LRTC in ongoing oscillations are robust, heritable, and independent of power, suggesting that LRTC and oscillation power are governed by distinct biophysical mechanisms and serve different functions in the brain. We propose that the DFA method is an important complement to classical spectral analysis in fundamental and clinical research on ongoing oscillations.
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Relojes Biológicos/genética , Relojes Biológicos/fisiología , Encéfalo/fisiología , Electroencefalografía/métodos , Gemelos Dicigóticos/fisiología , Gemelos Monocigóticos/fisiología , Adolescente , Adulto , Femenino , Humanos , Masculino , Países Bajos , Sistema de Registros , Procesamiento de Señales Asistido por Computador , Factores de Tiempo , Gemelos Dicigóticos/genética , Gemelos Monocigóticos/genéticaRESUMEN
Diacylglycerol (DAG) is a prominent endogenous modulator of synaptic transmission. Recent studies proposed two apparently incompatible pathways, via protein kinase C (PKC) and via Munc13. Here we show how these two pathways converge. First, we confirm that DAG analogs indeed continue to potentiate transmission after PKC inhibition (the Munc13 pathway), but only in neurons that previously experienced DAG analogs, before PKC inhibition started. Second, we identify an essential PKC pathway by expressing a PKC-insensitive Munc18-1 mutant in munc18-1 null mutant neurons. This mutant supported basic transmission, but not DAG-induced potentiation and vesicle redistribution. Moreover, synaptic depression was increased, but not Ca2+-independent release evoked by hypertonic solutions. These data show that activation of both PKC-dependent and -independent pathways (via Munc13) are required for DAG-induced potentiation. Munc18-1 is an essential downstream target in the PKC pathway. This pathway is of general importance for presynaptic plasticity.
Asunto(s)
Diglicéridos/fisiología , Plasticidad Neuronal/fisiología , Proteína Quinasa C/fisiología , Receptores Presinapticos/fisiología , Transducción de Señal/fisiología , Animales , Corteza Cerebral/citología , Corteza Cerebral/efectos de los fármacos , Células Cromafines/metabolismo , Diglicéridos/metabolismo , Electrofisiología , Inhibidores Enzimáticos/farmacología , Femenino , Hipocampo/citología , Hipocampo/efectos de los fármacos , Cinética , Lentivirus/genética , Ratones , Ratones Noqueados , Microscopía Electrónica , Proteínas Munc18/genética , Proteínas Munc18/metabolismo , Mutación/fisiología , Neuronas/citología , Neuronas/metabolismo , Neuronas/ultraestructura , Técnicas de Placa-Clamp , Ésteres del Forbol/farmacología , Fosforilación , Embarazo , Proteína Quinasa C/antagonistas & inhibidores , Receptores Presinapticos/ultraestructuraRESUMEN
Nicotine enhances attention and working memory by activating nicotinic acetylcholine receptors (nAChRs). The prefrontal cortex (PFC) is critical for these cognitive functions and is also rich in nAChR expression. Specific cellular and synaptic mechanisms underlying nicotine's effects on cognition remain elusive. Here we show that nicotine exposure increases the threshold for synaptic spike-timing-dependent potentiation (STDP) in layer V pyramidal neurons of the mouse PFC. During coincident presynaptic and postsynaptic activity, nicotine reduces dendritic calcium signals associated with action potential propagation by enhancing GABAergic transmission. This results from a series of presynaptic actions involving different PFC interneurons and multiple nAChR subtypes. Pharmacological block of nAChRs or GABA(A) receptors prevented nicotine's actions and restored STDP, as did increasing dendritic calcium signals with stronger postsynaptic activity. Thus, by activating nAChRs distributed throughout the PFC neuronal network, nicotine affects PFC information processing and storage by increasing the amount of postsynaptic activity necessary to induce STDP.
Asunto(s)
Potenciales de Acción/efectos de los fármacos , Plasticidad Neuronal/fisiología , Neuronas/fisiología , Nicotina/farmacología , Agonistas Nicotínicos/farmacología , Corteza Prefrontal/citología , Animales , Animales Recién Nacidos , Señalización del Calcio/efectos de los fármacos , Señalización del Calcio/fisiología , Interacciones Farmacológicas , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Antagonistas del GABA/farmacología , Técnicas In Vitro , Ratones , Ratones Endogámicos C57BL , Modelos Neurológicos , Red Nerviosa/efectos de los fármacos , Red Nerviosa/fisiología , Plasticidad Neuronal/efectos de los fármacos , Neuronas/clasificación , Tiempo de Reacción/efectos de los fármacos , Tiempo de Reacción/fisiología , Receptores Nicotínicos/genética , Receptores Nicotínicos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
Prompt recovery after intense activity is an essential feature of most mammalian synapses. Here we show that synapses with reduced expression of the presynaptic gene munc18-1 suffer from increased depression during intense stimulation at glutamatergic, GABAergic, and neuromuscular synapses. Conversely, munc18-1 overexpression makes these synapses recover faster. Concomitant changes in the readily releasable vesicle pool and its refill kinetics were found. The number of vesicles docked at the active zone and the total number of vesicles per terminal correlated with both munc18-1 expression levels and the size of the releasable vesicle pool. These data show that varying expression of a single gene controls synaptic recovery by modulating the number of docked, release-ready vesicles and thereby replenishment of the secretion capacity.
Asunto(s)
Proteínas Munc18/metabolismo , Vesículas Sinápticas/metabolismo , Animales , Regulación de la Expresión Génica , Heterocigoto , Ratones , Ratones Transgénicos , Microscopía Electrónica , Proteínas Munc18/genética , Transmisión Sináptica , Vesículas Sinápticas/genética , Vesículas Sinápticas/ultraestructura , Factores de TiempoRESUMEN
We investigated to what extent Pitx3 deficiency, causing hyperdopaminergic transmission in the nucleus accumbens microcircuitry, may lead to developmental changes. First, spontaneous firing activity of cholinergic interneurons in the nucleus accumbens was recorded in vitro. Firing patterns in the Pitx3-deficient mice were more variable and intrinsically different from those observed in wild-type mice. Next, to test whether the irregular firing patterns observed in mutant mice affected the endogenous nicotinic modulation of the GABAergic input of medium spiny neurons, we recorded spontaneous GABAergic inputs to these cells before and after the application of the nicotinic receptor blocker mecamylamine. Effects of mecamylamine were found in slices of either genotype, but in a rather inconsistent manner. Possibly this was attributable to heterogeneity in firing of nearby cholinergic interneurons. Thus paired recordings of cholinergic interneurons and medium spiny neurons were performed to more precisely control the experimental conditions of the cholinergic modulation of GABAergic synaptic transmission. We found that controlling action potential firing in cholinergic neurons leads to a conditional increase in GABAergic input frequency in wild-type mice but not in Pitx3-deficient mice. We conclude that Pitx3-deficient mice have neural adaptations at the level of the nucleus accumbens microcircuitry that in turn may have behavioral consequences. It is discussed to what extent dopamine release in the nucleus accumbens may be a long-term gating mechanism leading to alterations in cholinergic transmission in the nucleus accumbens, in line with previously reported neural adaptations found as consequences of repeated drug treatment in rodents.
Asunto(s)
Fibras Colinérgicas/fisiología , Proteínas de Homeodominio/genética , Plasticidad Neuronal , Núcleo Accumbens/crecimiento & desarrollo , Núcleo Accumbens/fisiología , Sinapsis/fisiología , Factores de Transcripción/genética , Ácido gamma-Aminobutírico/fisiología , Potenciales de Acción/fisiología , Animales , Dopamina/metabolismo , Regulación de la Expresión Génica , Proteínas de Homeodominio/fisiología , Interneuronas/fisiología , Mecamilamina/farmacología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Antagonistas Nicotínicos/farmacología , Técnicas de Placa-Clamp , Receptores Nicotínicos/efectos de los fármacos , Receptores Nicotínicos/fisiología , Sinapsis/química , Transmisión Sináptica/fisiología , Factores de Transcripción/fisiología , Ácido gamma-Aminobutírico/análisisRESUMEN
We here investigated inhibitory synapse turnover in the adult brain using the hypothalamic supraoptic nucleus where new synapses form during different physiological conditions, in particular on oxytocin neurons largely controlled by GABAergic inputs and locally released oxytocin. Patch clamp recordings and ultrastructural analysis of the nucleus in acute slices from late gestating rats showed that oxytocin and estrogen promoted rapid formation of inhibitory synapses. Thus, after 2-h exposure to a combination of oxytocin and 17-beta estradiol, the frequency of miniature inhibitory postsynaptic currents was significantly enhanced. Since their amplitude and presynaptic GABA release probability were unmodified, this indicated an increased number of synapses. Electron microscopy confirmed increased densities of symmetric, putative GABAergic synapses within 2-h exposure to the peptide or steroid, effects which were reversible and oxytocin receptor mediated. Our observations thus offer direct evidence that hypothalamic GABAergic microcircuitries can undergo rapid and functional remodeling under changing neuroendocrine conditions.
Asunto(s)
Estrógenos/farmacología , Neuronas/efectos de los fármacos , Oxitocina/farmacología , Núcleo Supraóptico/efectos de los fármacos , Sinapsis/metabolismo , Ácido gamma-Aminobutírico/metabolismo , Animales , Estrógenos/metabolismo , Femenino , Técnicas In Vitro , Neuronas/citología , Neuronas/metabolismo , Oxitocina/metabolismo , Técnicas de Placa-Clamp , Embarazo , Ratas , Ratas Wistar , Núcleo Supraóptico/citología , Núcleo Supraóptico/metabolismo , Sinapsis/química , Sinapsis/efectos de los fármacos , Sinapsis/ultraestructuraRESUMEN
RATIONALE: Nicotine affects many aspects of human cognition, including attention and memory. Activation of nicotinic acetylcholine receptors (nAChRs) in neuronal networks modulates activity and information processing during cognitive tasks, which can be observed in electroencephalograms (EEGs) and functional magnetic resonance imaging studies. OBJECTIVES: In this review, we will address aspects of nAChR functioning as well as synaptic and cellular modulation important for nicotinic impact on neuronal networks that ultimately underlie its effects on cognition. Although we will focus on general mechanisms, an emphasis will be put on attention behavior and nicotinic modulation of prefrontal cortex. In addition, we will discuss how nicotinic effects at the neuronal level could be related to its effects on the cognitive level through the study of electrical oscillations as observed in EEGs and brain slices. RESULTS/CONCLUSIONS: Very little is known about mechanisms of how nAChR activation leads to a modification of electrical oscillation frequencies in EEGs. The results of studies using pharmacological interventions and transgenic animals implicate some nAChR types in aspects of cognition, but neuronal mechanisms are only poorly understood. We are only beginning to understand how nAChR distribution in neuronal networks impacts network functioning. Unveiling receptor and neuronal mechanisms important for nicotinic modulation of cognition will be instrumental for treatments of human disorders in which cholinergic signaling have been implicated, such as schizophrenia, attention deficit/hyperactivity disorder, and addiction.
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Encéfalo/efectos de los fármacos , Cognición/efectos de los fármacos , Red Nerviosa/efectos de los fármacos , Nicotina/farmacología , Receptores Nicotínicos/efectos de los fármacos , Animales , Animales Modificados Genéticamente , Atención/efectos de los fármacos , Electroencefalografía/efectos de los fármacos , Humanos , Imagen por Resonancia Magnética , Memoria/efectos de los fármacos , Neuronas/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacosRESUMEN
There is a large variation in structurally and functionally different GABA(A) receptor subtypes. The expression pattern of GABA(A) receptor subunits is highly regulated, both temporarily and spatially. Especially during development, profound changes in subunit expression have been described. In most brain areas, the GABA(A) receptor alpha1 subunit replaces the alpha2 and/or alpha3 subunit as major alpha subunit. This is accompanied by a marked decrease in the open time of GABA(A) receptors and hence in the duration of postsynaptic responses. We describe here the development of GABAergic, synaptic transmission in mice lacking the alpha1 subunit. We show that alpha1 is to a large extent--but not entirely--responsible for the relatively short duration of postsynaptic responses in the developing and the mature brain. However, alpha1 already affects GABAergic transmission in the neonatal cerebral cortex when it is only sparsely expressed. It appears that the alpha1 -/- mice do not show a large reduction in GABAergic synapses but do retain long-lasting postsynaptic currents into adulthood. Hence, they form a good model to study the functional role of developmental GABA(A) receptor subunit switching.
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Inhibición Neural/fisiología , Neuronas/fisiología , Receptores de GABA-A/deficiencia , Sinapsis/fisiología , Transmisión Sináptica/fisiología , Factores de Edad , Anestésicos Locales/farmacología , Animales , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Estimulación Eléctrica/métodos , Potenciales de la Membrana/fisiología , Potenciales de la Membrana/efectos de la radiación , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas/efectos de la radiación , Técnicas de Placa-Clamp/métodos , ARN Mensajero/biosíntesis , Receptores de GABA-A/genética , Receptores de GABA-A/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Tetrodotoxina/farmacología , Corteza Visual/citología , Corteza Visual/crecimiento & desarrollo , Corteza Visual/efectos de la radiaciónRESUMEN
Repeated exposure to drugs of abuse causes persistent behavioral sensitization and associated adaptations of striatal neurotransmission, which is thought to play an important role in certain aspects of drug addiction. Microdialysis and neurochemical studies suggest that intermittent morphine treatment may lead to a long-term increase in both ACh and dopaminergic neurotransmission in the nucleus accumbens (NAc). This implies that both cholinergic modulation of GABA synapses and their sensitivity to dopaminergic transmission might be changed, ultimately leading to a modified NAc output. Here we investigate to what extent cholinergic modulation and sensitivity to amphetamine, causing endogenous dopamine efflux, of GABAergic transmission in the nucleus accumbens are affected 3 weeks after a period of daily morphine injections in adult rats. To this end, we recorded medium spiny neurons using whole cell voltage clamp and monitored the frequency and amplitude of spontaneous GABAergic synaptic currents. We observed that the effect of nicotine on the frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) was suppressed in rats pretreated with morphine, whereas the effects of mecamylamine and tetrodotoxin (TTX) were increased. These results indicate that the probability of GABA release was increased and that this effect resulted from an upregulation of the endogenous activation of presynaptic nicotinic receptors. In addition, we observed an increased sensitivity to in vitro application of amphetamine. This suggests that the long-term increase in dopaminergic transmission caused by the morphine treatment affects GABA synapses in the NAc. Hence, there may be two parallel synaptic mechanisms by which drugs of abuse may affect processing and integration of NAc inputs.
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Acetilcolina/metabolismo , Morfina/farmacología , Núcleo Accumbens/citología , Sinapsis/efectos de los fármacos , Ácido gamma-Aminobutírico/metabolismo , Anfetamina/farmacología , Anestésicos Locales/farmacología , Animales , Estimulantes del Sistema Nervioso Central/farmacología , Interacciones Farmacológicas , Técnicas In Vitro , Masculino , Mecamilamina/farmacología , Potenciales de la Membrana/efectos de los fármacos , Narcóticos/farmacología , Inhibición Neural/efectos de los fármacos , Nicotina/farmacología , Agonistas Nicotínicos/farmacología , Antagonistas Nicotínicos/farmacología , Núcleo Accumbens/efectos de los fármacos , Ratas , Ratas Wistar , Sinapsis/metabolismo , Tetrodotoxina/farmacologíaRESUMEN
Expression, functional properties, and clustering of alpha 1-, alpha 2-, and alpha 3-subunit containing GABA(A) receptors (GABA(A)Rs) were studied in dorsomedial SON neurons of the adult female rat supraoptic nucleus (SON) around parturition. We show that, although the decay time constant (tau(decay)) of GABAergic postsynaptic currents between and within individual recordings was very diverse, ranging from fast (i.e., alpha 1-like) to significantly slower (i.e., non-alpha 1-like), there was an overall shift towards slower decaying synaptic currents during the onset of lactation. This shift is not due to changes in mRNA expression levels, because real-time quantitative PCR assays indicated that the relative contribution of alpha 1, alpha 2, and alpha 3 remained the same before and after parturition. Also, changes in phosphorylation levels are not likely to affect the tau(decay) of postsynaptic currents. In alpha-latrotoxin (alpha-LTX)-induced bursts of synaptic currents from individual synapses, the tau(decay) of consecutive synaptic events within bursts was very similar, but between bursts there were large differences in tau(decay). This suggested that different synapses within individual SON neurons contain distinct GABA(A)R subtypes. Using multilabeling confocal microscopy, we examined the distribution of postsynaptic alpha 1-, alpha 2-, and alpha 3-GABA(A)Rs, based on colocalization with gephyrin. We show that the three GABA(A)R subtypes occurred either in segregated clusters of one subtype as well as in mixed clusters of two or possibly even three receptor subtypes. After parturition, the density and proportion of clusters containing alpha 2- (or alpha 3-), but not alpha1-GABA(A)Rs, was significantly increased. Thus, the functional synaptic diversity at the postsynaptic level in dorsomedial SON neurons is correlated with a differential clustering of distinct GABA(A)R subtypes at individual synapses.