RESUMO
Amyloid-ß oligomers (Aßo) are the most pathologically relevant Aß species in Alzheimer's disease (AD), because they induce early synaptic dysfunction that leads to learning and memory impairments. In contrast, increasing VEGF (Vascular Endothelial Growth Factor) brain levels have been shown to improve learning and memory processes, and to alleviate Aß-mediated synapse dysfunction. Here, we designed a new peptide, the blocking peptide (BP), which is derived from an Aßo-targeted domain of the VEGF protein, and investigated its effect on Aß-associated toxicity. Using a combination of biochemical, 3D and ultrastructural imaging, and electrophysiological approaches, we demonstrated that BP strongly interacts with Aßo and blocks Aß fibrillar aggregation process, leading to the formation of Aß amorphous aggregates. BP further impedes the formation of structured Aßo and prevents their pathogenic binding to synapses. Importantly, acute BP treatment successfully rescues long-term potentiation (LTP) in the APP/PS1 mouse model of AD, at an age when LTP is highly impaired in hippocampal slices. Moreover, BP is also able to block the interaction between Aßo and VEGF, which suggests a dual mechanism aimed at both trapping Aßo and releasing VEGF to alleviate Aßo-induced synaptic damage. Our findings provide evidence for a neutralizing effect of the BP on Aß aggregation process and pathogenic action, highlighting a potential new therapeutic strategy.
Assuntos
Doença de Alzheimer , Peptídeos beta-Amiloides , Camundongos , Animais , Peptídeos beta-Amiloides/toxicidade , Peptídeos beta-Amiloides/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Doença de Alzheimer/metabolismo , Memória/fisiologia , Aprendizagem , Sinapses/metabolismo , Fragmentos de Peptídeos/farmacologia , Fragmentos de Peptídeos/metabolismoRESUMO
Vascular endothelial growth factor (VEGF) is known to be required for the action of antidepressant therapies but its impact on brain synaptic function is poorly characterized. Using a combination of electrophysiological, single-molecule imaging and conditional transgenic approaches, we identified the molecular basis of the VEGF effect on synaptic transmission and plasticity. VEGF increases the postsynaptic responses mediated by the N-methyl-D-aspartate type of glutamate receptors (GluNRs) in hippocampal neurons. This is concurrent with the formation of new synapses and with the synaptic recruitment of GluNR expressing the GluN2B subunit (GluNR-2B). VEGF induces a rapid redistribution of GluNR-2B at synaptic sites by increasing the surface dynamics of these receptors within the membrane. Consistently, silencing the expression of the VEGF receptor 2 (VEGFR2) in neural cells impairs hippocampal-dependent synaptic plasticity and consolidation of emotional memory. These findings demonstrated the direct implication of VEGF signaling in neurons via VEGFR2 in proper synaptic function. They highlight the potential of VEGF as a key regulator of GluNR synaptic function and suggest a role for VEGF in new therapeutic approaches targeting GluNR in depression.
Assuntos
Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Comportamento Animal , Técnicas de Cultura de Células , Potenciais Pós-Sinápticos Excitadores , Medo , Hipocampo/metabolismo , Camundongos , Plasticidade Neuronal/fisiologia , Neurônios/metabolismo , Subunidades Proteicas , Receptores de Glutamato/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapses/metabolismo , Transmissão Sináptica/fisiologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/fisiologiaRESUMO
In psychiatric and neurological diseases, sleep is often perturbed. Moreover, recent works on humans and animals tend to show that sleep plays a strong role in memory processes. Reciprocally, sleep dynamics following a learning task is modified [Hubert, Nature (London) 02663, 1 (2004), Peigneux, Neuron 44, 535 (2004)]. However, sleep analysis in humans and animals is often limited to the total sleep and wake duration quantification. These two parameters are not fully able to characterize the sleep dynamics. In mammals sleep presents a complex organization with an alternation of slow wave sleep (SWS) and paradoxical sleep (PS) episodes. Moreover, it has been shown recently that these sleep episodes are frequently interrupted by micro-arousal (without awakening). We present here a detailed analysis of the basal sleep properties emerging from the mechanisms underlying the vigilance states alternation in an animal model. These properties present a self-organized critical system signature and reveal the existence of two W, two SWS, and a PS structure exhibiting a criticality as met in sand piles. We propose a theoretical model of the sleep dynamics based on several interacting neuronal populations. This new model of sleep dynamics presents the same properties as experimentally observed, and explains the variability of the collected data. This experimental and theoretical study suggests that sleep dynamics shares several common features with critical systems.
RESUMO
Microtubules are key cytoskeletal components in the cytoplasm of eukaryotic cells where they have pleiotropic and vital roles in functions such as cell division, trafficking or morphogenesis. Microtubules are especially abundant in neurons. Although microtubules are in many cells dynamic polymers, they exhibit an extreme state of stability in neurons. Previous work has indicated a central role of microtubule associated proteins called STOPs in neuronal microtubule stabilization. We have recently developed STOP null mice. These mice were devoid of stable brain microtubules but to our surprise had nevertheless an apparently normal brain anatomy. However the mice showed synaptic defects affecting different forms of long- and short-term synaptic plasticity. These synaptic defects were associated with severe behavioral defects that showed a remarkable sensitivity to long-term treatment with neuroleptics. We discuss the relationship of the phenotypes observed in STOP null mice with current models of schizophrenia in which the multiple, severe, and neuroleptic sensitive mental disorders caused by the disease are due to a "disease of the synapse".
Assuntos
Citoesqueleto/fisiologia , Transtornos Mentais/patologia , Microtúbulos/fisiologia , Neurônios/ultraestrutura , Animais , Antipsicóticos/farmacologia , Antipsicóticos/uso terapêutico , Modelos Animais de Doenças , Humanos , Transtornos Mentais/tratamento farmacológico , Camundongos , Camundongos Knockout , Proteínas Associadas aos Microtúbulos/deficiência , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/fisiologia , Modelos Biológicos , Modelos Neurológicos , Plasticidade Neuronal , Esquizofrenia/tratamento farmacológico , Esquizofrenia/patologia , Tubulina (Proteína)/metabolismoRESUMO
It has been shown recently that in mitral cells of the rat olfactory bulb, N-methyl-D-aspartate (NMDA) autoreceptors are activated during mitral cell firing. Here we consider in more details the mechanisms of mitral cell self-excitation and its physiological relevance. We show that both ionotropic NMDA and non-NMDA autoreceptors are activated by glutamate released from primary and secondary dendrites. In contrast to non-NMDA autoreceptors, NMDA autoreceptors are almost exclusively located on secondary dendrites and their activation generates a large and sustained self-excitation. Both intracellularly evoked and miniature NMDA-R mediated synaptic potentials are blocked by intracellular bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) and result from a calcium-dependent release of glutamate. Self-excitation can be produced by a single spike, and trains of spikes result in frequency facilitation. Thus activation of excitatory autoreceptors is a major function of action potentials backpropagating in mitral cell dendrites, which results in an immediate positive feedback counteracting recurrent inhibition and increasing the signal-to-noise ratio of olfactory inputs.
Assuntos
Autorreceptores/metabolismo , Dendritos/metabolismo , Ácido Glutâmico/metabolismo , Bulbo Olfatório/metabolismo , Transdução de Sinais/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Sinalização do Cálcio/fisiologia , Estimulação Elétrica , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Antagonistas de Receptores de GABA-A , Técnicas In Vitro , Inibição Neural/efeitos dos fármacos , Inibição Neural/fisiologia , Bulbo Olfatório/citologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Receptores de AMPA/antagonistas & inibidores , Receptores de AMPA/metabolismo , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Receptores de Glutamato Metabotrópico/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/metabolismo , Sódio/metabolismoRESUMO
Several lines of evidence have suggested that decreases in postsynaptic inhibition may have a role in epileptogenesis in cortical structures. However, other studies have suggested that GABAergic inhibition is spared, or even augmented in some forms of post-lesional epilepsy. In the studies described here, inhibitory events were recorded in two models of post-lesional chronic epileptogenesis. (i) As previously reported (D.A. Prince and G.-F. Tseng. J. Neurophysiol. 69: 1276-1291. 1993), epileptiform activity develops in slices from partially isolated rat neocortical islands 2-3 weeks after the initial in vivo lesion. In this model of post-traumatic epilepsy, large amplitude polyphasic inhibitory postsynaptic currents (IPSCs) in layer V pyramidal neurons are associated with each interictal epileptiform field potential. The frequency of spontaneous IPSCs as well as miniature IPSCs was significantly increased in neocortical slices from the epileptogenic chronically injured cortex versus controls. Immunocytochemical reactions for parvalbumin and calbindin, calcium binding proteins present in subgroups of GABAergic neurons, showed an increased staining of both neuropil and somata within the epileptogenic tissue. Immunoreactivity for glutamic acid decarboxylase (GAD) and GABA also appeared to be increased in the neuropil. (ii) Cortical microgyri resembling human malformations were produced by freeze lesions made transcranially in P0 rat cortex (K.M. Jacobs, M.J. Gutnick, and D.A. Prince. Cereb. Cortex, 6: 514-523. 1996). The boundary between the four-layered microgyrus and surrounding cortex become epileptogenic within about 2 weeks, as judged by evoked extracellular field potentials and cellular activities. Epileptogenesis in the surrounding cortex is not altered when the microgyrus itself is isolated by transcortical cuts. Patch-clamp recordings from layer V neurons in the epileptogenic zone showed that spontaneous IPSCs are larger and more dependent on glutamatergic synapses than in control neurons. The amplitudes of polysynaptic IPSCs evoked by threshold stimulation were also larger than in control cells. Although evaluation of inhibitory events in these models is still incomplete, results to date suggest that GABAergic inhibition may be enhanced in epileptogenic areas associated with chronic cortical injury. Sprouting of axonal arborizations of pyramidal cells onto interneurons, upregulation of GABAergic neurons, and perhaps sprouting of inhibitory axons that make increased numbers of contacts onto pyramidal cells may all contribute to the increased inhibitory drive. Results in these models do not support the disinhibitory hypothesis of chronic epileptogenesis.
Assuntos
Córtex Cerebral/fisiologia , Epilepsias Parciais/etiologia , Epilepsia Pós-Traumática/etiologia , Inibição Neural , Ácido gama-Aminobutírico/fisiologia , Animais , Humanos , Células Piramidais/fisiologia , RatosRESUMO
The classical view of fast chemical synaptic transmission is that released neurotransmitter acts locally on postsynaptic receptors and is cleared from the synaptic cleft within a few milliseconds by diffusion and by specific reuptake mechanisms. This rapid clearance restricts the spread of neurotransmitter and, combined with the low affinities of many ionotropic receptors, ensures that synaptic transmission occurs in a point-to-point fashion. We now show, however, that when transmitter release is enhanced at hippocampal mossy fibre synapses, the concentration of glutamate increases and its clearance is delayed; this allows it to spread away from the synapse and to activate presynaptic inhibitory metabotropic glutamate receptors (mGluRs). At normal levels of glutamate release during low-frequency activity, these presynaptic receptors are not activated. When glutamate concentration is increased by higher-frequency activity or by blocking glutamate uptake, however, these receptors become activated, leading to a rapid inhibition of transmitter release. This effect may be related to the long-term depression of mossy fibre synaptic responses that has recently been shown after prolonged activation of presynaptic mGluRs (refs 2, 3). The use-dependent activation of presynaptic mGluRs that we describe here thus represents a negative feedback mechanism for controlling the strength of synaptic transmission.
Assuntos
Glutamatos/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Sinapses/metabolismo , Animais , Potenciais Evocados , Antagonistas de Aminoácidos Excitatórios/farmacologia , Cobaias , Hipocampo/metabolismo , Técnicas In Vitro , Ácidos Pimélicos/farmacologia , Quinoxalinas/farmacologia , Receptores de AMPA/antagonistas & inibidores , Receptores de AMPA/metabolismo , Receptores de Glutamato Metabotrópico/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/metabolismoRESUMO
A single mossy fiber input contains several release sites and is located on the proximal portion of the apical dendrite of CA3 neurons. It is, therefore, well suited to exert a strong influence on pyramidal cell excitability. Accordingly, the mossy fiber synapse has been referred to as a detonator or teacher synapse in autoassociative network models of the hippocampus. The very low firing rates of granule cells [Jung, M. W. & McNaughton, B. L. (1993) Hippocampus 3, 165-182], which give rise to the mossy fibers, raise the question of how the mossy fiber synapse temporally integrates synaptic activity. We have therefore addressed the frequency dependence of mossy fiber transmission and compared it to associational/commissural synapses in the CA3 region of the hippocampus. Paired pulse facilitation had a similar time course, but was 2-fold greater for mossy fiber synapses. Frequency facilitation, during which repetitive stimulation causes a reversible growth in synaptic transmission, was markedly different at the two synapses. At associational/ commissural synapses facilitation occurred only at frequencies greater than once every 10 s and reached a magnitude of about 125% of control. At mossy fiber synapses, facilitation occurred at frequencies as low as once every 40 s and reached a magnitude of 6-fold. Frequency facilitation was dependent on a rise in intraterminal Ca2+ and activation of Ca2+/calmodulin-dependent kinase II, and was greatly reduced at synapses expressing mossy fiber long-term potentiation. These results indicate that the mossy fiber synapse is able to integrate granule cell spiking activity over a broad range of frequencies, and this dynamic range is substantially reduced by long-term potentiation.
Assuntos
Hipocampo/fisiologia , Plasticidade Neuronal , Neurônios/fisiologia , Células Piramidais/fisiologia , Receptores de AMPA/fisiologia , Sinapses/fisiologia , 4-Aminopiridina/farmacologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Animais , Cálcio/farmacologia , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Ácido Egtázico/análogos & derivados , Ácido Egtázico/farmacologia , Potenciais Evocados/efeitos dos fármacos , Cobaias , Técnicas In Vitro , Potenciação de Longa Duração , Magnésio/farmacologia , Fibras Nervosas/efeitos dos fármacos , Fibras Nervosas/fisiologia , Quinoxalinas/farmacologia , Receptores de AMPA/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/fisiologia , Fatores de TempoRESUMO
Extracellular field potential recordings from the CA3 region in guinea pig hippocampal slices were used to study the release and action of dynorphin at the mossy fiber synapse. Dynorphin A(1-17) or U69593 inhibited mossy fiber synaptic responses in preparations in which the CA3 region was surgically isolated from the rest of the hippocampus. This inhibition was completely reversed by the kappa 1 selective antagonist nor-BNI, thus establishing the presence of functional kappa 1 receptors in CA3. Inhibitory effects of dynorphin on mossy fiber responses were unaltered in the presence of the N- or P-type Ca2+ channel blockers, omega-CgTx or omega-Aga IVA, respectively. This indicates that the action of dynorphin is independent of the particular type of Ca2+ channel that mediates transmitter release at the mossy fiber terminal. Heterosynaptic inhibition of mossy fiber responses was observed in the presence of nifedipine, omega-CgTx, or omega-Aga IVA, indicating that dynorphin release does not depend specifically on L-, N-, or P-type Ca2+ channels. The blockade of heterosynaptic inhibition by the membrane-permeant Ca2+ chelator EGTA-AM suggests the involvement of a slow Ca(2+)-dependent process in dynorphin release. On the basis of a variety of experimental evidence, we propose that the time course of heterosynaptic inhibition is determined primarily by the time course of clearance of dynorphin in the extracellular space.
Assuntos
Dinorfinas/farmacologia , Hipocampo/efeitos dos fármacos , Fibras Nervosas/efeitos dos fármacos , Sinapses/efeitos dos fármacos , Animais , Soluções Tampão , Cálcio/metabolismo , Canais de Cálcio/fisiologia , Dinorfinas/metabolismo , Cobaias , Hipocampo/metabolismo , Técnicas In Vitro , Membranas Intracelulares/metabolismo , Modelos Neurológicos , Inibição Neural , Receptores Opioides kappa/metabolismo , Fatores de TempoRESUMO
The N-methyl-D-aspartate receptor-independent form of long-term potentiation (LTP) at hippocampal mossy fiber synapses requires presynaptic Ca(2+)-dependent activation of adenylyl cyclase. To determine whether this form of LTP might occur at other synapses, we examined cerebellar parallel fibers that, like hippocampal mossy fiber synapses, express high levels of the Ca2+/calmodulin-sensitive adenylyl cyclase I. Repetitive stimulation of parallel fibers caused a long-lasting increase in synaptic strength that was associated with a decrease in paired-pulse facilitation. Blockade of glutamate receptors did not prevent LTP induction, nor did loading of Purkinje cells with a Ca2+ chelator. LTP was occluded by forskolin-induced potentiation and blocked by the protein kinase A inhibitor Rp-8-CPT-cAMPS. These findings suggest that parallel fiber synapses express a form of LTP that is dependent on the activation of a presynaptic adenylyl cyclase and is indistinguishable from LTP at hippocampal mossy fiber synapses.
Assuntos
Cerebelo/fisiologia , AMP Cíclico/fisiologia , Potenciação de Longa Duração/fisiologia , Fibras Nervosas/fisiologia , Sinapses/fisiologia , Adenilil Ciclases/metabolismo , Animais , Cálcio/farmacologia , Calmodulina/farmacologia , Colforsina/farmacologia , AMP Cíclico/análogos & derivados , AMP Cíclico/farmacologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Técnicas In Vitro , Ratos , Tionucleotídeos/farmacologiaRESUMO
1. Spontaneous inhibitory synaptic currents (sIPSCs) were studied with whole cell voltage-clamp recordings from 131 pyramidal cells in adult rat somatosensory cortical slices. Neurons were intracellulary labeled with biocytin and classified as supragranular (SG, layers 2-3), layer IV (IV), or infragranular (IG, layer V) on the basis of the laminar localization of their somata. Somatic areas were similar for SG, IV, and IG neurons. All identified pyramidal cells generated high-frequency gamma-aminobutyric acid (GABAA) receptor-mediated synaptic events. 2. Bath application of bicuculline blocked the sIPSCs and resulted in a decrease of approximately 0.5 nS in resting conductance and an inward shift in baseline current. 3. sIPSC frequency was significantly lower in SG versus IG or IV neurons, and this difference was accounted for by the occurrence of a higher percentage of bursts of sIPSCs in the IG and IV neurons. 4. Bath application of the alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) decreased the frequency of sIPSCs by 13-21%. By contrast, application of the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonovaleric acid (D-AP5) generally had no effect on spontaneous IPSC frequency, suggesting that AMPA rather than NMDA receptor activation contributed to resting discharge of inhibitory interneurons. 5. Addition of tetrodotoxin (TTX) to the perfusion medium reduced the spontaneous IPSC frequency by approximately 30-55%. The miniature IPSCs (mIPSCs) seen in TTX-containing solutions had a frequency of approximately 10 Hz and an average conductance of 0.42-0.48 nS. 6. The kinetic properties of mIPSCs generated in pyramidal cells of different layers were the same, with the rise times of approximately 0.9 ms and decay time constants of approximately 8 ms at a holding potential of 0 mV. The decay phase of mIPSCs was generally fitted by one exponential and displayed a voltage dependence with an e-fold increase in decay time constant for a every 198-mV depolarization. 7. These results show that there is ongoing spontaneous release of GABA in neocortical slices that gives rise to high-frequency impulse-related and non-impulse-related postsynaptic inhibitory currents. Activation of AMPA receptors on inhibitory interneurons accounts for only a small proportion of the GABAA receptor-mediated events. Judging from the distribution of mIPSC frequencies in neurons of different laminae, there is a relatively uniform distribution of inhibitory synapses throughout the cortex. Tonic activation of GABAA receptors on neocortical pyramidal neurons generates an increase in resting membrane conductance that may play an important role in vivo by preventing the development of hyperexcitability, modulating excitatory synaptic events, and controlling the rate and patterns of spike discharge.
Assuntos
Inibição Neural/fisiologia , Células Piramidais/fisiologia , Receptores de GABA-A/fisiologia , Córtex Somatossensorial/fisiologia , Transmissão Sináptica/fisiologia , Animais , Condutividade Elétrica , Potenciais Somatossensoriais Evocados/fisiologia , Técnicas In Vitro , Cinética , Potenciais da Membrana/fisiologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Córtex Somatossensorial/citologiaRESUMO
1. gamma-Aminobutyric acid-A (GABAA) receptor-mediated synaptic currents evoked by intracortical stimulation in rat somatosensory cortical slices maintained in vitro were studied using the whole cell patch-clamp technique. All anatomically identified pyramidal neurons of layer II-III (SG neurons), layer IV (IV neurons), and layer V (IG neurons) generated evoked inhibitory postsynaptic currents (eIPSCs) that were blocked by bicuculline. At threshold, eIPSCs had kinetic properties (rise time of 0.9 ms and decay time constant of 9 ms) similar to those of spontaneous IPSCs generated in the same cells. 2. The strength of inhibition was quantified by determining the stimulus threshold for evoking responses and the relationship between stimulus strength and eIPSC peak amplitudes (input/output curve). For eIPSCs recorded in control solution, the input/output curve was about four times steeper than for eIPSCs recorded in the presence of the ionotropic glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and D-2-amino-5-phosphonovalerate (D-AP5), suggesting the dependence of GABAA inhibition on synpatic excitation of interneurons. 3. In the presence of CNQX and D-AP5, monosynaptic IPSCs, evoked by stimulation close to the recording patch pipette, had similar input/output curves in SG and IG neurons. This suggests that the level of monosynaptic inhibition generated in these two populations of cells is similar. 4. When the stimulus was moved to a distant site > 350 microns from the recorded neuron, either in vertical or in horizontal direction, the stimulus intensity required for evoking IPSCs was higher, and the input/output curve was less steep. This suggests that the density of GABAergic somata and axons projecting to the recorded neuron is lower at these distances than at more proximal sites. 5. The maximum horizontal distance over which IPSCs could be evoked ("horizontal field") was larger in layer V than in other layers. The horizontal field (distance between stimulating and recording pipettes) was 600 microns in layer II-III, 580 microns in layer IV, and 720 microns in layer V. Anatomic identification of the somatosensory cortical barrels indicated that the extent of GABAergic projections was larger than the barrel hollow and might thus form a substrate for interbarrel inhibition in layer IV during cross-wisker stimulation. 6. The maximum vertical inhibitory field was larger than the maximum horizontal field. IPSCs could be evoked in layer V neurons by layer I stimuli, showing that a powerful interlaminar inhibition is present that may play a role in synchronizing the activity of neurons in a column. IPSCs evoked by layer I stimulation frequently had slower kinetics than those elicited by stimulation at sites close to the soma. 7. These findings suggest that functional GABAergic projections are characterized by a large degree of convergence. Quantification of GABAA-mediated IPSCs indicates that this zone of inhibitory synaptic convergence onto a given pyramidal neuron is subdivided into a powerful local inhibitory zone and a surrounding area of long-range, less effective, inhibitory projections. Potential roles for these concentric inhibitory areas in cortical processing of sensory information are discussed.
Assuntos
Mapeamento Encefálico/métodos , Potenciais Somatossensoriais Evocados/fisiologia , Inibição Neural/fisiologia , Células Piramidais/fisiologia , Receptores de GABA-A/fisiologia , Córtex Somatossensorial/fisiologia , Animais , Bicuculina/farmacologia , Potenciais Somatossensoriais Evocados/efeitos dos fármacos , Antagonistas GABAérgicos/farmacologia , Antagonistas de Receptores de GABA-A , Técnicas In Vitro , Técnicas de Patch-Clamp , Células Piramidais/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
We have used a number of approaches to address a possible role of metabotropic glutamate receptors (mGluRs) in mossy fiber long-term potentiation (LTP) in the hippocampus. We have used two types of mutant mice--one lacking the mGluR1 subtype of receptor and one lacking the gamma isoform of protein kinase C. In neither type of mouse did we find any alteration in the magnitude of mossy fiber LTP. We next examined whether mGluRs might modulate the magnitude and/or threshold for the induction of mossy fiber LTP. In these experiments we used tetani that were either just subthreshold or just suprathreshold for generating LTP. The mGluR antagonist (+)-alpha-methyl-4-carboxyphenylglycine [(+)MCPG] did not convert a subthreshold tetanus into a suprathreshold tetanus, nor did (+)MCPG have any effect on the small amount of LTP that was generated by a just suprathreshold tetanus. Based on our studies, we have been unable to identify a role for mGluRs in mossy fiber LTP.
Assuntos
Benzoatos/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Glicina/análogos & derivados , Hipocampo/fisiologia , Potenciação de Longa Duração/fisiologia , Receptores de Glutamato Metabotrópico/fisiologia , Animais , Glicina/farmacologia , Técnicas In Vitro , Camundongos , Camundongos Knockout , Proteína Quinase C/fisiologia , Receptores de Glutamato Metabotrópico/antagonistas & inibidoresRESUMO
Several behavioral studies in rat (Gallagher, 1988) have suggested that opioids in the hippocampus could play an important role in learning and memory. However, in this species, very few reports specifically address the issue of physiological actions of opioids released by the mossy fibers which constitute the principal source of dynorphin and enkephalin in the hippocampus. In the guinea pig high frequency stimulation of mossy fibers causes a transient heterosynaptic inhibition of neighboring mossy fibers (Weisskopf et al., 1993) or perforant path synapses in the dentate (Wagner et al., 1993), which is mediated by the synaptic release of dynorphin that activates presynaptic kappa receptors. We show here that neither exogenous nor endogenous dynorphin affect mossy fiber excitatory postsynaptic potentials in the Sprague-Dawley rat, which is consistent with the finding that kappa receptor binding in the mossy fiber termination zone is dense in the guinea pig and sparse in this rat. More surprisingly, although kappa receptor binding is found in the rat dentate gyrus molecular layer and in the CA3 pyramidal cell layer, dynorphin had no action on perforant path field responses, somatic potassium currents or evoked monosynaptic inhibitory postsynaptic currents in CA3 cells. This lack of action appears to be an exception among rodents as dynorphin significantly inhibited mossy fiber responses in the hamster, mouse, and even another strain of rat, Long-Evans. Unlike the kappa mediated actions, the mu opioid receptor agonist DAMGO inhibited Sprague-Dawley mossy fiber responses, as it does in guinea pig. In contrast to other investigators, however, we found that the opioid receptor antagonist naloxone had no effect on Sprague-Dawley mossy fiber LTP.
Assuntos
Dinorfinas/fisiologia , Cobaias/fisiologia , Hipocampo/fisiologia , Ratos/fisiologia , Animais , Cricetinae , Giro Denteado/metabolismo , Potenciação de Longa Duração , Masculino , Mesocricetus , Camundongos , Camundongos Endogâmicos C57BL , Fibras Nervosas/fisiologia , Vias Neurais/fisiologia , Células Piramidais/metabolismo , Ratos Sprague-Dawley , Receptores Opioides kappa/metabolismo , Transmissão SinápticaRESUMO
We examined whether the interconnections between areas 17 and 18 are spatially reciprocal, i.e., whether a column of cells in area 17 receives from the same region of area 18 as it sends projections to, and vice versa. We addressed this question by making side by side injections of retrograde fluorescent tracers in area 18, calculating the convergence and divergence of the connections from area 17 to 18. We compared these values with previously reported values of divergence and convergence of the projections from area 18 to area 17. The results demonstrate that there is a good match between the convergence and divergence of the area 17 to area 18 connection and, respectively, the divergence and convergence of the reverse connection. We confirmed directly the spatial reciprocity by injecting simultaneously in area 17 a retrograde and an anterograde tracer and by analyzing quantitatively the density of anterograde and retrograde labeling across the surface of area 18. There was an excellent match between the density maps of retrogradely labeled cells and anterogradely labeled axon terminals in area 18. Connections between areas 17 and 18 therefore exhibit large degrees of convergence and divergence and are spatially reciprocal. Thus, a given column of cells within one of these two areas is reciprocally interconnected with a large region of the opposite area. Such an organization may provide the basis for synchronization of firing of neurons across these two areas, as revealed by cross-correlation studies.
Assuntos
Córtex Visual/anatomia & histologia , Amidinas , Animais , Biotina/análogos & derivados , Mapeamento Encefálico , Gatos , Dextranos , Corantes Fluorescentes , RodaminasRESUMO
1. We used an in vitro model to explore critical aspects of chronic epileptogenesis. Partial neocortical isolations having intact blood supply were made in rat and guinea pig from postnatal day 7 to 34 and then examined 1 to 150 days later in standard brain slice preparations. 2. The epileptogenic potential of several different types of lesions was assessed. Slices containing transcortical (i.e., gray matter) lesions, with or without a contiguous white matter injury (i.e., "undercut"), developed chronic epileptogenesis after a latency of approximately 1-2 wk, manifested by evoked and spontaneous "interictal" discharges and evoked "ictal" events. The region of hyperexcitability did not extend beyond approximately 2 mm from the chronic transcortical lesion and was rarely observed in slices having only an apparent white matter injury. 3. Multiple recordings and current source density (CSD) analysis identified layer V as the source of the interictal discharge. 4. Significant differences in CSD profiles of the evoked interictal discharge occurred between chronically epileptogenic slices and control (noninjured) slices bathed in the convulsant, bicuculline methiodide, suggesting that mechanisms other than disinhibition must be involved in posttraumatic epileptogenesis. 5. Interictal events were blocked in most but not all chronically injured slices by application of the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonovalerate (D-AP5), suggesting that non-NMDA receptors were predominantly involved in some preparations. 6. This model of chronic epileptogenesis in vitro will be useful in studies relevant to mechanisms of posttraumatic epilepsy in man.
Assuntos
Senescência Celular/fisiologia , Córtex Cerebral/fisiopatologia , Epilepsia/fisiopatologia , Animais , Mapeamento Encefálico , Córtex Cerebral/lesões , Técnicas de Cultura , Epilepsia Pós-Traumática/fisiopatologia , Potenciais Evocados/fisiologia , Cobaias , Córtex Motor/lesões , Córtex Motor/fisiopatologia , Rede Nervosa/fisiopatologia , Regeneração Nervosa/fisiologia , Neurônios/fisiologia , Ratos , Ratos Sprague-Dawley , Tempo de Reação/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologia , Córtex Somatossensorial/lesões , Córtex Somatossensorial/fisiopatologia , Transmissão Sináptica/fisiologiaRESUMO
Visual cortical areas are richly but selectively connected by "patchy" projections. We characterized these connections physiologically with cross-correlograms (CCHs), calculated for neuron pairs or small groups located one each in visual areas 17 and 18 of the cat. The CCHs were then compared to the visuotopic and orientation match of the neurons' receptive fields (RFs). For both spontaneous and visually driven activity, most non-flat correlograms were centered; i.e. the most likely temporal relationship between spikes in the two areas is a synchronous one. Although spikes are most likely to occur simultaneously, area 17 spikes may occur before area 18 or vice versa, giving the cross-correlogram peak a finite width (temporal dispersion). Cross-correlograms fell into one of three groups according to their full-width at half peak height: 1-8 ms (modal width, 3 ms), 15-65 ms (modal width 30 ms), or 100-1000 ms (modal width 400 ms). These classificatory groups are nonoverlapping; the three types of coupling appeared singly and in combination. Neurons whose receptive fields (RFs) are nonoverlapping or cross-oriented may yet be coupled, but the coupling is more likely to be the broadest type of coupling than the medium-dispersed type. The sharpest type of coupling is found exclusively between neurons with at least partially overlapping RFs and mostly between neurons whose stimulus orientation preferences matched to within 22.5 deg. The maximum spatial dispersion observed in the RFs of coupled neurons compares well with the maximum divergence seen anatomically in the A18/A17 projection system. We suggest three different mechanisms to produce each of the three different degrees of observed spatial and temporal coherence. All mechanisms use common input of cortical origin. For medium and broad coupling, this common input arises from cell assemblies split between both sides of the 17/18 projection system, but acting synchronously. Such distributed common-input cell assemblies are a means of overcoming sparse connectivity and achieving synaptic transmission in the pyramidal network.