RESUMEN
The BK channel is a Ca(2+) and voltage-gated conductance responsible for shaping action potential waveforms in many types of neurons. Type II BK channels are differentiated from type I channels by their pharmacology and slow gating kinetics. The ß4 accessory subunit confers type II properties on BK α subunits. Empirically derived properties of BK channels, with and without the ß4 accessory subunit, were obtained using a heterologous expression system under physiological ionic conditions. These data were then used to study how BK channels alone (type I) and with the accessory ß4 subunit (type II) modulate action potential properties in biophysical neuron models. Overall, the models support the hypothesis that it is the slower kinetics provided by the ß4 subunit that endows the BK channel with type II properties, which leads to broadening of action potentials and, secondarily, to greater recruitment of SK channels reducing neuronal excitability. Two regions of parameter space distinguished type II and type I effects; one where the range of BK-activating Ca(2+) was high (>20 µM) and the other where BK-activating Ca(2+) was low (â¼0.4-1.2 µM). The latter required an elevated BK channel density, possibly beyond a likely physiological range. BK-mediated sharpening of the spike waveform associated with the lack of the ß4 subunit was sensitive to the properties of voltage-gated Ca(2+) channels due to electrogenic effects on spike duration. We also found that depending on Ca(2+) dynamics, type II BK channels may have the ability to contribute to the medium AHP, a property not generally ascribed to BK channels, influencing the frequency-current relationship. Finally, we show how the broadening of action potentials conferred by type II BK channels can also indirectly increase the recruitment of SK-type channels decreasing the excitability of the neuron.
Asunto(s)
Potenciales de Acción/fisiología , Canales de Potasio de Gran Conductancia Activados por el Calcio/fisiología , Modelos Neurológicos , Animales , Simulación por Computador , Células HEK293 , Humanos , Activación del Canal Iónico/fisiología , Ratones , Técnicas de Placa-ClampRESUMEN
Pyramidal neurons in the CA3 region of the hippocampal formation integrate synaptic information arriving in the dendrites within discrete laminar regions. At potentials near or below the resting potential integration of synaptic signals is most affected by the passive properties of the cell and hyperpolarization-activated currents (I(h)). Here we focused specifically on a subset of neurons within the CA3b subregion of the rat hippocampus in order to better understand their membrane response within subthreshold voltage ranges. Using a combined experimental and computational approach we found that the passive properties of these neurons varied up to fivefold between cells. Likewise, there was a large variance in the expression of I(h) channels. However, the contribution of I(h) was minimal at resting potentials endowing the membrane with an apparent linear response to somatic current injection within +/-10 mV. Unlike in CA1 pyramidal neurons, however, I(h) activation was not potentiated in an activity-dependent manner. Computer modeling, based on a combination of voltage- and current-clamp data, suggested that an increasing density of these channels with distance from the soma, compared with a uniform distribution, would have no significant effect on the general properties of the cell because of their relatively lower expression. Nonetheless, temporal summation of excitatory inputs was affected by the presence of I(h) in the dendrites in a frequency- and distance-dependent fashion.
Asunto(s)
Canales Catiónicos Regulados por Nucleótidos Cíclicos/fisiología , Hipocampo/fisiología , Potenciales de la Membrana/fisiología , Canales de Potasio/fisiología , Células Piramidales/fisiología , Transmisión Sináptica/fisiología , Animales , Hipocampo/citología , Canales Regulados por Nucleótidos Cíclicos Activados por Hiperpolarización , Técnicas In Vitro , Proteínas del Tejido Nervioso/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
In hippocampal pyramidal neurons from aged animals voltage-gated Ca2+ entry and the slow, post-burst afterhyperpolarization are enhanced. As a result, there is a decrease in neuronal excitability and, in turn, an alteration in synaptic plasticity. Restricting the caloric intake of a rodent is a well-known paradigm for increasing lifespan and ameliorating a number of neurodegenerative features of aging, including deficits in synaptic plasticity and cognition. Here we show in rat CA1 pyramidal neurons from aged animals (18-20 months old) that a restricted diet prevents the enhancement of dendritic spike-mediated Ca2+ accumulation. In contrast, no significant changes in the rates of Ca2+ recovery were observed suggesting that Ca2+ clearance mechanisms are not affected by aging or caloric restriction. Lastly, we found that caloric restriction also prevented the aging-associated increase in the slow, post-burst afterhyperpolarization. Our results suggest that caloric restriction-sensitive changes in Ca2+ accumulation and membrane excitability may in part account for the protective effects of dietary restriction on synaptic plasticity and learning deficits in aged animals.
Asunto(s)
Potenciales de Acción/fisiología , Envejecimiento/fisiología , Calcio/metabolismo , Restricción Calórica , Hipocampo/citología , Células Piramidales/fisiología , Potenciales de Acción/efectos de los fármacos , Análisis de Varianza , Animales , Dendritas/fisiología , Dendritas/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Estimulación Eléctrica/métodos , Técnicas In Vitro , Técnicas de Placa-Clamp/métodos , Células Piramidales/citología , Ratas , Ratas Endogámicas F344 , Factores de TiempoRESUMEN
The mechanisms underlying the inhibitory effects of dopamine (DA) on layer V pyramidal neuron excitability in the prelimbic region of the rat medial prefrontal cortex were investigated. Under control conditions, DA depressed both action potential generation (driven by somatic current injection) and input resistance (R(N)). The presence of GABA(A) receptor antagonists blocked DA-induced depression of action potential generation and revealed a delayed increase in excitability that persisted for the duration of experimental recording, up to 20 min following the washout of DA. In contrast to spike generation, disinhibition did not affect the transient depression of R(N) produced by DA, suggesting independent actions of DA on spike generation and R(N). Consistent with the hypothesis that DA acts to decrease pyramidal cell output via a GABAergic mechanism, DA increased the frequency of spontaneous inhibitory postsynaptic currents in both the absence and presence of TTX. Furthermore focal application of GABA to a perisomatic region mimicked the inhibitory effect of DA on spike production without affecting R(N). Focal application of bicuculline to the same location reversed the inhibitory effect of bath-applied DA on spike generation, while again having no effect on R(N). The depression of R(N) by DA was both occluded and mimicked by the Na(+) channel blocker TTX, suggesting the involvement of a Na(+) conductance in reducing pyramidal cell R(N) during the acute presence of DA. Together these data demonstrate that the acute presence of DA decreases pyramidal neuron excitability by two independent mechanisms. At the same time DA triggers a delayed and longer-lasting increase in excitability that is partially masked by synaptic inhibition.
Asunto(s)
Dopamina/farmacología , Corteza Prefrontal/citología , Corteza Prefrontal/fisiología , Células Piramidales/efectos de los fármacos , Células Piramidales/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Bicuculina/farmacología , Calcio/metabolismo , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Antagonistas del GABA/farmacología , Antagonistas de Receptores de GABA-A , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Técnicas de Cultivo de Órganos , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , Receptores de GABA-A/fisiología , Sodio/metabolismo , Tetrodotoxina/farmacología , Ácido gamma-Aminobutírico/farmacologíaRESUMEN
Protein synthesis inhibitors block the maintenance of NMDA receptor-dependent long-term potentiation (LTP) both in vivo and in vitro. Protein synthesis inhibitors block mossy fiber(MF) LTP maintenance in vitro, but little is known about the effect of protein synthesis inhibitors on either induction or maintenance in MF-LTP in vivo. Here we study the role of protein synthesis in the induction of long-term potentiation at the mossy fiber-CA3 hippocampal synapse in vivo in anesthetized rats. The protein synthesis inhibitor anisomycin was administered at different doses (0.04, 10, or 40 nmol) into area CA3 15 min before delivering high-frequency stimulation (two times at 100 Hz, 1 sec). Anisomycin blocked MF-LTP induction in a dose-dependent manner; both 40 and 10 nmol blocked MF-LTP induction, but a lower dose of 0.04 nmol was without effect. The inhibitory effect of anisomycin on protein synthesis was determined by measuring the incorporation of [(35)S]methionine into the newly synthesized proteins. Percentages of protein synthesis inhibition were determined by comparing [(35)S] incorporation of anisomycin-treated samples with vehicle controls. Doses of 0.04, 10, or 40 nmol of anisomycin produced 21, 82, or 83% inhibition of [(35)S]methionine incorporation, respectively. The effect of anisomycin was verified using a single dose of the protein synthesis inhibitor cycloheximide (40 nmol). Cycloheximide also blocked MF-LTP induction. These results suggest that protein synthesis plays an important role in the induction of mossy fiber long-term potentiation in vivo.
Asunto(s)
Anisomicina/administración & dosificación , Cicloheximida/administración & dosificación , Potenciación a Largo Plazo/efectos de los fármacos , Fibras Musgosas del Hipocampo/efectos de los fármacos , Inhibidores de la Síntesis de la Proteína/administración & dosificación , Potenciales de Acción/efectos de los fármacos , Animales , Calcio/metabolismo , Relación Dosis-Respuesta a Droga , Estimulación Eléctrica , Colorantes Fluorescentes , Fura-2 , Técnicas In Vitro , Líquido Intracelular/metabolismo , Masculino , Metionina/metabolismo , Microinyecciones , Fibras Musgosas del Hipocampo/fisiología , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-DawleyRESUMEN
We examined how biophysical properties and neuronal morphology affect the propagation of individual postsynaptic potentials (PSPs) from synaptic inputs to the soma. This analysis is based on evidence that individual synaptic activations do not reduce local driving force significantly in most central neurons, so each synapse acts approximately as a current source. Therefore the spread of PSPs throughout a dendritic tree can be described in terms of transfer impedance (Z(c)), which reflects how a current applied at one location affects membrane potential at other locations. We addressed this topic through four lines of study and uncovered new implications of neuronal morphology for synaptic integration. First, Z(c) was considered in terms of two-port theory and contrasted with dendrosomatic voltage transfer. Second, equivalent cylinder models were used to compare the spatial profiles of Z(c) and dendrosomatic voltage transfer. These simulations showed that Z(c) is less affected by dendritic location than voltage transfer is. Third, compartmental models based on morphological reconstructions of five different neuron types were used to calculate Z(c), input impedance (Z(N)), and voltage transfer throughout the dendritic tree. For all neurons, there was no significant variation of Z(c) with location within higher-order dendrites. Furthermore, Z(c) was relatively independent of synaptic location throughout the entire cell in three of the five neuron types (CA3 interneurons, CA3 pyramidal neurons, and dentate granule cells). This was quite unlike Z(N), which increased with distance from the soma and was responsible for a parallel decrease of voltage transfer. Fourth, simulations of fast excitatory PSPs (EPSPs) were consistent with the analysis of Z(c); peak EPSP amplitude varied <20% in the same three neuron types, a phenomenon that we call "passive synaptic normalization" to underscore the fact that it does not require active currents. We conclude that the presence of a long primary dendrite, as in CA1 or neocortical pyramidal cells, favors substantial location-dependent variability of somatic PSP amplitude. In neurons that lack long primary dendrites, however, PSP amplitude at the soma will be much less dependent on synaptic location.
Asunto(s)
Dendritas/fisiología , Hipocampo/fisiología , Sinapsis/fisiología , Algoritmos , Animales , Estimulación Eléctrica , Electrofisiología , Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/citología , Potenciales de la Membrana/fisiología , Modelos Neurológicos , Células Piramidales/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
1. The linear membrane responses of CA3 interneurones were determined with the use of whole-cell patch recording methods. The mean input resistance (RN) for all cells in this study was 526 +/- 16 MOmega and the slowest membrane time constant (tau0) was 73 +/- 3 ms. 2. The three-dimensional morphology of 63 biocytin-labelled neurones was used to construct compartmental models. Specific membrane resistivity (Rm) and specific membrane capacitance (Cm) were estimated by fitting the linear membrane response. Acceptable fits were obtained for 24 CA3 interneurones. The mean Rm was 61.9 +/- 34.2 Omega cm2 and the mean Cm was 0.9 +/- 0.3 microF cm-2. Intracellular resistance (Ri) could not be resolved in this study. 3. Examination of voltage attenuation revealed a significantly low synaptic efficiency from most dendritic synaptic input locations to the soma. 4. Simulations of excitatory postsynaptic potentials (EPSPs) were analysed at both the site of synaptic input and at the soma. There was little variability in the depolarization at the soma from synaptic inputs placed at different locations along the dendritic tree. The EPSP amplitude at the site of synaptic input was progressively larger with distance from the soma, consistent with a progressive increase in input impedance. 5. The 'iso-efficiency' of spatially different synaptic inputs arose from two opposing factors: an increase in EPSP amplitude at the synapse with distance from the soma was opposed by a nearly equivalent increase in voltage attenuation. These simulations suggest that, in these particular neurones, the amplitude of EPSPs measured at the soma will not be significantly affected by the location of synaptic inputs.
Asunto(s)
Hipocampo/fisiología , Interneuronas/fisiología , Análisis de Varianza , Animales , Potenciales Postsinápticos Excitadores , Técnicas In Vitro , Lisina/análogos & derivados , Potenciales de la Membrana , Modelos Neurológicos , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , SinapsisRESUMEN
In both primates and rodents, the prefrontal cortex (PFC) is highly innervated by dopaminergic fibers originating from the ventral tegmental area, and activation of this mesocortical dopaminergic system decreases spontaneous and evoked activity in the PFC in vivo. We have examined the effects of dopamine (DA), over a range of concentrations, on the passive and active membrane properties of layer V pyramidal cells from the rat medial PFC (mPFC). Whole-cell and perforated-patch recordings were made from neurons in rat mPFC. As a measure of cell excitability, trains of action potentials were evoked with 1-sec-long depolarizing current steps. Bath application of DA (0.05-30 microM) produced a reversible decrease in the number of action potentials evoked by a given current step. In addition, DA reversibly decreased the input resistance (RN) of these cells. In a subset of experiments, a transient increase in excitability was observed after the washout of DA. Control experiments suggest that these results are not attributable to changes in spontaneous synaptic activity, age-dependent processes, or strain-specific differences in dopaminergic innervation and physiology. Pharmacological analyses, using D1 agonists (SKF 38393 and SKF 81297), a D1 antagonist (SCH 23390), a D2 receptor agonist (quinpirole), and a D2 antagonist (sulpiride) suggest that decreases in spiking and RN are mediated by D2 receptor activation. Together, these results demonstrate that DA, over a range of concentrations, has an inhibitory effect on layer V pyramidal neurons in the rat mPFC, possibly through D2 receptor activation.
Asunto(s)
Dopamina/farmacología , Corteza Prefrontal/fisiología , Células Piramidales/fisiología , Potenciales de Acción/efectos de los fármacos , Factores de Edad , Animales , Animales Recién Nacidos , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Antagonistas de los Receptores de Dopamina D2 , Electrofisiología , Técnicas In Vitro , Técnicas de Placa-Clamp , Corteza Prefrontal/efectos de los fármacos , Células Piramidales/efectos de los fármacos , Ratas , Ratas Long-Evans , Ratas Sprague-Dawley , Receptores de Dopamina D1/antagonistas & inhibidores , Receptores de Dopamina D1/efectos de los fármacos , Receptores de Dopamina D1/fisiología , Receptores de Dopamina D2/efectos de los fármacos , Receptores de Dopamina D2/fisiología , TemperaturaRESUMEN
Interneurons of the hippocampal formation are traditionally identified electrophysiologically as those cells that fire trains of weakly accommodating action potentials in response to depolarizing current injection. We studied the firing properties of nonpyramidal neurons in the five substrata of the CA3b region of hippocampus. With the use of whole cell recording methods we found that nonpyramidal neurons fired in a range from weak to strong spike-frequency accommodation (SFA) that was calcium dependent. Slow afterhyperpolarizations were not associated with strong SFA. In addition a subset of interneurons ( approximately 20%) fired with an irregular firing pattern that was generally calcium independent. These results suggest a calcium-dependent mechanism for SFA in nonpyramidal neurons that is distinct from pyramidal cells and further demonstrates the heterogeneity of hippocampal interneurons.
Asunto(s)
Calcio/farmacología , Hipocampo/citología , Células Piramidales/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Calcio/metabolismo , Tamaño de la Célula/fisiología , Estimulación Eléctrica , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-DawleyRESUMEN
Confocal laser scanning microscopy was used to visualize Ca2+ transients in a particular type of dendritic spine, known as a thorny excrescence, in hippocampal CA3 pyramidal neurons. These large excrescences or thorns, which serve as the postsynaptic target for the mossy-fiber synaptic inputs, were identified on the basis of their location, frequency, and size. Whole cell recordings were made from superficial CA3 pyramidal neurons in thick hippocampal slices with the use of infrared video microscopy; cells with proximal apical dendrites close to the surface of the slice were selected. Changes in intracellular Ca2+ levels were monitored by imaging changes in fluorescence of the dyes Calcium Green-1 and Fluo-3. Dual-emission fluorescence imaging was also employed with the use of a combination of Fluo-3 and the Ca2+-insensitive dye seminaphthorhodafluor-1. This method was used to decrease the potential influence of background fluorescence on the calculated changes in intracellular Ca2+ concentration ([Ca2+]i). Somatic depolarization produced increases in [Ca2+]i in both the thorn and the immediately adjacent dendrite. Changes in [Ca2+]i were time locked with the onset of depolarization and the decay began immediately after the termination of depolarization. The peak increase in the Ca2+ signal was significantly greater in the thorns than in the adjacent dendritic shafts. With the use of high-temporal-resolution methods (line scans), differences were also seen in the time course of Ca2+ signals in these two regions. The decay time constants of the Ca2+ signal were faster in thorns than in the adjacent dendritic shafts. These observations suggest that voltage-gated Ca2+ channels are localized directly on the dendritic spines receiving mossy-fiber input. Furthermore, Ca2+ homeostasis within thorny excrescences is distinct from Ca2+ regulation in the dendritic shaft, at least over brief time periods, a finding that could have important implications for synaptic plasticity and signaling.
Asunto(s)
Calcio/metabolismo , Células Piramidales/metabolismo , Animales , Técnicas In Vitro , Potenciales de la Membrana/fisiología , Microscopía Confocal , Ratas , Ratas Sprague-Dawley , Sinapsis/fisiologíaRESUMEN
1. We tested several hypotheses with respect to the mechanisms and processes that control the firing characteristics and determine the spatial and temporal dynamics of intracellular Ca2+ in CA3 hippocampal neurons. In particular, we were interested to know 1) whether bursting and nonbursting behavior of CA3 neurons could be accounted for in a morphologically realistic model using a number of the known ionic conductances; 2) whether such a model is robust across different cell morphologies; 3) whether some particular nonuniform distribution of Ca2+ channels is required for bursting; and 4) whether such a model can reproduce the magnitude and spatial distribution of intracellular Ca2+ transients determined from fluorescence imaging studies and can predict reasonable intracellular Ca2+ concentration ([Ca2+]i) distribution for CA3 neurons. 2. For this purpose we have developed a highly detailed model of the distribution and densities of membrane ion channels in hippocampal CA3 bursting and nonbursting pyramidal neurons. This model reproduces both the experimentally observed firing modes and the dynamics of intracellular Ca2+. 3. The kinetics of the membrane ionic conductances are based on available experimental data. This model incorporates a single Na+ channel, three Ca2+ channels (CaN, CaL, and CaT), three Ca(2+)-independent K+ channels (KDR, KA, and KM), two Ca(2+)-dependent K+ channels (KC and KAHP), and intracellular Ca(2+)-related processes such as buffering, pumping, and radial diffusion. 4. To test the robustness of the model, we applied it to six different morphologically accurate reconstructions of CA3 hippocampal pyramidal neurons. In every neuron, Ca2+ channels, Ca(2+)-related processes, and Ca(2+)-dependent K+ channels were uniformly distributed over the entire cell. Ca(2+)-independent K+ channels were placed on the soma and the proximal apical dendrites. For each reconstructed cell we were able to reproduce bursting and nonbursting firing characteristics as well as Ca2+ transients and distributions for both somatic and synaptic stimulations. 5. Our simulation results suggest that CA3 pyramidal cell bursting behavior does not require any special distribution of Ca(2+)-dependent channels and mechanisms. Furthermore, a simple increase in the Ca(2+)-independent K+ conductances is sufficient to change the firing mode of our CA3 neurons from bursting to nonbursting. 6. The model also displays [Ca2+]i transients and distributions that are consistent with fluorescent imaging data. Peak [Ca2+]i distribution for synaptic stimulation of the nonbursting model is broader when compared with somatic stimulation. Somatic stimulation of the bursting model shows a broader distribution in [Ca2+]i when compared with the nonbursting model.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Simulación por Computador , Hipocampo/fisiología , Redes Neurales de la Computación , Calcio , Cinética , Matemática , Neuronas/fisiología , Sinapsis , Factores de TiempoRESUMEN
Changes in the intracellular Ca2+ concentration ([Ca2+]i) within CA1 hippocampal pyramidal neurons were imaged using confocal laser scanning microscopy in conjunction with Ca(2+)-sensitive fluorescent indicators. The imaging was performed in thick hippocampal brain slices while simultaneously measuring or controlling electrical activity with sharp microelectrodes or whole-cell patch-clamp electrodes. The combination of imaging and electrophysiology was essential for interpreting the changes in [Ca2+]i. We compared the increases in [Ca2+]i produced by either of two methods--direct depolarization of the cell via the somatic electrode or high-frequency stimulations of synaptic inputs. The increases in [Ca2+]i in the soma and proximal dendrites caused by both methods were of comparable magnitude and they always decayed within seconds in healthy cells. However, the spatial patterns of distal Ca2+ increases were different. Separate sets of synaptic inputs to the same cell resulted in different spatial patterns of [Ca2+]i transients. We isolated and observed what appeared to be a voltage-independent component of the synaptically mediated [Ca2+]i transients. This work demonstrates that the combination of neurophysiology and simultaneous confocal microscopy is well suited for visualizing and analyzing [Ca2+]i changes within highly localized regions of neurons in thick brain slices. The approach should allow further analysis of the relative contribution of voltage- and agonist-dependent influences on [Ca2+]i within neurons throughout the CNS and it raises the possibility of routinely relating subcellular [Ca2+]i changes to structural and functional modifications.
Asunto(s)
Calcio/metabolismo , Dendritas/metabolismo , Dendritas/ultraestructura , Hipocampo/metabolismo , Hipocampo/ultraestructura , Envejecimiento/metabolismo , Compuestos de Anilina , Animales , Calcio/análisis , Dendritas/química , Estimulación Eléctrica , Colorantes Fluorescentes , Hipocampo/fisiología , Procesamiento de Imagen Asistido por Computador , Masculino , Microelectrodos , Microscopía Confocal , Neuronas/fisiología , Neuronas/ultraestructura , Compuestos Orgánicos , Técnicas de Placa-Clamp , Tractos Piramidales/ultraestructura , Ratas , Ratas Sprague-Dawley , Sinapsis/fisiología , Sinapsis/ultraestructura , Grabación de Cinta de Video , XantenosRESUMEN
1. We investigated the effects of metabotropic glutamate (mGlu) receptor activation on intracellular Ca2+ concentration ([Ca2+]i) in the soma and dendrites of hippocampal CA1 pyramidal neurons. Changes in [Ca2+]i were measured using confocal imaging simultaneously with whole-cell recording techniques. Differences in [Ca2+]i were visualized as changes in the fluorescence of the Ca(2+)-sensitive dye Fluo-3. 2. Brief application of the specific mGlu receptor agonist (1S,3R)-ACPD to either the apical or basal dendrites produced initially localized increases in [Ca2+]i that subsequently propagated as waves throughout much of the neuron. These Ca2+ waves, which propagated at approximately 40 microns/s, were shown not to reflect intracellular Ca2+ diffusion or extracellular diffusion of ACPD and were always accompanied by small outward membrane currents. 3. Repetitive application of ACPD failed to trigger further Ca2+ release. We found that a threshold level of voltage-gated Ca2+ entry during trains of action potentials was needed to prime further mGlu-stimulated Ca2+ release. In contrast, the passage of time alone did not cause the mGlu-release system to reactivate--restoration of ACPD-stimulated Ca2+ release. The spike-mediated Ca2+ signal was unaffected by mGlu-stimulated depletion of intracellular stores. 4. These experiments demonstrate that specific mGlu receptor activation can mobilize Ca2+ in dendrites of CA1 neurons and trigger waves of Ca(2+)-induced Ca(2+)-release throughout the cell. A use-dependent relationship between voltage-gated Ca2+ entry during trains of action potentials and mGlu-stimulated Ca2+ release is suggested.
Asunto(s)
Canales de Calcio/fisiología , Calcio/fisiología , Dendritas/fisiología , Hipocampo/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Transmisión Sináptica/fisiología , Animales , Técnicas de Cultivo , Potenciales de la Membrana/fisiología , Células Piramidales/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
The dendritic trees of neurons are structurally and functionally complex integrative units receiving thousands of synaptic inputs that have excitatory and inhibitory, fast and slow, and electrical and biochemical effects. The pattern of activation of these synaptic inputs determines if the neuron will fire an action potential at any given point in time and how it will respond to similar inputs in the future. Two critical factors affect the integrative function of dendrites: the distribution of voltage-gated ion channels in the dendritic tree and the passive electrical properties, or 'electrotonic structure', upon which these active channels are superimposed. The authors review recent data from patch-clamp recordings that provide new estimates of the passive membrane properties of hippocampal neurons, and show, with examples, how these properties affect the shaping and attenuation of synaptic potentials as they propagate in the dendrites, as well as how they affect the measurement of current from synapses located in the dendrites. Voltage-gated channels might influence the measurement of 'passive' membrane properties and, reciprocally, passive membrane properties might affect the activation of voltage-gated channels in dendrites.
Asunto(s)
Membrana Celular/fisiología , Dendritas/fisiología , Transmisión Sináptica/fisiología , Animales , Hipocampo/citología , Activación del Canal Iónico , Neuronas/fisiología , Neuronas/ultraestructura , RatasRESUMEN
The induction of long-term potentiation (LTP) is generally assumed to be triggered by Ca2+ entry into dendritic spines via NMDA receptor-gated channels. A previous computational model proposed that spines serve several functions in this process. First, they compartmentalize and amplify increase in [Ca2+]i. Second, they augment the nonlinear relationship between synaptic strength and the probability or magnitude of LTP induction. Third, they isolate the metabolic machinery responsible for LTP induction from increases in [Ca2+]i produced by voltage-gated Ca2+ channels in the dendritic shaft. Here we examine this last prediction of the model using methods that combine confocal microscopy with simultaneous neurophysiological recordings in hippocampal brain slices. Either of two Ca(2+)-sensitive dyes were injected into CA1 pyramidal neurons. Direct depolarization of the neurons via the somatic electrode produced clear increases in Ca2+ signals within the dendritic spines, a result that was not predicted by the previous spine model. Our new spine model suggests that some of this signal could theoretically result from Ca(2+)-bound dye diffusing from the dendritic shaft into the spine. Dye diffusion alone cannot, however, explain the numerous cases in which the Ca2+ signal in the spine was considerably larger than that in the adjacent dendritic shaft. The latter observations raise the possibility of voltage-gated Ca2+ entry directly into the spine or else perhaps via Ca(2+)-dependent Ca2+ release. The new spine model accommodates these observations as well as several other recent experimental results.
Asunto(s)
Canales de Calcio/fisiología , Calcio/fisiología , Potenciación a Largo Plazo , Microscopía Fluorescente/métodos , Células Piramidales/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Dendritas/fisiología , Activación del Canal Iónico , Rayos Láser , Microscopía Fluorescente/instrumentación , Grabación en VideoRESUMEN
1. High-speed fluorescence imaging was used to measure intracellular Ca2+ concentration ([Ca2+]i) changes in hippocampal neurons injected with the Ca(2+)-sensitive indicator fura-2 during intrasomatic and synaptic stimulation. The results of these experiments were used to construct a biophysical model of [Ca2+]i dynamics in hippocampal neurons. 2. A compartmental model of a pyramidal neuron was constructed incorporating published passive membrane properties of these cells, three types of voltage-gated Ca2+ channels characterized from adult hippocampal neurons, voltage-gated Na+ and K+ currents, and mechanisms for Ca2+ buffering and extrusion. 3. In hippocampal pyramidal neurons imaging of Na+ entry during electrical activity suggests that Na+ channels, at least in sufficient density to sustain action potentials, are localized in the soma and the proximal part of the apical dendritic tree. The model, which incorporates this distribution, demonstrates that action potentials attenuate steeply in passive distal dendritic compartments or distal dendritic compartments containing Ca2+ and K+ channels. This attenuation was affected by intracellular resistivity but not membrane resistivity. 4. Consistent with fluorescence imaging experiments, a non-uniform distribution of Ca2+ accumulation was generated by Ca2+ entry through voltage-gated Ca2+ channels opened by decrementally propagating Na+ action potentials. Consequently, the largest increases in [C2+]i were produced in the proximal dendrites. Distal voltage-gated Ca2+ currents were activated by broad, almost isopotential action potentials produced by reducing the overall density of K+ channels. 5. Simulations of subthreshold synaptic stimulation produced dendritic Ca2+ entry by the activation of voltage-gated Ca2+ channels. In the model these Ca2+ signals were localized near the site of synaptic input because of the attenuation of synaptic potentials with distance from the site of origin and the steep voltage-dependence of Ca2+ channel activation. 6. These simulations support the hypotheses generated from experimental evidence regarding the differential distribution of voltage-gated Ca2+ and Na+ channels in hippocampal neurons and the resulting voltage-gated Ca2+ accumulation from action and synaptic potentials.
Asunto(s)
Canales de Calcio/fisiología , Calcio/metabolismo , Simulación por Computador , Dendritas/fisiología , Hipocampo/fisiología , Modelos Neurológicos , Transmisión Sináptica/fisiología , Animales , Técnicas de Cultivo , Potenciales de la Membrana/fisiología , Microscopía Fluorescente , Neuronas/fisiología , Canales de Potasio/fisiología , Ratas , Canales de Sodio/fisiologíaRESUMEN
1. The voltage- and space-clamp errors associated with the use of a somatic electrode to measure current from dendritic synapses are evaluated using both equivalent-cylinder and morphologically realistic models of neuronal dendritic trees. 2. As a first step toward understanding the properties of synaptic current distortion under voltage-clamp conditions, the attenuation of step and sinusoidal voltage changes are evaluated in equivalent cylinder models. Demonstration of the frequency-dependent attenuation of voltage in the cable is then used as a framework for understanding the distortion of synaptic currents generated at sites remote from the somatic recording electrode and measured in the voltage-clamp recording configuration. 3. Increases in specific membrane resistivity (Rm) are shown to reduce steady-state voltage attenuation, while producing only minimal reduction in attenuation of transient voltage changes. Experimental manipulations that increase Rm therefore improve the accuracy of estimates of reversal potential for electrotonically remote synapses, but do not significantly reduce the attenuation of peak current. In addition, increases in Rm have the effect of slowing the kinetics of poorly clamped synaptic currents. 4. The effects of the magnitude of the synaptic conductance and its kinetics on the measured synaptic currents are also examined and discussed. The error in estimating parameters from measured synaptic currents is greatest for synapses with fast kinetics and large conductances. 5. A morphologically realistic model of a CA3 pyramidal neuron is used to demonstrate the generality of the conclusions derived from equivalent cylinder models. The realistic model is also used to fit synaptic currents generated by stimulation of mossy fiber (MF) and commissural/associational (C/A) inputs to CA3 neurons and to estimate the amount of distortion of these measured currents. 6. Anatomic data from the CA3 pyramidal neuron model are used to construct a simplified two-cylinder CA3 model. This model is used to estimate the electrotonic distances of MF synapses (which are located proximal to the soma) and perforant path (PP) synapses (which are located at the distal ends of the apical dendrites) and the distortion of synaptic current parameters measured for these synapses. 7. Results from the equivalent-cylinder models, the morphological CA3 model, and the simplified CA3 model all indicate that the amount of distortion of synaptic currents increases steeply as a function of distance from the soma. MF synapses close to the soma are likely to be subject only to small space-clamp errors, whereas MF synapses farther from the soma are likely to be substantially attenuated.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Dendritas/fisiología , Hipocampo/fisiología , Sinapsis/fisiología , Transmisión Sináptica/fisiología , Animales , Cobayas , Potenciales de la Membrana/fisiología , Modelos Neurológicos , Conducción Nerviosa/fisiología , Neuronas/fisiologíaRESUMEN
The dendrites of many types of neurons contain voltage-dependent Na+ and Ca2+ conductances that generate action potentials (see ref. 1 for review). The function of these spikes is not well understood, but the Ca2+ entry stimulated by spikes probably affects Ca(2+)-dependent processes in dendrites. These include synaptic plasticity, cytotoxicity and exocytosis. Several lines of evidence suggest that dendritic spikes occur within subregions of the dendrites. To study the mechanism that govern the spread of spikes in the dendrites of hippocampal pyramidal cells, we imaged Ca2+ entry with Fura-2 (ref. 9) and Na+ entry with a newly developed Na(+)-sensitive dye. Our results indicate that Ca2+ entry into dendrites is triggered by Na+ spikes that actively invade the dendrites. The restricted spatial distribution of Ca2+ entry seems to depend on the spread of Na+ spikes in the dendrites, rather than on a limited distribution of Ca2+ channels. In addition, we have observed an activity-dependent process that modulates the invasion of spikes into the dendrites and progressively restricts Ca2+ entry to more proximal dendritic regions.
Asunto(s)
Calcio/metabolismo , Dendritas/fisiología , Hipocampo/fisiología , Neuronas/fisiología , Tractos Piramidales/fisiología , Sodio/metabolismo , 6-Ciano 7-nitroquinoxalina 2,3-diona , Animales , Benzofuranos , Dendritas/metabolismo , Estimulación Eléctrica , Éteres Cíclicos , Colorantes Fluorescentes , Fura-2 , Hipocampo/metabolismo , Técnicas In Vitro , Masculino , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Tractos Piramidales/metabolismo , Quinoxalinas/farmacología , Ratas , Ratas Endogámicas , Espectrometría de Fluorescencia , Tetraetilamonio , Compuestos de Tetraetilamonio/farmacología , Tetrodotoxina/farmacologíaRESUMEN
We investigated the relationship between the effects of cholecystokinin (CCK) and kainic acid (KA) in the CA3 region of hippocampal slices from rats. As has been reported previously, KA in nanomolar concentrations caused spontaneous epileptiform discharges (bursts) and an excitatory shift of the input/output (I/O) curve. CCK octapeptide (100-200 nM) applied alone had no effect on spontaneous activity or I/O curves. Pretreatment of slices with sulfated CCK blocked the effect of KA on synaptic transmission, but had no effect on KA-induced bursting. Pretreatment with nonsulfated CCK had no effect.