RESUMEN
Nociceptive sensory neurons convey pain-related signals to the CNS using action potentials. Loss-of-function mutations in the voltage-gated sodium channel NaV1.7 cause insensitivity to pain (presumably by reducing nociceptor excitability) but clinical trials seeking to treat pain by inhibiting NaV1.7 pharmacologically have struggled. This may reflect the variable contribution of NaV1.7 to nociceptor excitability. Contrary to claims that NaV1.7 is necessary for nociceptors to initiate action potentials, we show that nociceptors can achieve similar excitability using different combinations of NaV1.3, NaV1.7, and NaV1.8. Selectively blocking one of those NaV subtypes reduces nociceptor excitability only if the other subtypes are weakly expressed. For example, excitability relies on NaV1.8 in acutely dissociated nociceptors but responsibility shifts to NaV1.7 and NaV1.3 by the fourth day in culture. A similar shift in NaV dependence occurs in vivo after inflammation, impacting ability of the NaV1.7-selective inhibitor PF-05089771 to reduce pain in behavioral tests. Flexible use of different NaV subtypes exemplifies degeneracy - achieving similar function using different components - and compromises reliable modulation of nociceptor excitability by subtype-selective inhibitors. Identifying the dominant NaV subtype to predict drug efficacy is not trivial. Degeneracy at the cellular level must be considered when choosing drug targets at the molecular level.
Asunto(s)
Analgésicos , Bencenosulfonamidas , Nociceptores , Éteres Fenílicos , Animales , Analgésicos/farmacología , Nociceptores/metabolismo , Nociceptores/efectos de los fármacos , Canal de Sodio Activado por Voltaje NAV1.7/metabolismo , Canal de Sodio Activado por Voltaje NAV1.7/genética , Ratones , Potenciales de Acción/efectos de los fármacos , Dolor/tratamiento farmacológico , Humanos , Canales de Sodio/metabolismo , Canales de Sodio/genética , Canal de Sodio Activado por Voltaje NAV1.8/metabolismo , Canal de Sodio Activado por Voltaje NAV1.8/genéticaRESUMEN
Electrically activating mechanoreceptive afferents inhibits pain. However, paresthesia evoked by spinal cord stimulation (SCS) at 40-60 Hz becomes uncomfortable at high pulse amplitudes, limiting SCS "dosage." Kilohertz-frequency SCS produces analgesia without paresthesia and is thought, therefore, not to activate afferent axons. We show that paresthesia is absent not because axons do not spike but because they spike asynchronously. In a pain patient, selectively increasing SCS frequency abolished paresthesia and epidurally recorded evoked compound action potentials (ECAPs). Dependence of ECAP amplitude on SCS frequency was reproduced in pigs, rats, and computer simulations and is explained by overdrive desynchronization: spikes desychronize when axons are stimulated faster than their refractory period. Unlike synchronous spikes, asynchronous spikes fail to produce paresthesia because their transmission to somatosensory cortex is blocked by feedforward inhibition. Our results demonstrate how stimulation frequency impacts synchrony based on axon properties and how synchrony impacts sensation based on circuit properties.
Asunto(s)
Estimulación de la Médula Espinal , Médula Espinal , Humanos , Ratas , Animales , Porcinos , Médula Espinal/fisiología , Estimulación de la Médula Espinal/métodos , Parestesia , Estimulación Eléctrica , Sensación , DolorRESUMEN
Neurons can use different aspects of their spiking to simultaneously represent (multiplex) different features of a stimulus. For example, some pyramidal neurons in primary somatosensory cortex (S1) use the rate and timing of their spikes to, respectively, encode the intensity and frequency of vibrotactile stimuli. Doing so has several requirements. Because they fire at low rates, pyramidal neurons cannot entrain 1:1 with high-frequency (100 to 600 Hz) inputs and, instead, must skip (i.e., not respond to) some stimulus cycles. The proportion of skipped cycles must vary inversely with stimulus intensity for firing rate to encode stimulus intensity. Spikes must phase-lock to the stimulus for spike times (intervals) to encode stimulus frequency, but, in addition, skipping must occur irregularly to avoid aliasing. Using simulations and in vitro experiments in which mouse S1 pyramidal neurons were stimulated with inputs emulating those induced by vibrotactile stimuli, we show that fewer cycles are skipped as stimulus intensity increases, as required for rate coding, and that intrinsic or synaptic noise can induce irregular skipping without disrupting phase locking, as required for temporal coding. This occurs because noise can modulate the reliability without disrupting the precision of spikes evoked by small-amplitude, fast-onset signals. Specifically, in the fluctuation-driven regime associated with sparse spiking, rate and temporal coding are both paradoxically improved by the strong synaptic noise characteristic of the intact cortex. Our results demonstrate that multiplexed coding by S1 pyramidal neurons is not only feasible under in vivo conditions, but that background synaptic noise is actually beneficial.
Asunto(s)
Ruido , Células Piramidales , Corteza Somatosensorial , Tacto , Potenciales de Acción/fisiología , Animales , Ratones , Células Piramidales/fisiología , Reproducibilidad de los Resultados , Corteza Somatosensorial/fisiología , Tacto/fisiología , VibraciónRESUMEN
Neurons regulate their excitability by adjusting their ion channel levels. Degeneracy - achieving equivalent outcomes (excitability) using different solutions (channel combinations) - facilitates this regulation by enabling a disruptive change in one channel to be offset by compensatory changes in other channels. But neurons must coregulate many properties. Pleiotropy - the impact of one channel on more than one property - complicates regulation because a compensatory ion channel change that restores one property to its target value often disrupts other properties. How then does a neuron simultaneously regulate multiple properties? Here, we demonstrate that of the many channel combinations producing the target value for one property (the single-output solution set), few combinations produce the target value for other properties. Combinations producing the target value for two or more properties (the multioutput solution set) correspond to the intersection between single-output solution sets. Properties can be effectively coregulated only if the number of adjustable channels (nin) exceeds the number of regulated properties (nout). Ion channel correlations emerge during homeostatic regulation when the dimensionality of solution space (nin - nout) is low. Even if each property can be regulated to its target value when considered in isolation, regulation as a whole fails if single-output solution sets do not intersect. Our results also highlight that ion channels must be coadjusted with different ratios to regulate different properties, which suggests that each error signal drives modulatory changes independently, despite those changes ultimately affecting the same ion channels.
Asunto(s)
Canales Iónicos , Neuronas , Homeostasis , Canales Iónicos/fisiología , Neuronas/fisiologíaRESUMEN
Neuropathic pain is a debilitating condition caused by the abnormal processing of somatosensory input. Synaptic inhibition in the spinal dorsal horn plays a key role in that processing. Mechanical allodynia - the misperception of light touch as painful - occurs when inhibition is compromised. Disinhibition is due primarily to chloride dysregulation caused by hypofunction of the potassium-chloride co-transporter KCC2. Here we show, in rats, that excitatory neurons are disproportionately affected. This is not because chloride is differentially dysregulated in excitatory and inhibitory neurons, but, rather, because excitatory neurons rely more heavily on inhibition to counterbalance strong excitation. Receptive fields in both cell types have a center-surround organization but disinhibition unmasks more excitatory input to excitatory neurons. Differences in intrinsic excitability also affect how chloride dysregulation affects spiking. These results deepen understanding of how excitation and inhibition are normally balanced in the spinal dorsal horn, and how their imbalance disrupts somatosensory processing.
Asunto(s)
Cloruros/metabolismo , Neuronas/metabolismo , Asta Dorsal de la Médula Espinal/metabolismo , Animales , Femenino , Hiperalgesia/metabolismo , Masculino , Modelos Animales , Modelos Biológicos , Fenómenos Fisiológicos del Sistema Nervioso , Neuralgia/metabolismo , Ratas , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Simportadores/metabolismo , Cotransportadores de K ClRESUMEN
Multiplexing refers to the simultaneous encoding of two or more signals. Neurons have been shown to multiplex, but different stimuli require different multiplexing strategies. Whereas the frequency and amplitude of periodic stimuli can be encoded by the timing and rate of the same spikes, natural scenes, which comprise areas over which intensity varies gradually and sparse edges where intensity changes abruptly, require a different multiplexing strategy. Recording in vivo from neurons in primary somatosensory cortex during tactile stimulation, we found that stimulus onset and offset (edges) evoked highly synchronized spiking, whereas other spikes in the same neurons occurred asynchronously. Stimulus intensity modulated the rate of asynchronous spiking, but did not affect the timing of synchronous spikes. From this, we hypothesized that spikes driven by high- and low-contrast stimulus features can be distinguished on the basis of their synchronization, and that differentially synchronized spiking can thus be used to form multiplexed representations. Applying a Bayesian decoding method, we verified that information about high- and low-contrast features can be recovered from an ensemble of model neurons receiving common input. Equally good decoding was achieved by distinguishing synchronous from asynchronous spikes and applying reverse correlation methods separately to each spike type. This result, which we verified with patch clamp recordings in vitro, demonstrates that neurons receiving common input can use the rate of asynchronous spiking to encode the intensity of low-contrast features while using the timing of synchronous spikes to encode the occurrence of high-contrast features. We refer to this strategy as synchrony-division multiplexing.
Asunto(s)
Neuronas/fisiología , Corteza Somatosensorial/fisiología , Animales , RatonesRESUMEN
Characterizing the cellular targets of kHz (1-10 kHz) electrical stimulation remains a pressing topic in neuromodulation because expanding interest in clinical application of kHz stimulation has surpassed mechanistic understanding. The presumed cellular targets of brain stimulation do not respond to kHz frequencies according to conventional electrophysiology theory. Specifically, the low-pass characteristics of cell membranes are predicted to render kHz stimulation inert, especially given the use of limited-duty-cycle biphasic pulses. Precisely because kHz frequencies are considered supra-physiological, conventional instruments designed for neurophysiological studies such as stimulators, amplifiers and recording microelectrodes do not operate reliably at these high rates. Moreover, for pulsed waveforms, the signal frequency content is well above the pulse repetition rate. Thus, the very tools used to characterize the effects of kHz electrical stimulation may themselves be confounding factors. We illustrate custom equipment design that supports reliable electrophysiological recording during kHz-rate stimulation. Given the increased importance of kHz stimulation in clinical domains and compelling possibilities that mechanisms of actions may reflect yet undiscovered neurophysiological phenomena, attention to suitable performance of electrophysiological equipment is pivotal.
Asunto(s)
Estimulación Eléctrica/instrumentación , Fenómenos Electrofisiológicos , MicroelectrodosRESUMEN
Continuation of spiking after a stimulus ends (i.e. persistent spiking) is thought to support working memory. Muscarinic receptor activation enables persistent spiking among synaptically isolated pyramidal neurons in anterior cingulate cortex (ACC), but a detailed characterization of that spiking is lacking and the underlying mechanisms remain unclear. Here, we show that the rate of persistent spiking in ACC neurons is insensitive to the intensity and number of triggers, but can be modulated by injected current, and that persistent spiking can resume after several seconds of hyperpolarization-imposed quiescence. Using electrophysiology and calcium imaging in brain slices from male rats, we determined that canonical transient receptor potential (TRPC) channels are necessary for persistent spiking and that TRPC-activating calcium enters in a spike-dependent manner via voltage-gated calcium channels. Constrained by these biophysical details, we built a computational model that reproduced the observed pattern of persistent spiking. Nonlinear dynamical analysis of that model revealed that TRPC channels become fully activated by the small rise in intracellular calcium caused by evoked spikes. Calcium continues to rise during persistent spiking, but because TRPC channel activation saturates, firing rate stabilizes. By calcium rising higher than required for maximal TRPC channel activation, TRPC channels are able to remain active during periods of hyperpolarization-imposed quiescence (until calcium drops below saturating levels) such that persistent spiking can resume when hyperpolarization is discontinued. Our results thus reveal that the robust intrinsic bistability exhibited by ACC neurons emerges from the nonlinear positive feedback relationship between spike-dependent calcium influx and TRPC channel activation.SIGNIFICANCE STATEMENT Neurons use action potentials, or spikes, to encode information. Some neurons can store information for short periods (seconds to minutes) by continuing to spike after a stimulus ends, thus enabling working memory. This so-called "persistent" spiking occurs in many brain areas and has been linked to activation of canonical transient receptor potential (TRPC) channels. However, TRPC activation alone is insufficient to explain many aspects of persistent spiking such as resumption of spiking after periods of imposed quiescence. Using experiments and simulations, we show that calcium influx caused by spiking is necessary and sufficient to activate TRPC channels and that the ensuing positive feedback interaction between intracellular calcium and TRPC channel activation can account for many hitherto unexplained aspects of persistent spiking.
Asunto(s)
Señalización del Calcio/fisiología , Giro del Cíngulo/citología , Giro del Cíngulo/fisiología , Neuronas/fisiología , Canales de Potencial de Receptor Transitorio/fisiología , Algoritmos , Animales , Canales de Calcio/fisiología , Simulación por Computador , Fenómenos Electrofisiológicos/fisiología , Retroalimentación Psicológica , Masculino , Dinámicas no Lineales , Técnicas de Placa-Clamp , Células Piramidales/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Kilohertz-frequency electric field stimulation (kEFS) applied to the spinal cord can reduce chronic pain without causing the buzzing sensation (paresthesia) associated with activation of dorsal column fibers. This suggests that high-rate spinal cord stimulation (SCS) has a mode of action distinct from conventional, parasthesia-based SCS. A recent study reported that kEFS hyperpolarizes spinal neurons, yet this potentially transformative mode of action contradicts previous evidence that kEFS induces depolarization and was based on patch clamp recordings whose accuracy in the presence of kEFS has not been verified. OBJECTIVES: We sought to elucidate the basis for kEFS-induced hyperpolarization and to validate the effects of kEFS observed in patch clamp recordings by comparing with independent optical methods. METHODS: Using patch clamp electrophysiology and voltage-sensitive dye (VSD) imaging, we measured the response to kEFS applied in vitro to hippocampal and spinal neurons. RESULTS: The kEFS-induced hyperpolarization observed with current clamp recordings was corroborated by VSD imaging and rheobase measurements in patched neurons. However, no hyperpolarization was observed when imaging unpatched neurons or when recording with a voltage-follower amplifier rather than with a patch clamp amplifier (PCA). We found that EFS induced an artifactual current in PCAs that was injected back into current clamped neurons. The artifactual current induced by single, charge-balanced EFS pulses caused modest hyperpolarization, but these unitary hyperpolarizations accumulated when EFS pulses were repeated at kilohertz frequencies. CONCLUSION: Our results rule out hyperpolarization as the mechanism underlying kEFS-mediated analgesia and highlight the risk of recording artifacts caused by extracellular electrical stimulation.
Asunto(s)
Artefactos , Estimulación Eléctrica/métodos , Hipocampo/citología , Neuronas/fisiología , Médula Espinal/citología , Animales , Células Cultivadas , Técnicas de Placa-Clamp , Ratas , Imagen de Colorante Sensible al VoltajeRESUMEN
The cortex encodes a broad range of inputs. This breadth of operation requires sensitivity to weak inputs yet non-saturating responses to strong inputs. If individual pyramidal neurons were to have a narrow dynamic range, as previously claimed, then staggered all-or-none recruitment of those neurons would be necessary for the population to achieve a broad dynamic range. Contrary to this explanation, we show here through dynamic clamp experiments in vitro and computer simulations that pyramidal neurons have a broad dynamic range under the noisy conditions that exist in the intact brain due to background synaptic input. Feedforward inhibition capitalizes on those noise effects to control neuronal gain and thereby regulates the population dynamic range. Importantly, noise allows neurons to be recruited gradually and occludes the staggered recruitment previously attributed to heterogeneous excitation. Feedforward inhibition protects spike timing against the disruptive effects of noise, meaning noise can enable the gain control required for rate coding without compromising the precise spike timing required for temporal coding.
Asunto(s)
Corteza Cerebral/fisiología , Inhibición Neural/fisiología , Células Piramidales/fisiología , Sinapsis/fisiología , Transmisión Sináptica/fisiología , Animales , Simulación por Computador , Masculino , Modelos Neurológicos , Técnicas de Placa-Clamp , Ratas Sprague-Dawley , Factores de Tiempo , Técnicas de Cultivo de TejidosRESUMEN
Neuropathic pain, which arises from damage to the nervous system, is a major unmet clinical challenge. Reversing the neuronal hyperexcitability induced by nerve damage is a logical treatment strategy but has proven frustratingly difficult. Here, we propose a novel explanation for that difficulty. Changes in several different ion channels are individually sufficient to cause hyperexcitability in primary somatosensory neurons. Despite offering multiple drug targets, this scenario is problematic: if multiple sufficient changes are triggered by nerve injury, then no single change is necessary for hyperexcitability. This so-called degeneracy compromises therapeutic interventions because drug effects on any one ion channel can be circumvented by changes occurring in other ion channels. Overcoming degeneracy demands a more integrative approach to drug discovery.
Asunto(s)
Potenciales de Acción/fisiología , Vías Aferentes/fisiopatología , Dolor Crónico/fisiopatología , Hiperalgesia/fisiopatología , Canales Iónicos/metabolismo , Neuralgia/fisiopatología , Neuronas Aferentes/metabolismo , Vías Aferentes/metabolismo , Dolor Crónico/metabolismo , Humanos , Hiperalgesia/metabolismo , Neuralgia/metabolismo , Neuronas/metabolismo , Neuronas/fisiología , Neuronas Aferentes/fisiologíaRESUMEN
Neuropathic pain remains notoriously difficult to treat despite numerous drug targets. Here, we offer a novel explanation for this intractability. Computer simulations predicted that qualitative changes in primary afferent excitability linked to neuropathic pain arise through a switch in spike initiation dynamics when molecular pathologies reach a tipping point (criticality), and that this tipping point can be reached via several different molecular pathologies (degeneracy). We experimentally tested these predictions by pharmacologically blocking native conductances and/or electrophysiologically inserting virtual conductances. Multiple different manipulations successfully reproduced or reversed neuropathic changes in primary afferents from naïve or nerve-injured rats, respectively, thus confirming the predicted criticality and its degenerate basis. Degeneracy means that several different molecular pathologies are individually sufficient to cause hyperexcitability, and because several such pathologies co-occur after nerve injury, that no single pathology is uniquely necessary. Consequently, single-target-drugs can be circumvented by maladaptive plasticity in any one of several ion channels. DOI: http://dx.doi.org/10.7554/eLife.02370.001.
Asunto(s)
Ganglios Espinales/lesiones , Ganglios Espinales/fisiopatología , Neuralgia/fisiopatología , Animales , Simulación por Computador , Modelos Animales de Enfermedad , Electrofisiología , Ganglios Espinales/citología , Masculino , Modelos Moleculares , Modelos Neurológicos , Dinámicas no Lineales , Ratas , Ratas Sprague-DawleyRESUMEN
Neurons rely on action potentials, or spikes, to encode information. But spikes can encode different stimulus features in different neurons. We show here through simulations and experiments how neurons encode the integral or derivative of their input based on the distinct tuning properties conferred upon them by subthreshold currents. Slow-activating subthreshold inward (depolarizing) current mediates positive feedback control of subthreshold voltage, sustaining depolarization and allowing the neuron to spike on the basis of its integrated stimulus waveform. Slow-activating subthreshold outward (hyperpolarizing) current mediates negative feedback control of subthreshold voltage, truncating depolarization and forcing the neuron to spike on the basis of its differentiated stimulus waveform. Depending on its direction, slow-activating subthreshold current cooperates or competes with fast-activating inward current during spike initiation. This explanation predicts that sensitivity to the rate of change of stimulus intensity differs qualitatively between integrators and differentiators. This was confirmed experimentally in spinal sensory neurons that naturally behave as specialized integrators or differentiators. Predicted sensitivity to different stimulus features was confirmed by covariance analysis. Integration and differentiation, which are themselves inverse operations, are thus shown to be implemented by the slow feedback mediated by oppositely directed subthreshold currents expressed in different neurons.
RESUMEN
Neural networks are more than the sum of their parts, but the properties of those parts are nonetheless important. For instance, neuronal properties affect the degree to which neurons receiving common input will spike synchronously, and whether that synchrony will propagate through the network. Stimulus-evoked synchrony can help or hinder network coding depending on the type of code. In this Perspective, we describe how spike initiation dynamics influence neuronal input-output properties, how those properties affect synchronization, and how synchronization affects network coding. We propose that synchronous and asynchronous spiking can be used to multiplex temporal (synchrony) and rate coding and discuss how pyramidal neurons would be well suited for that task.
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Potenciales de Acción/fisiología , Modelos Neurológicos , Red Nerviosa/fisiología , Neuronas/fisiología , Dinámicas no Lineales , AnimalesRESUMEN
Pain, itch, heat, cold, and touch represent different percepts arising from somatosensory input. How stimuli give rise to these percepts has been debated for over a century. Recent work supports the view that primary afferents are highly specialized to transduce and encode specific stimulus modalities. However, cross-modal interactions (e.g. inhibition or exacerbation of pain by touch) support convergence rather than specificity in central circuits. We outline how peripheral specialization together with central convergence could enable spinal microcircuits to combine inputs from distinctly specialized, co-activated afferents and to modulate the output signals thus formed through computations like normalization. These issues will be discussed alongside recent advances in our understanding of microcircuitry in the superficial dorsal horn.
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Vías Aferentes/patología , Red Nerviosa/fisiopatología , Dolor/patología , Médula Espinal/patología , Potenciales de Acción , Animales , Humanos , Red Nerviosa/patología , Neuronas/citología , Neuronas/fisiologíaRESUMEN
Correlated spiking has been widely observed, but its impact on neural coding remains controversial. Correlation arising from comodulation of rates across neurons has been shown to vary with the firing rates of individual neurons. This translates into rate and correlation being equivalently tuned to the stimulus; under those conditions, correlated spiking does not provide information beyond that already available from individual neuron firing rates. Such correlations are irrelevant and can reduce coding efficiency by introducing redundancy. Using simulations and experiments in rat hippocampal neurons, we show here that pairs of neurons receiving correlated input also exhibit correlations arising from precise spike-time synchronization. Contrary to rate comodulation, spike-time synchronization is unaffected by firing rate, thus enabling synchrony- and rate-based coding to operate independently. The type of output correlation depends on whether intrinsic neuron properties promote integration or coincidence detection: "ideal" integrators (with spike generation sensitive to stimulus mean) exhibit rate comodulation, whereas ideal coincidence detectors (with spike generation sensitive to stimulus variance) exhibit precise spike-time synchronization. Pyramidal neurons are sensitive to both stimulus mean and variance, and thus exhibit both types of output correlation proportioned according to which operating mode is dominant. Our results explain how different types of correlations arise based on how individual neurons generate spikes, and why spike-time synchronization and rate comodulation can encode different stimulus properties. Our results also highlight the importance of neuronal properties for population-level coding insofar as neural networks can employ different coding schemes depending on the dominant operating mode of their constituent neurons.
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Potenciales de Acción/fisiología , Modelos Neurológicos , Neuronas/fisiología , Animales , Masculino , Distribución Normal , Técnicas de Cultivo de Órganos , Ratas , Ratas Sprague-DawleyRESUMEN
Synaptic inhibition by GABA(A) receptors requires a transmembrane chloride gradient. Hyperpolarization or shunting results from outward current produced by chloride flowing down this gradient, into the cell. Chloride influx necessarily depletes the chloride gradient. Therefore, mechanisms that replenish the gradient (by reducing intracellular chloride concentration, [Cl(-)](i)) are crucial for maintaining the efficacy of GABA(A) receptor-mediated inhibition. ClC-2 is an inward-rectifying chloride channel that is thought to help extrude chloride because inward rectification should, in principle, allow ClC-2 to act as a one-way chloride exit valve. But chloride efflux via ClC-2 nevertheless requires an appropriate driving force. Using computer modeling, we reproduced voltage-clamp experiments showing chloride efflux via ClC-2, but testing the same model under physiological conditions revealed that ClC-2 normally leaks chloride into the cell. The discrepancy is explained by the driving force conditions that exist under artificial versus physiological conditions, and by the fact that ClC-2 rectification is neither complete nor instantaneous. Thus, contrary to previous assertions that ClC-2 helps maintain synaptic inhibition by lowering [Cl(-)](i), our simulations show that ClC-2 mediates chloride influx, thus producing outward current and directly reducing excitability. To test how ClC-2 functions in real neurons, we used dynamic clamp to insert virtual ClC-2 channels into rat CA1 pyramidal cells with and without native ClC-2 channels blocked. Experiments confirmed that ClC-2 reduces spiking independently of inhibitory synaptic transmission. Our results highlight the importance of considering driving force when inferring how a channel functions under physiological conditions.
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Fenómenos Biofísicos/fisiología , Canales de Cloruro/fisiología , Cloruros/metabolismo , Líquido Intracelular/metabolismo , Neuronas/fisiología , Animales , Simulación por Computador , Estimulación Eléctrica , Potenciales de la Membrana/fisiología , Modelos Neurológicos , Técnicas de Placa-Clamp , Ratas , Receptores de GABA-A/fisiología , Simportadores/metabolismo , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/metabolismo , Cotransportadores de K ClRESUMEN
During wakefulness, pyramidal neurons in the intact brain are bombarded by synaptic input that causes tonic depolarization, increased membrane conductance (i.e., shunting), and noisy fluctuations in membrane potential; by comparison, pyramidal neurons in acute slices typically experience little background input. Such differences in operating conditions can compromise extrapolation of in vitro data to explain neuronal operation in vivo. For instance, pyramidal neurons have been identified as integrators (i.e., class 1 neurons according to Hodgkin's classification of intrinsic excitability) based on in vitro experiments but that classification is inconsistent with the ability of hippocampal pyramidal neurons to oscillate/resonate at theta frequency since intrinsic oscillatory behavior is limited to class 2 neurons. Using long depolarizing stimuli and dynamic clamp to reproduce in vivo-like conditions in slice experiments, we show that CA1 hippocampal pyramidal cells switch from integrators to resonators, i.e., from class 1 to class 2 excitability. The switch is explained by increased outward current contributed by the M-type potassium current I(M), which shifts the balance of inward and outward currents active at perithreshold potentials and thereby converts the spike-initiating mechanism as predicted by dynamical analysis of our computational model. Perithreshold activation of I(M) is enhanced by the depolarizing shift in spike threshold caused by shunting and/or sodium channel inactivation secondary to tonic depolarization. Our conclusions were validated by multiple comparisons between simulation and experimental data. Thus even so-called "intrinsic" properties may differ qualitatively between in vitro and in vivo conditions.
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Potenciales de la Membrana/fisiología , Células Piramidales/fisiología , 6-Ciano 7-nitroquinoxalina 2,3-diona/farmacología , Animales , Bicuculina/farmacología , Biofisica , Simulación por Computador , Umbral Diferencial/fisiología , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Antagonistas del GABA/farmacología , Hipocampo/citología , Técnicas In Vitro , Masculino , Potenciales de la Membrana/efectos de los fármacos , Modelos Neurológicos , Dinámicas no Lineales , Técnicas de Placa-Clamp/métodos , Células Piramidales/efectos de los fármacos , Ratas , Ratas Sprague-DawleyRESUMEN
Prion diseases are heterogeneous in clinical presentation, suggesting that different prion diseases have distinct pathophysiological changes. To understand the pathophysiology specific to variant Creutzfeldt-Jakob Disease (vCJD), in vitro electrophysiological studies were performed in a mouse model in which human-derived vCJD prions were transmitted to transgenic mice expressing human instead of murine prion protein. Paired-pulse stimulation of the Schaffer collaterals evoked hypersynchronous bursting in the hippocampus of vCJD-inoculated mice; comparable bursts were never observed in control or Prnp knockout mice, or in mice inoculated with a strain of prion associated with classical CJD. Furthermore, NMDA receptor-mediated excitation was increased in vCJD-inoculated mice. Using pharmacological experiments and computer simulations, we demonstrate that the increase in NMDA receptor-mediated excitation is necessary and sufficient to explain the distinctive bursting pattern in vCJD. These pathophysiological changes appear to result from a prion strain-specific gain-of-function and may explain some of the distinguishing clinical features of vCJD.
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Síndrome de Creutzfeldt-Jakob/fisiopatología , Modelos Animales de Enfermedad , Hipocampo/fisiología , Priones/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Potenciales Sinápticos/fisiología , Animales , Simulación por Computador , Síndrome de Creutzfeldt-Jakob/patología , Hipocampo/metabolismo , Hipocampo/patología , Humanos , Ratones , Ratones Noqueados , Ratones Transgénicos , Proteínas Priónicas , Priones/genética , Potenciales Sinápticos/genéticaRESUMEN
The membrane conductance of a pyramidal neuron in vivo is substantially increased by background synaptic input. Increased membrane conductance, or shunting, does not simply reduce neuronal excitability. Recordings from hippocampal pyramidal neurons using dynamic clamp revealed that adaptation caused complete cessation of spiking in the high conductance state, whereas repetitive spiking could persist despite adaptation in the low conductance state. This behavior was reproduced in a phase plane model and was explained by a shunting-induced increase in voltage threshold. The increase in threshold allows greater activation of the M current (I(M)) at subthreshold potentials and reduces the minimum adaptation required to stabilize the system; in contrast, activation of the afterhyperpolarization current is unaffected by the increase in threshold and therefore remains unable to stop repetitive spiking. The nonlinear interaction between shunting and I(M) has other important consequences. First, timing of spikes elicited by brief stimuli is more precise when background spikes elicited by sustained input are prohibited, as occurs exclusively with I(M)-mediated adaptation in the high conductance state. Second, activation of I(M) at subthreshold potentials, which is increased in the high conductance state, hyperpolarizes average membrane potential away from voltage threshold, allowing only large, rapid fluctuations to reach threshold and elicit spikes. These results suggest that the shift from a low to high conductance state in a pyramidal neuron is accompanied by a switch from encoding time-averaged input with firing rate to encoding transient inputs with precisely timed spikes, in effect, switching the operational mode from integration to coincidence detection.