Produce Action Potentials Research Articles

Neurophysiological modification of CA1 pyramidal neurons in a transgenic mouse expressing a truncated form of disrupted‐in‐schizophrenia 1

A t(1;11) balanced chromosomal translocation transects the Disc1 gene in a large Scottish family and produces genome-wide linkage to schizophrenia and recurrent major depressive disorder. This study describes our in vitro investigations into neurophysiological function in hippocampal area CA1 of a transgenic mouse (DISC1tr) that expresses a truncated version of DISC1 designed to reproduce aspects of the genetic situation in the Scottish t(1;11) pedigree. We employed both patch-clamp and extracellular recording methods in vitro to compare intrinsic properties and synaptic function and plasticity between DISC1tr animals and wild-type littermates. Patch-clamp analysis of CA1 pyramidal neurons (CA1-PNs) revealed no genotype dependence in multiple subthreshold parameters, including resting potential, input resistance, hyperpolarization-activated ‘sag’ and resonance properties. Suprathreshold stimuli revealed no alteration to action potential (AP) waveform, although the initial rate of AP production was higher in DISC1tr mice. No difference was observed in afterhyperpolarizing potentials following trains of 5–25 APs at 50 Hz. Patch-clamp analysis of synaptic responses in the Schaffer collateral commissural (SC) pathway indicated no genotype-dependence of paired pulse facilitation, excitatory postsynaptic potential summation or AMPA/NMDA ratio. Extracellular recordings also revealed an absence of changes to SC synaptic responses and indicated input–output and short-term plasticity were also unaltered in the temporoammonic (TA) input. However, in DISC1tr mice theta burst-induced long-term potentiation was enhanced in the SC pathway but completely lost in the TA pathway. These data demonstrate that expressing a truncated form of DISC1 affects intrinsic properties of CA1-PNs and produces pathway-specific effects on long-term synaptic plasticity.

Carbon Dioxide and Fruit Odor Transduction in Drosophila Olfactory Neurons. What Controls their Dynamic Properties?

We measured frequency response functions between odorants and action potentials in two types of neurons in Drosophila antennal basiconic sensilla. CO2 was used to stimulate ab1C neurons, and the fruit odor ethyl butyrate was used to stimulate ab3A neurons. We also measured frequency response functions for light-induced action potential responses from transgenic flies expressing H134R-channelrhodopsin-2 (ChR2) in the ab1C and ab3A neurons. Frequency response functions for all stimulation methods were well-fitted by a band-pass filter function with two time constants that determined the lower and upper frequency limits of the response. Low frequency time constants were the same in each type of neuron, independent of stimulus method, but varied between neuron types. High frequency time constants were significantly slower with ethyl butyrate stimulation than light or CO2 stimulation. In spite of these quantitative differences, there were strong similarities in the form and frequency ranges of all responses. Since light-activated ChR2 depolarizes neurons directly, rather than through a chemoreceptor mechanism, these data suggest that low frequency dynamic properties of Drosophila olfactory sensilla are dominated by neuron-specific ionic processes during action potential production. In contrast, high frequency dynamics are limited by processes associated with earlier steps in odor transduction, and CO2 is detected more rapidly than fruit odor.

Action Potential Energetics at the Organismal Level Reveal a Trade-Off in Efficiency at High Firing Rates

The energetic costs of action potential (AP) production constrain the evolution of neural codes and brain networks. Cellular-level estimates of AP-related costs are typically based on voltage-dependent Na(+) currents that drive active transport by the Na(+)/K(+) ATPase to maintain the Na(+) and K(+) ion concentration gradients necessary for AP production. However, these estimates of AP cost have not been verified at the organismal level. Electric signaling in the weakly electric fish Eigenmannia virescens requires that specialized cells in an electric organ generate APs with large Na(+) currents at high rates (200-600 Hz). We measured these currents using a voltage-clamp protocol and then estimated the energetic cost at the cellular level using standard methods. We then used this energy-intensive signaling behavior to measure changes in whole-animal energetics for small changes in electric discharge rate. At low rates, the whole-animal measure of AP cost was similar to our cellular-level estimates. However, AP cost increased nonlinearly with increasing firing rates. We show, with a biophysical model, that this nonlinearity can arise from the increasing cost of maintaining AP amplitude at high rates. Our results confirm that estimates of energetic costs based on Na(+) influx are appropriate for low baseline firing rates, but that extrapolating to high firing rates may underestimate true costs in cases in which AP amplitude does not decrease. Moreover, the trade-off between energetic cost and firing rate suggests an additional constraint on the evolution of high-frequency signaling in neuronal systems.

Expression of the voltage-gated potassium channel subunit Kv1.1 in embryonic zebrafish Mauthner cells

The escape response in zebrafish is mediated in part by the Mauthner cell and its two homologues, MiD2cm and MiD3cm. In adult fish, the Mauthner cell fires a single action potential when activated, while the homologs fire multiple action potentials. Voltage gated potassium channels containing the Kv1.1 subunit have been reported to play roles in modulating the firing properties of neurons. In this study we used a combination of techniques to determine if the Mauthner cells in embryonic zebrafish express Kv1.1. Our results using immunohistochemical and in situ hybridization experiments confirmed the expression of Kv1.1 in zebrafish reticulospinal neurons including the Mauthner cell. Current clamp recordings from the Mauthner cell showed that pharmacological block of Kv1.1 by the specific blocker, Dendrotoxin-K (DTXK), changed its firing properties from the production of a single action potential to firing multiple times. Together, these results suggest that Mauthner cells express potassium channels that contain Kv1.1 subunits, which might contribute to cell firing.

Stochastic Optimal Control as a Theory of Brain-Machine Interface Operation

The closed-loop operation of brain-machine interfaces (BMI) provides a framework to study the mechanisms behind neural control through a restricted output channel, with emerging clinical applications to stroke, degenerative disease, and trauma. Despite significant empirically driven improvements in closed-loop BMI systems, a fundamental, experimentally validated theory of closed-loop BMI operation is lacking. Here we propose a compact model based on stochastic optimal control to describe the brain in skillfully operating canonical decoding algorithms. The model produces goal-directed BMI movements with sensory feedback and intrinsically noisy neural output signals. Various experimentally validated phenomena emerge naturally from this model, including performance deterioration with bin width, compensation of biased decoders, and shifts in tuning curves between arm control and BMI control. Analysis of the model provides insight into possible mechanisms underlying these behaviors, with testable predictions. Spike binning may erode performance in part from intrinsic control-dependent constraints, regardless of decoding accuracy. In compensating decoder bias, the brain may incur an energetic cost associated with action potential production. Tuning curve shifts, seen after the mastery of a BMI-based skill, may reflect the brain's implementation of a new closed-loop control policy. The direction and magnitude of tuning curve shifts may be altered by decoder structure, ensemble size, and the costs of closed-loop control. Looking forward, the model provides a framework for the design and simulated testing of an emerging class of BMI algorithms that seek to directly exploit the presence of a human in the loop.

Voltage-gated sodium channel expression and action potential generation in differentiated NG108-15 cells

BackgroundThe generation of action potential is required for stimulus-evoked neurotransmitter release in most neurons. Although various voltage-gated ion channels are involved in action potential production, the initiation of the action potential is mainly mediated by voltage-gated Na+ channels. In the present study, differentiation-induced changes of mRNA and protein expression of Na+ channels, Na+ currents, and cell membrane excitability were investigated in NG108-15 cells.ResultsWhole-cell patch-clamp results showed that differentiation (9 days) didn’t change cell membrane excitability, compared to undifferentiated state. But differentiation (21 days) induced the action potential generation in 45.5% of NG108-15 cells (25/55 cells). In 9-day-differentiated cells, Na+ currents were mildly increased, which was also found in 21-day differentiated cells without action potential. In 21-day differentiated cells with action potential, Na+ currents were significantly enhanced. Western blot data showed that the expression of Na+ channels was increased with differentiated-time dependent manner. Single-cell real-time PCR data demonstrated that the expression of Na+ channel mRNA was increased by 21 days of differentiation in NG108-15 cells. More importantly, the mRNA level of Na+ channels in cells with action potential was higher than that in cells without action potential.ConclusionDifferentiation induces expression of voltage-gated Na+ channels and action potential generation in NG108-15 cells. A high level of the Na+ channel density is required for differentiation-triggered action potential generation.

The spike generator in the labellar taste receptors of the blowfly is differently affected by 4-aminopyridine and 5-hydroxytryptamine

In taste chemoreception of invertebrates the interaction of taste stimuli with specific membrane receptors and/or ion channels located in the apical membrane of taste receptor cells results in the generation of a receptor potential which, in turn, activates the ‘encoder’ region to produce action potentials which propagate to the CNS.This study investigates, in the labellar chemosensilla of the blowfly, Protophormia terraenovae, the voltage-gated K+ currents involved in the action potential repolarization and repetitive firing of the neurons by way of the Kv channel inhibitors, 4-aminopyridine and 5-hydroxytryptamine.The receptor potential and the spike activity were simultaneously recorded from the ‘salt’, ‘sugar’ and ‘deterrent’ cells, by means of the extracellular side-wall technique, in response to 150mM NaCl, 100mM sucrose and 1mM quinine HCl, before, 0÷10min after apical administration of 4-AP (0.01–10mM) or 5-HT (0.1–100mM).The results show that the receptor potential in all three cells is neither affected by 4-AP nor by 5-HT. Instead, spike activity is significantly decreased, by way of blocking different Kv channel types: an inactivating A-type K+ current (KA) modulating repetitive firing of the cells and responsible for the after hyperpolarization, and a sustained K+ current that resembles the delayed rectifier (DKR) and contributes to action potential repolarization.

Upper threshold of extracellular neural stimulation

It is well known that spiking neurons can produce action potentials in response to extracellular stimulation above certain threshold. It is widely assumed that there is no upper limit to somatic stimulation, except for cellular or electrode damage. Here we demonstrate that there is an upper stimulation threshold, above which no action potential can be elicited, and it is below the threshold of cellular damage. Existence of this upper stimulation threshold was confirmed in retinal ganglion cells (RGCs) at pulse durations ranging from 5 to 500 μs. The ratio of the upper to lower stimulation thresholds varied typically from 1.7 to 7.6, depending on pulse duration. Computational modeling of extracellular RGC stimulation explained the upper limit by sodium current reversal on the depolarized side of the cell membrane. This was further confirmed by experiments in the medium with a low concentration of sodium. The limited width of the stimulation window may have important implications in design of the electro-neural interfaces, including neural prosthetics.

Optical control after transfection of channelrhodopsin-2 recombinant adenovirus in visual cortical cells.

Channelrhodopsin-2 ectopically expressed in the retina can recover the response to blue light in genetically blind mice and rats, but is unable to restore visual function due to optic nerve or optic tract lesions. Long Evans rats at postnatal day 1 were used for primary culture of visual cortical cells, and 24 hours later, cells were transfected with recombinant adenovirus carrying channelrhodopsin-2 and green fluorescent protein genes. After 2–4 days of transfection, green fluorescence was visible in the cultured cells. Cells were stimulated with blue light (470 nm), and light-induced action potentials were recorded in patch-clamp experiments. Our findings indicate that channelrhodopsin-2-recombinant adenovirus transfection of primary cultured visual cortical cells can control the production of action potentials via blue light stimulation.

Membrane properties and electrogenesis in the distal axons of small dorsal root ganglion neurons in vitro

Although it is generally thought that sensory transduction occurs at or close to peripheral nerve endings, with action potentials subsequently propagating along the axons of dorsal root ganglia (DRG) neurons toward the central nervous system, the small diameter of nociceptive axons and their endings have made it difficult to estimate their membrane properties and electrogenic characteristics. Even the resting potentials of nociceptive axons are unknown. In this study, we developed the capability to record directly with patch-clamp electrodes from the small-diameter distal axons of DRG neurons in vitro. We showed using current-clamp recordings that 1) these sensory axons have a resting potential of -60.2 ± 1 mV; 2) both tetrodotoxin (TTX)-sensitive (TTX-S) and TTX-resistant (TTX-R) Na(+) channels are present and available for activation at resting potential, at densities that can support action potential electrogenesis in these axons; 3) TTX-sensitive channels contribute to the amplification of small depolarizations that are subthreshold with respect to the action potential in these axons; 4) TTX-R channels can support the production of action potentials in these axons; and 5) these TTX-R channels can produce repetitive firing, even at depolarized membrane potentials where TTX-S channels are inactivated. Finally, using voltage-clamp recordings with an action potential as the command, we confirmed the presence of both TTX-S and TTX-R channels, which are activated sequentially during action potential in these axons. These results provide direct evidence for the presence of TTX-S and TTX-R Na(+) channels that are functionally available at resting potential and contribute to electrogenesis in small-diameter afferent axons.

Integration of Subthreshold and Suprathreshold Excitatory Barrages along the Somatodendritic Axis of Pyramidal Neurons

Neurons integrate inputs arriving in different cellular compartments to produce action potentials that are transmitted to other neurons. Because of the voltage- and time-dependent conductances in the dendrites and soma, summation of synaptic inputs is complex. To examine summation of membrane potentials and firing rates, we performed whole-cell recordings from layer 5 cortical pyramidal neurons in acute slices of the rat's somatosensory cortex. We delivered subthreshold and suprathreshold stimuli at the soma and several sites on the apical dendrite, and injected inputs that mimic synaptic barrages at individual or distributed sites. We found that summation of subthreshold potentials differed from that of firing rates. Subthreshold summation was linear when barrages were small but became supralinear as barrages increased. When neurons were discharging repetitively the rules were more diverse. At the soma and proximal apical dendrite summation of the evoked firing rates was predominantly sublinear whereas in the distal dendrite summation ranged from supralinear to sublinear. In addition, the integration of inputs delivered at a single location differed from that of distributed inputs only for suprathreshold responses. These results indicate that convergent inputs onto the apical dendrite and soma do not simply summate linearly, as suggested previously, and that distinct presynaptic afferents that target specific sites on the dendritic tree may perform unique sets of computations.

EFFECT OF RANDOM LINK MALFUNCTIONS TO SYNCHRONIZATION OF HODGKIN–HUXLEY NEURONS IN A LATTICE NETWORK

We study the synchronization of Hodgkin–Huxley (HH) neurons connected in a directed lattice network under the influence of varying coupling strength (g) and random link malfunctions in the form of probabilistic deletion (probability q) and link direction flipping (p). We quantify the extent of synchronization of the neurons in the network by averaging the fraction of firing neurons that produce action potentials (exceeding a threshold potential of V th ) across the entire observation time. By extensively scanning over the values of g, synchronization of this network type can be enhanced by increasing g until it reaches a threshold value wherein synchronization will deteriorate abruptly due to suppression of neural firings. We also extensively probe the interplay of p and q and show that there are certain combinations for which the synchronization will improve, defying negative notions of how we perceive random malfunctions.

An impairment of cortical GABAergic neurons is involved in alkalosis-induced brain dysfunctions

Acid–base imbalance leads to pathological cognition and behaviors in the clinical practices. In the comparison with acidosis, the cellular mechanisms underlying alkalosis-induced brain dysfunction remain unclear. By using electrophysiological approach, we investigated the influences of high extracellular pH environment on cortical GABAergic neurons in terms of their responsiveness to synaptic inputs and their ability to produce action potentials. Artificial cerebral spinal fluid in high pH impairs excitatory synaptic transmission and spike initiation in cortical GABAergic neurons. The alkalosis-induced dysfunction of GABAergic neurons is associated with the decrease of receptor responsiveness and the increases of spike refractory periods and threshold potentials. Our studies reveal that alkalosis impairs cortical GABAergic neurons and subsequently deteriorate brain functions. The molecular targets for alkalosis action include glutamate receptor-channels and voltage-gated sodium channels on GABAergic neurons.

Acid Solution Is a Suitable Medium for Introducing QX-314 into Nociceptors through TRPV1 Channels to Produce Sensory-Specific Analgesic Effects

BackgroundPrevious studies have demonstrated that QX-314, an intracellular sodium channel blocker, can enter into nociceptors through capsaicin-activated TRPV1 or permeation of the membrane by chemical enhancers to produce a sensory-selective blockade. However, the obvious side effects of these combinations limit the application of QX-314. A new strategy for targeting delivery of QX-314 into nociceptors needs further investigation. The aim of this study is to test whether acidic QX-314, when dissolves in acidic solution directly, can enter into nociceptors through acid-activated TRPV1 and block sodium channels from the intracellular side to produce a sensory-specific analgesic effect.Methodology/Principal FindingsAcidic solution or noradrenaline was injected intraplantarly to induce acute pain behavior in mice. A chronic constrictive injury model was performed to induce chronic neuropathic pain. A sciatic nerve blockade model was used to evaluate the sensory-specific analgesic effects of acidic QX-314. Thermal and mechanical hyperalgesia were measured by using radiant heat and electronic von Frey filaments test. Spinal Fos protein expression was determined by immunohistochemistry. The expression of p-ERK was detected by western blot assay. Whole cell clamp recording was performed to measure action potentials and total sodium current in rats DRG neurons. We found that pH 5.0 PBS solution induced behavioral hyperalgesia accompanied with the increased expression of spinal Fos protein and p-ERK. Pretreatment with pH 5.0 QX-314, and not pH 7.4 QX-314, alleviated pain behavior, inhibited the increased spinal Fos protein and p-ERK expression induced by pH 5.0 PBS or norepinephrine, blocked sodium currents and abolished the production of action potentials evoked by current injection. The above effects were prevented by TRPV1 channel inhibitor SB366791, but not by ASIC channel inhibitor amiloride. Furthermore, acidic QX-314 employed adjacent to the sciatic nerve selectively blocked the sensory but not the motor functions in naïve and CCI mice.Conclusions/SignificanceAcid solution is a suitable medium for introducing QX-314 into nociceptors through TRPV1 channels to produce a sensory-specific analgesic effect.

Role of astrocytes in sensory processing in central nervous system

There are two types of cells in the central nervous systems (CNS) of mammals-neurons and glia. The structure and function of neurons have been thoroughly studied; while the role of glia in information processing has not been systematically studied because they cannot produce action potentials like neuron. During the past decades, glial cells were considered to play a supportive role in CNS instead of information processing. Recently, a variety of studies suggest that glial cells are actively involved in the regulation of brain function associated with neurons. Glial cells, especially astrocytes play important roles in different sensory processing. In the present article, we review the role of astrocytes in sensory processing in the CNS.

Novel Model to Investigate Blast Injury in the Central Nervous System

Blast-induced neurotrauma (BINT) is a common injury modality associated with the current war efforts and increasing levels of terrorist activity. Exposure to the primary blast wave generated by explosive devices causes significant neurological deficits and is responsible for many of the war-related pathologies. Despite research efforts, the mechanism of injury is still poorly understood. To this end, we have established a novel ex vivo model for the direct observation and quantification of BINT at the tissue level. The model provides a quantifiable and reproducible method to illustrate the mechanism of BINT. Isolated sections of guinea pig spinal cord white matter were exposed to a supersonic shockwave using a blast generator with small-scaled explosives. The blast wave impact with isolated tissue was observed using focused shadowgraphy with a high-speed camera recording at 90,000 fps. Concurrently, functional deficits were measured by monitoring the production of compound action potentials using a double sucrose gap-recording chamber. Additionally, anatomical deficits were measured after blast exposure with a dye exclusion assay to visualize axonal membrane permeability. Our findings demonstrate that direct exposure to the blast wave compressed nervous tissue at a rate of 60 m/sec and led to significant functional deficits. Damage to the isolated spinal cord was marked by increased axonal permeability, suggesting rapid compression from the shockwave-generated high strain rates that resulted in membrane disruption. The model provides new insight into the mechanism of BINT and permits direct observation that may contribute to the development of appropriate treatment regimens.

Channelopathies have many faces

A sodium channel known for its role in the perception of pain also seems to be necessary for olfaction. The multiple roles of this channel and the diverse effects of its mutations raise intriguing questions. See Article p.186 Humans and mice with mutations in the gene coding for the voltage-gated sodium ion channel Nav1.7, previously shown to be insensitive to pain, are now found to be unable to perceive odours. Olfactory sensory neurons that are missing this sodium channel still produce action potentials, but their synapses fail to transmit to downstream neuronal circuits. The Nav1.7-deficient phenotype of mice resembles that of human patients with Nav1.7 loss-of-function mutations, indicating that elimination of this ion channel creates a mouse model of congenital general anosmia.

Opioid Actions in Primary-Afferent Fibers—Involvement in Analgesia and Anesthesia

Opioids inhibit glutamatergic excitatory transmission from the periphery by activating G-protein coupled opioid receptors in the central terminals of primary-afferent neurons in the spinal substantia gelatinosa, resulting in antinociception. Opioid receptor activation in the peripheral terminals of primary-afferent neurons inhibits the production of action potentials in response to nociceptive stimuli given to the periphery, leading to antinociception. Opioids also exhibit a local anesthetic effect without opioid receptor activation in peripheral nerve fibers. This review article will focus on analgesia and anesthesia produced by the actions of opioids on primary-afferent fibers.

The dynamics of somatic input processing in spinal motoneurons in vivo

Uncovering how motoneurons utilize their voltage-sensitive conductances to systematically respond to a variety of inputs is paramount to understanding synaptic integration. In this study, we examine the input dynamics and frequency-dependent characteristics of active conductances in motoneurons as viewed from the soma in the decerebrate cat. We evaluated the somatic response of the motoneuron by superimposing a voltage sinus sweep (a sine wave in which frequency increases with time, which is often referred to as a zap or chirp) at a subset of membrane holding potentials during discontinuous, single-electrode, somatic voltage-clamp. Results from both experimental and modeling data indicate that ionic conductances can respond to a wide variety of input dynamics. Notably, it appears that there is a divergence between low input conductance type S and high input conductance type FF motoneurons in their response to input frequency. Type S motoneurons generate a larger response to lower frequency input dynamics (compared with their response to higher frequencies), whereas type FF generate a larger response to higher input frequency dynamics. Functionally, these results may indicate that motoneurons on the lower end of the motor pool (i.e., recruited first) may favor steady inputs, whereas motoneurons at the higher end (i.e., recruited later) may favor input transients in producing action potentials.

Frequency-dependent interaction of ultrashort E-fields with nociceptor membranes and proteins

We examined the influence of ultrashort pulses (USP) on sensory neurons. Single and high frequency bursts of 12 ns E-fields were presented to rat skin nociceptors that expressed distinct combinations of voltage-sensitive proteins. A single E-field pulse produced action potentials in all nociceptor subtypes at a critical threshold (E(c) ) of 403 V/cm. When configured into high frequency bursts, USP charge integrated to reduce the action potential threshold in a frequency and burst duration-dependent manner with E(c) as low as 16 V/cm (4000 Hz, 25 ms burst). There was no evidence of electroporation at field intensities near the E(c) for nociceptor activation. USP bursts activated a late, persistent Ca(++) flux that was identified as a dantrolene-sensitive Ca(++) -induced Ca(++) release (CICR). Influx of Ca(++) into the cell was required for the CICR and resulted in a reduction of the single pulse E(c) by about 50%.