The effect of plastic restructuring of the local chemoexcitable zone in the somatic membrane of the PPa4 neuron of Helix lucorum on the excitability of neighboring chemo- and electrically excited zones

Rhythmic application of acetylcholine or serotonin to the local zone of somatic membrane was used to study the effect of extinction of RPa4 neuron depolarization in Helix lucorum on the excitability of adjacent chemo- and electroexcitable zone. It has been found that the extinction of response to iontophoretic application of acetylcholine to one somatic zone decreases the sensitivity of serotonin and cholinoreceptors in adjacent zones, as well as the excitability of electroexcitable membrane. The effect on the excitability of adjacent zones does not depend on the type of receptors activated rhythmically, as the extinction of RPa4 response to the repeated application of serotonin also reduces the sensitivity of adjacent cholinoreceptor zones. A cause of this effect may lie in modification of chemoreceptors and ionic channels, by intracellular regulatory systems that become activated by repeated stimulation.     

Physical model of electroexcitable membranes. II. Mechanism of excitation]

The proposed model of action potential generation in electroexcitable membranes is based on physical phenomenon of the reversible thermal dielectric breakdown induced by injection of the activating charge carriers in the membrane under irritation. The model has been shown to describe well the main features of nerve excitation.     

Electrophysiology of identified neurosecretory and non-neurosecretory cells in the cockroach pars intercerebralis

Two cell types can be distinguished with intracellular recording from the pars intercerebralis of the American cockroach (Periplaneta americana). The first type, which corresponds morphologically to the medial neurosecretory cell, always had spontaneously occurring, overshooting action potentials. These action potentials are probably endogenously produced. Tetrodotoxin experiments revealed that sodium is the dominant ion of the action potential. The action potentials are followed by a relatively long after-hyperpolarization. The input resistance of these cells ranged from 120 to 390 M omega. A mathematical model, based on cellular morphology and response to current pulses, revealed a membrane time constant of about 100 msec and an axonal:somatic conductance ratio of approximately 13. Area-specific membrane resistance was estimated at 33 k omega cm2. These cells also often had reversible and spontaneous inhibitory postsynaptic potentials. The second cell type, which is non-neurosecretory, never produced spontaneous action potentials and rarely had synaptic potentials. Action potentials could be evoked by current injection into the cell body or by extracellular stimulation of their axons in the posteroventral portion of the the protocerebrum. These action potentials also depend on sodium ions. Their input resistance ranged from 16 to 35 M omega. They had a membrane time constant of approximately 15 msec and an axonal:somatic conductance ratio of about 9. Their area specific membrane resistance was estimated at 14 k omega cm2.     

Regulation of the plastic properties of an electro-excitable neuronal membrane by serotonin

The action of serotonin on plastic properties of electroexcitable membrane was studied in Helix lucorum parietal ganglia on neurones of two types: habituating (HC) and nonhabituating (NHC) to rhythmic intracellular stimulation by impulses of depolarizing current. Serotonin produced an effect of facilitation on HC (increase of responses to stimulation against the background of depolarization and rise of input resistance of the cell). Besides, serotonin completely blocked the ability of these cells to habituate to rhythmic stimulation. The obtained data testify that such action of serotonin may be based on suppression by it of C-dependent K-conductivity. Serotonin suppresses responses of NHC to stimulation and contributes to their habituation to rhythmic stimulation. Such action is due to serotonin suppression of Ca-conductivity, and, consequently, to elimination of the mechanism of action potential generation.     

Serotonin-induced changes in the responses of the electroexcitable membrane of snail neurons

The effects of serotonin (5-HT) added to the washing solution on the plastic properties of the electroexcitable membrane of nonidentified neurons of the parietal ganglion and identified neurons RPa2 are studied on the isolated nervous system of the snail. The neurons of the first group, which became rapidly habituated to the intracellular stimulation, are shown to manifest the ability to restore action potential generation in the presence of 5-HT. In contrast, the neurons possessing endogenous rhythmic activity (RPa2) in the presence of 5-HT generate burst activity against the background of the development of slow waves of the membrane potential. A comparative analysis of the effect of 5-HT and compounds with a known effect on Ca²⁺ and the calcium-dependent potassium channels (quinine, CoCl₂ and CdCl₂) points to the existence of various mechanisms providing for the serotoninergic regulation of the plasticity of the electroexcitable neuron membrane. This difference stems from the processes of activation and blockade of the Ca-dependent mechanisms.A. I. Karaev Institute of Physiology Azerbaizhan Academy of Sciences, Baku. Translated from Neirofiziologiya, Vol. 24, No. 3, pp. 286–290, May–June, 1992.     

Evidence that glycine and GABA mediate postsynaptic inhibition of bulbar respiratory neurons in the cat.

Experiments were carried out on decerebrate cats to identify transsynaptic mediators of spontaneous postsynaptic inhibition of bulbar inspiratory and postinspiratory neurons. Somatic membrane potentials were recorded through the central micropipette of a coaxial multibarreled electrode. Blockers of type A gamma-aminobutyric acid (GABA-A) and glycine receptors were iontophoresed extracellularly from peripheral micropipettes surrounding the central pipette. Effective antagonism was demonstrated by iontophoresis of agonists with antagonists; application of strychnine antagonized the action of glycine but not GABA, and application of bicuculline antagonized the action of GABA but not glycine. In both types of neurons, iontophoresis of either antagonist depolarized the somatic membrane and increased input resistance throughout the respiratory cycle. Bicuculline preferentially depolarized the somatic membrane in both types of neurons during inactive phases. Strychnine increased the firing rate of inspiratory neurons during inspiration despite maintenance of somatic membrane potential at preiontophoresis levels. Tetrodotoxin reduced the effects of iontophoresed bicuculline and strychnine, suggesting that the action of the antagonists required presynaptic axonal conduction. The present results suggest that presynaptic release of both GABA and glycine contributes to tonic postsynaptic inhibition of bulbar respiratory neurons. GABA-A receptors appear to contribute to inhibition during inactive phases in inspiratory and postinspiratory neurons, whereas glycinergic mechanisms appear to contribute to inspiratory inhibition in inspiratory neurons.     

Regulation of the neuronal functional state by ultra low doses of different biologically active substances. Nonspecific effect

The role of biologically active substances in ultra-low doses (10(-15)-10(-27) mol/l) is discussed from the different points of view. The most detailed analysis of neurobiological effects produced by these doses can be studied on the preparate of completely isolated molluscan neurones. In this case the possibility arises to control the first modifications of action at the electrophysiological characteristics of neuronal activity. These changes of electrical activity can be regarded as a reaction to biologically active substance. The following characteristics were controlled: the level of membrane resting potential (MP), the electroexcitable membrane and pacemaker mechanism, chemical sensitivity of somatic membrane loci to neurotransmitter acetylcholine (Ach). Several substances were used in these experiments: two kinds of synthetic antioxidant, GABA, ethanol, serotonine, DSIP (delta-sleep inducing peptide), antibiotic ruboxil, nootrop GVS-111. The isolated neurones were placed into the special chamber. All these substances (0.35 ml) were added single dosing into this chamber with living physiological solution in concentration 10(-15)-10(-27) mol/l. The results demonstrated that all substances had initiated the development of prolonged neurophysiological responses. The intensities of neuronal reactions didn't depend in contact period on the concentration and on the type of substance. It is suggested that these data reveal the existence of unknown modes of regulation of neuronal functional states and presence of hidden channel for information transfer and receiving. This different way of regulation is extremely important influence living organisms.     

Ca2+-activated K+ currents regulate odor adaptation by modulating spike encoding of olfactory receptor cells

The olfactory system is thought to accomplish odor adaptation through the ciliary transduction machinery in olfactory receptor cells (ORCs). However, ORCs that have lost their cilia can exhibit spike frequency accommodation in which the action potential frequency decreases with time despite a steady depolarizing stimulus. This raises the possibility that somatic ionic channels in ORCs might serve for odor adaptation at the level of spike encoding, because spiking responses in ORCs encode the odor information. Here I investigate the adaptational mechanism at the somatic membrane using conventional and dynamic patch-clamp recording techniques, which enable the ciliary mechanism to be bypassed. A conditioning stimulus with an odorant-induced current markedly shifted the response range of action potentials induced by the same test stimulus to higher concentrations of the odorant, indicating odor adaptation. This effect was inhibited by charybdotoxin and iberiotoxin, Ca2+-activated K+ channel blockers, suggesting that somatic Ca2+-activated K+ currents regulate odor adaptation by modulating spike encoding. I conclude that not only the ciliary machinery but also the somatic membrane currents are crucial to odor adaptation.     

Nerve cells of Helix pomatia during hypercapnia

Hypercapnia leading initially to activation of synaptic activity and of pace-maker mechanism is accompanied later by inactivation of the electro-excitable membrane and is completed by loss of chemosensitivity. Restoration of the properties of the neurone following transition to normal atmospheric air develops in reverse order. All hypercapnic changes of the properties of the electroexcitable and chemosensitive membrane take place at a constant level of the membrane potential which points to stability of K-Na-pump.     

Multiplicity of pacemaker zones in Helix pomatia neurons

Intracellular microelectrode recordings from neurons ofHelix pomatia revealed several local zones of action potential generation both on the soma and on some of the branches of the neurons. Under certain conditions the activity of individual loci of the neuron membrane was synchronized to produce a normal action potential. It is suggested that the somatic membrane of neurons is heterogeneous in structure and consists of separate loci of an electrically excitable membrane, incorporating active and latent pacemaker zones. Neurons ofH. pomatia are characterized by two types of action potential with different triggering mechanisms: one (synaptic) type is generated under the influence of the EPSP, the other (pacemaker) arises through activation of endogenous factors for the neuron (pacemaker potentials). The interaction between synaptic and pacemaker potentials during integrative activity of the neuron is discussed.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 5, No. 1, pp. 88–94, January–February, 1973.     

Effect of FMRFamide on the electrical and plastic properties of identified Helix pomatia neurons

Effect of the neuropeptide FMRFamide on two types of nerve cells differing in plastic properties: habituating and non-habituating to rhythmic intracellular stimulation, has been studied. FMRFamide causes a slow developing, continuously growing depolarisation and an increase of input resistance of the most part of habituating cells resulting in inhibition of their habituation to intracellular stimulation. No desensitisation of cells to the action of FMRFamide was observed. The data obtained by using Ca-ionophore, imidazole and caffeine show that the effect of the peptide may be caused by inhibition of Ca-dependent K-conductance and depends on cAMP metabolism. FMRFamide exerts a less pronounced action on non-habituating cells and does not change their plastic properties. Inhibition by FMRFamide of the habituation at the level of electroexcitable membrane may play a significant part in regulation of neuronal plasticity.     

Dopamine modification of the Ca-component of the action potential in the neuron soma of the mollusk L. stagnalis

Dopamine has been found to decrease reversibly the action potential rising phase in identified L. stagnalis neurones. Using voltage-clamp and intracellular dialysis techniques it is shown that dopamine inhibits electroexcitable Ca-current, and changes in Ca-current determine those in the action potential shape. Serotonin and adrenaline influence the Ca-current in a similar way as dopamine does. Inhibition of Ca-conductance is found not to be related to possible effect of these neuromediators on intracellular level of free Ca ions or cyclic AMP.     

A Time Course Analysis of the Electrophysiological Properties of Neurons Differentiated from Human Induced Pluripotent Stem Cells (iPSCs)

Many protocols have been designed to differentiate human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) into neurons. Despite the relevance of electrophysiological properties for proper neuronal function, little is known about the evolution over time of important neuronal electrophysiological parameters in iPSC-derived neurons. Yet, understanding the development of basic electrophysiological characteristics of iPSC-derived neurons is critical for evaluating their usefulness in basic and translational research. Therefore, we analyzed the basic electrophysiological parameters of forebrain neurons differentiated from human iPSCs, from day 31 to day 55 after the initiation of neuronal differentiation. We assayed the developmental progression of various properties, including resting membrane potential, action potential, sodium and potassium channel currents, somatic calcium transients and synaptic activity. During the maturation of iPSC-derived neurons, the resting membrane potential became more negative, the expression of voltage-gated sodium channels increased, the membrane became capable of generating action potentials following adequate depolarization and, at day 48–55, 50% of the cells were capable of firing action potentials in response to a prolonged depolarizing current step, of which 30% produced multiple action potentials. The percentage of cells exhibiting miniature excitatory post-synaptic currents increased over time with a significant increase in their frequency and amplitude. These changes were associated with an increase of Ca²⁺ transient frequency. Co-culturing iPSC-derived neurons with mouse glial cells enhanced the development of electrophysiological parameters as compared to pure iPSC-derived neuronal cultures. This study demonstrates the importance of properly evaluating the electrophysiological status of the newly generated neurons when using stem cell technology, as electrophysiological properties of iPSC-derived neurons mature over time.     

The induction of periodic and chaotic activity in a molluscan neurone

During prolonged exposure to extracellular 4-aminopyridine (4 AP) the periodic activity of the somatic membrane of an identified molluscan neurone passes from a repetitive regular discharge of >90 mV amplitude action potentials, through double discharges to <50 mV amplitude oscillations. Return to standard saline causes the growth of parabolic amplitude-modulated oscillations that develop, through chaotic amplitude-modulated oscillations, into regular oscillations. These effects are interpreted in terms of the actions of 4 AP on the dynamics of the membrane excitation equations.Emma and Leslie Reid scholar     

Propagation of action potentials in squid giant axons. Repetitive firing at regions of membrane inhomogeneities

Effects of reduction in potassium conductance on impulse conduction were studied in squid giant axons. Internal perfusion of axons with tetraethylammonium (TEA) ions reduces G K and causes the duration of action potential to be increased up to 300 ms. This prolongation of action potentials does not change their conduction velocity. The shape of these propagating action potentials is similar to membrane action potentials in TEA. Axons with regions of differing membrane potassium conductances are obtained by perfusing the axon trunk and one of its two main branches with TEA after the second branch has been filled with normal perfusing solution. Although the latter is initially free of TEA, this ion diffuses in slowly. Up until a large amount of TEA has diffused into the second branch, action potentials in the two branches have very different durations. During this period, membrane regions with prolonged action potentials are a source of depolarizing current for the other, and repetitive activity may be initiated at transitional regions. After a single stimulus in either axon region, interactions between action potentials of different durations usually led to rebound, or a short burst, of action potentials. Complex interactions between two axon regions whose action potentials have different durations resembles electric activity recorded during some cardiac arrhythmias.     

神经元放电阈值的可变性及其意义

神经元能够将不同时空模式的突触输入转化为时序精确的动作电位输出，这种灵活、可靠的信息编码方式是神经集群在动态环境或特定任务下产生所需活动模式的重要基础。动作电位的产生遵循全或无规律，只有当细胞膜电压达到放电阈值时，神经元才产生动作电位。放电阈值在细胞内和细胞间具有高度可变性，具体动态依赖于刺激输入和放电历史。特别是，放电阈值对动作电位起始前的膜电压变化十分敏感，这种状态依赖性产生的生物物理根源包括Na⁺失活和K⁺激活。在绝大多数神经元中，动作电位的触发位置是轴突起始端，这个位置处的阈值可变性是决定神经元对时空输入转化规律的关键因素。但是，电生理实验中动作电位的记录位置却通常是胞体或近端树突，此处的阈值可变性高于轴突起始端，而其产生的重要根源是轴突动作电位的反向传播。基于胞体测量的相关研究显示，放电阈值动态能够增强神经元的时间编码、特征选择、增益调控和同时侦测能力。本文首先介绍放电阈值的概念及量化方法，然后详细梳理近年来国内外关于放电阈值可变性及产生根源的研究进展，在此基础上归纳总结放电阈值可变性对神经元编码的重要性，最后对未来放电阈值的研究方向进行展望。     

Activation of Nuclear Calcium Dynamics by Synaptic Stimulation in Cultured Cortical Neurons

L-type voltage-sensitive Ca2+ channels (VSCCs) are enriched on the neuronal soma and trigger gene expression during synaptic activity. To understand better how these channels regulate somatic and nuclear Ca2+ dynamics, we have investigated Ca2+ influx through L-type VSCCs following synaptic stimulation, using the long-wavelength Ca2+ indicator fluo-3 combined with laser scanning confocal microscopy. Single synaptic stimuli resulted in rapid Ca2+ transients in somatic cytoplasmic compartments (<5 ms rise time). Nuclear Ca2+ elevations lagged behind cytoplasmic levels by approximately 60 ms, consistent with a dependence on diffusion from a cytoplasmic source. Pharmacological experiments indicated that L-type VSCCs mediated approximately 50% of the nuclear and somatic (cytoplasmic) Ca2+ elevation in response to strong synaptic stimulation. In contrast, relatively weak excitatory postsynaptic potentials (EPSPs; approximately 15 mV) or single action potentials were much less effective at activating L-type VSCCs. Antagonist experiments indicated that activation of the NMDA-type glutamate receptor leads to a long-lasting somatic depolarization necessary to activate L-type VSCCs effectively during synaptic stimuli. Simulation of action potential and somatic EPSP depolarization using voltage-clamp pulses indicated that nuclear Ca2+ transients mediated by L-type VSCCs were produced by sustained depolarization positive to -25 mV. In the absence of synaptic stimulation, action potential stimulation alone led to elevations in nuclear Ca2+ mediated by predominantly non-L-type VSCCs. Our results suggest that action potentials, in combination with long-lived synaptic depolarizations, facilitate the activation of L-type VSCCs. This activity elevates somatic Ca2+ levels that spread to the nucleus.     

Initiation of Sodium Spikelets in Basal Dendrites of Neocortical Pyramidal Neurons

Cortical information processing relies critically on the processing of electrical signals in pyramidal neurons. Electrical transients mainly arise when excitatory synaptic inputs impinge upon distal dendritic regions. To study the dendritic aspect of synaptic integration one must record electrical signals in distal dendrites. Since thin dendritic branches, such as oblique and basal dendrites, do not support routine glass electrode measurements, we turned our effort towards voltage-sensitive dye recordings. Using the optical imaging approach we found and reported previously that basal dendrites of neocortical pyramidal neurons show an elaborate repertoire of electrical signals, including backpropagating action potentials and glutamate-evoked plateau potentials. Here we report a novel form of electrical signal, qualitatively and quantitatively different from backpropagating action potentials and dendritic plateau potentials. Strong glutamatergic stimulation of an individual basal dendrite is capable of triggering a fast spike, which precedes the dendritic plateau potential. The amplitude of the fast initial spikelet was actually smaller that the amplitude of the backpropagating action potential in the same dendritic segment. Therefore, the fast initial spike was dubbed “spikelet”. Both the basal spikelet and plateau potential propagate decrementally towards the cell body, where they are reflected in the somatic whole-cell recordings. The low incidence of basal spikelets in the somatic intracellular recordings and the impact of basal spikelets on soma-axon action potential initiation are discussed.     

Dendritic action potentials of pyramidal tract neurons in the cat sensorimotor cortex

The intracellular activity of pyramidal tract neurons was studied during electrical stimulation of ventrolateral and ventroposterolateral thalamic nuclei in acute experiments on cats immobilized by myorelaxants. Somatic action potentials were observed and spontaneous spikes were also produced by single and rhythmic stimulation of the thalamic nuclei at the rate of 8–14 Hz, by iontophoretic application of strychnine, and by intracellular depolarizing current pulses. These potentials had a relatively low and variable amplitude of 5–60 mV and are presumed to be dendritic action potentials. It is postulated that these variable potentials arise in the dendrites of pyramidal neurons with multiple zones generating such activity. No interaction was observed where somatic and dendritic action potentials occur simultaneously. The possible functional role of dendritic action potentials is discussed.I. S. Beritashvili Institute of Physiology, Academy of Sciences of the Georgian SSR, Tbilisi. Translated from Neirofiziologiya, Vol. 18, No. 4, pp. 435–443, July–August, 1986.     

Recovering the passive properties of tapered dendrites from single and dual potential recordings

We demonstrate that measurement of the membrane potential at one or more sites on a branched and tapered neuron following a known transient injection of subthreshold somatic current uniquely determines the cell's passive electrical properties. That is, knowledge of the potentials allows recovery of the cell's axial resistance, membrane capacitance, membrane conductance and soma conductance. The argument underlying uniqueness leads immediately to a constructive, robust algorithm that we successfully test on synthetic data. The robustness stems from the fact that the algorithm requires only a few weighted integrals, or moments, of the measured potentials.