Neuronal response in a strychninized cortical slab isolated from the cat

Intracellular response in neurons and glial cells of an isolated cortical slab to direct electrical stimulation of the slab following surface application of strychnine was investigated during experiments on immobilized unanesthetized cats. Strychnine induced single epileptiform discharges and after-discharges in the slab and in the neurons it contained in the form of large-scale paroxysmal depolarization shifts (PDS) in membrane potential (MP). Spontaneous summated epileptiform discharges and neuronal activity in the units examined were not very synchronized. Electrical stimulation induced generalized paroxysmal activity in the isolated slab. Neuronal PDS were accompanied by refractory periods, onset of which did not depend on MP level. Strychnine increased the number of neurons manifesting background activity in which action potentials were generated by rhythmic depolarizing MP waves of extrasynaptic origin. Epileptiform response in strychninized cortical isolated slabs to presentation of single stimuli is accompanied by major depolarization shifts in the MP of glial cells. Paroxysmal excitation is thought to be triggered in strychninized isolated cortical slabs by extrasynaptic factors and closely linked to altered concentration of extracellular potassium.I. I. Mechnikov University, Odessa. Translated from Neirofiziologiya, Vol. 22, No. 1, pp. 23–29, January–February, 1990.     

Post-tetanic changes in intracellular response to direct cortical stimulation

Changes in responses to single stimulations of the cortical surface after tetanization (frequency 50/sec, duration 1–10 sec) were studied in sensorimotor cortical neurons of an unanesthetized rabbit on intracellular and "quasiintracellular" recordings. After tetanization insufficient to generate epileptiform after-discharges, an increase was observed in the amplitude and duration of exciting postsynaptic potentials (EPSP) induced by a single test stimulus. This increase is considered as post-tetanic potentiation (PTP). Its duration did not exceed 1 min. The amplitude of inhibitory postsynaptic potentials (IPSP) showed a considerably smaller increase or did not change or even decreased. The PTP increased with an increase in the strength and duration of the tetanization, reaching especially high values during tetanization sufficiently intensive to evoke epileptiform after-discharges. In this case the response to a single test stimulus was identical to an epileptiform intracellular discharge. The data obtained confirm the important role of PTP of the exciting synapses in the generation of epileptiform after-discharges. A simple model of a neuron network with exciting and inhibiting feedbacks which accounts for the generation of epileptiform activity is examined.Institute of the Brain, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 2, No. 6, pp. 601–610, November–December, 1970.     

Postsynaptic components of paroxysmal neuronal response in the strychninized neocortex

Neuronal response in the strychninized cortical suprasylvian gyrus was investigated in experiments on immobilized and unanesthetized cats using intracellular techniques. Paroxysmal depolarizing shifts (PDS) in neuronal membrane potential were recorded, consisting of a bursting discharge and slow depolarization wave. It was found when using intracortical stimulation that PDS can accumulate and change in shape and size. Bursting discharges in PDS were induced by large-scale EPSP which could be distinguished from paroxysmal response. Data from presumably intradendritic readings demonstrated the presence of large-scale EPSP during the generation of epileptiform discharges in the cortex. In a proportion of cells, PDS were accompanied by hyperpolarizing potentials — apparently IPSP, since they undergo reversal with intercellular administration of Cl^–. The contribution of excitatory and inhibitory synaptic influences to paroxysmal neuronal response is discussed.I. I. Mechnikov State University, Odessa. Translated from Neirofiologiya, Vol. 22, No. 5, pp. 642–649, September–October, 1990.     

Simulation of bursting activity in theHelix RPa1 neuron: Role of calcium ions

A model of bursting activity in the RPal neuron of the snailHelix pomatia has been developed. In this model, calcium conductances do not play a key role in generation of slow oscillations of membrane potential (MP). The possibility of simulating the maintenance of bursting in the presence of cadmium ions is shown. Inclusion in the model of the calcium-inactivated calcium conductance makes it possible to reproduce both adaptation of the neuron to constant polarizing current, which modifies bursting, and the development of slow inward current when MP is clamped at different phases at the slow wave. In our simulations, the characteristic properties of bursts (such as an increase in the frequency of action potentials and a decrease in spike undershoot at the beginning of a burst) are due to the cumulative inactivation of potassium current. The advantages of the presented mathematical model of bursting compared with other models are discussed.Neirofiziologiya/Neurophysiology, Vol. 26, No. 5, pp. 373–381, September–October, 1994.     

Protracted depolarizing potentials of neurons in a strychninized cortical slab isolated from the cat

Neuronal response in a cortical slab isolated from the cat during surface application of strychnine was investigated in experiments on immobilized unanesthetized animals by means of intracellular recording techniques. Protracted depolarizing potentials (PDP) were found to occur spontaneously and in response to a single intracortical electrical stimulus in a proportion of the neurons. These potentials could be triggered by transformation of response along the lines of "paroxysmal depolarizing shift" (PDS) — hyperpolarization, with hyperpolarization replaced by depolarizing potentials. A further increase in depolarizing after-potentials resulted in the generation of PDP. These changes were normally accompanied by enhanced summated epileptiform activity in the isolated cortical slab. It is postulated that PDP were triggered by increased calcium conductance at the neuronal membrane during intensification of paroxysmal response in the isolated cortical slab.I. I. Mechnikov University, Odessa. Translated from Neirofiziologiya, Vol. 22, No. 1, pp. 19–23, January–February, 1990.     

Effect of substances blocking calcium channels (verapamil,D-600, manganese ions) on transmitter release from motor nerve endings in frog muscle

Experiments on isolated frog nerve-muscle preparations showed that manganese ions (0.4–5.0 mM) inhibit evoked transmitter release by reducing the quantum composition of the end-plate potentials, and they intensify spontaneous transmitter release to a certain extent by increasing the frequency of miniature potentials. Verapamil (1 · 10^–6–5·10^–5 g/ml) and D-600 (2.5·10^–5 g/ml), by contrast with manganese ions, do not inhibit evoked release, but also intensify spontaneous release of the transmitter. All the agents tested prevent the potentiating effect of imidazole (3 mM). During repetitive stimulation, verapamil disturbs action potential generation in the motor nerve. Manganese ions had no such action. It is concluded that between the calcium channels of motor nerve endings and the calcium channels of heart muscle or the neuron soma there are molecular differences, expressed as sensitivity to the blocking action of verapamil and D-600.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 9, No. 4, pp. 415–422, July–August, 1977.     

A study of the time and frequency characteristics of the potentials evoked in the acoustical cortex

Evoked potentials in the auditory cortex of the cat are measured by applying auditory stimulations in the form of tone bursts of 700 Hz. Transient evoked potentials obtained in this way are transformed to the frequency domain using a Laplace Transform. The amplitude frequency characteristic obtained with this semi-empirical method depicts maxima of EEG-amplitude in frequency ranges of 10–13 Hz and 60–80 Hz. The correlation between the time course of evoked potentials and spontaneous activity of the brain and the efficiency of the method used are pointed out.     

Neuronal response in an epileptically excited isolated cortical slab to single electrical stimuli

Neuronal response to single stimuli was investigated in a cortical slab isolated from immobilized cats before, during, and after onset of induced epileptic states. Neurons of the isolated cortical slab were found to generate EPSP and paroxysmal depolarizing shifts (PDS) in membrane potential (MP) during the development of generalized epileptoid activity; these occurred together with refractory periods. Duration of the latter corresponds with the PDS plateau and repolarizing shifts in MP. Single electrical stimuli induced gradual alteration in PDS as these shifts developed. Neurons still maintain their ability to generate PDS arising in response to presentation of single stimuli once ictal activity has ceased. Postsynaptic response is not thought to play a decisive role in the genesis of epileptoid activity. Nonspecific factors and especially alterations in the concentration of electrogenic ions apparently contribute to this phenomenon.I. I. Mechnikov State University, Odessa. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 198–204, March–April, 1989.     

Effect of penicillin on synaptic activity of motoneurons in the lamprey isolated spinal cord

The effect of penicillin on the membrane potential (MP) and synaptic activity of motoneurons in the isolated spinal cord of the river lamprey was investigated. In cells with a low initial MP (58.7±5.2 mV, n=28), penicillin (2.5 mmole·liter^–1) caused a depolarization, and potentiated excitatory postsynaptic potentials (EPSPs) that were evoked by stimulating spinal tracts and dorsal roots. The EPSPs were potentiated by 80–220% relative to their initial amplitude. In motoneurons with a higher MP (72.0±5.7 mV, n=20), a depolarization did not develop, and the potentiation of EPSPs did not exceed 25–70%. The effects of penicillin were inhibited when antagonists of excitatory and inhibitory amino acids were added to the superfusate. The results obtained imply that the motoneuron membranes have two acceptor sites for penicillin.Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Saint Petersburg Institute of Biological Research, Belgrade, Yugoslavia. Translated from Neirofiziologiya, Vol. 24, No. 2, pp. 151–160, March–April, 1992.     

The ionic regulation of action potentials in the axon of a stretch receptor neuron of the stick insect (Carausius morosus)

Summary The ionic requirements for the action potentials recorded from the axon of the dorsal longitudinal stretch receptor inCarausius morosus have been studied using extracellular electrodes.In the intact preparation prolonged exposure to sodium-free, calcium-free, or magnesium-free salines produces no observable change in the amplitude of action potentials. Similarly, tetrodotoxin (1×10^–6 M) and cobaltous chloride (1×10^–2 M) are both ineffective in blocking the action potentials.In preparations in which the ionic barrier has been disrupted by removal of the nerve sheath the action potentials show sodium dependence. They are sustained in high sodium salines (150 mM) but are reversibly abolished in sodium-free salines. They are also reversibly abolished in 1×10^–6 M TTX, but unaffected by calcium-free or magnesium-free salines, or by cobaltous chloride (1×10^–2 M).It is concluded that the action currents in the axon of the stretch receptor are carried by sodium ions.     

Effect of exogenous acetylcholine on potassium currents in the frog motor nerve ending during acetylcholinesterase inhibition

The effect of exogenous acetylcholine (ACh) on potassium currents in the motor nerve ending (NE) has been studied in neuromuscular preparations of the frog cutaneous-sternal muscle by extracellular recording of evoked electrical potentials from the NE. The investigation was performed during inhibition of acetylcholinesterase (AChE) activity by specific inhibitors and AChE removal from the synaptic cleft by collagenase. After AChE inhibition by either armine or proserine, or after treatment of the preparation with collagenase, no effect of exogenous ACh in concentrations of 1·10^–4–6·^–4 mole/liter was observed, in contrast to results from preparations with intact AChE. However, under the same conditions, as in the case of active AChE, ACh in concentrations of 7·10^–4–2·10^–3 mole/liter inhibited Ca-activated potassium current of the NE membrane. Experiments with dipyroxim, a synaptic AChE reactivator, have shown that the ACh effect on the potential-dependent potassium current is mediated by specific AChE. The role of AChE is discussed in respect to its significance for realization of the ACh action on potential-dependent potassium current in NE.Translated from Neirofiziologiya, Vol. 25, No. 2, pp. 146–149, March–April, 1993.     

Dynamics of cortical unit responses during defensive conditioning to sound

Several phases were distinguished in single-unit responses in areas 3 and 4 during defensive conditioning to acoustic stimulation: an initial response, short inhibition of the spike discharge, early and late after-discharges, and changes arising after the end of acoustic stimulation. The initial spike response appeared or intensified (if present already) in the first period of defensive conditioning parallel with an increase in spontaneous unit activity. After-discharges appeared later. The conditioned-reflex movement usually began 100–400 msec after stimulation began. This latent period of the first movement was the same whether for a real conditioned reflex or an after-discharge. Comparison of the latent periods of conditioned movements with the phases of the unit responses showed that the conditioned responses of the cortical neuron were primarily modified after-discharges of neurons evoked by a conditioned stimulus. Differential unit responses to acoustic stimulation, also based on after-discharges, were formed just as actively as positive. The basic role of reinforcement during conditioning is not to increase the excitability of the neurons, which is important in connection with their acquisition of polysensory properties, but to modify the after-discharges of the neurons.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 10, No. 4, pp. 339–347, July–August, 1978.     

Structural and developmental differences between three types of Na channels in dorsal root ganglion cells of newborn rats

Summary The changes in Na current during development were studied in the dorsal root ganglion (DRG) cells using the whole-cell patch-clamp technique. Cells obtained from rats 1–3 and 5–8 days after birth were cultured and their Na currents were compared. On top of the two types of Na currents reported in these cells (fast-FA current and slow-S current) a new fast current was found (FN). The main characteristics of the three currents are: (i) The voltages of activation are –37, –36, and –23 mV for the FN, FA and S currents, respectively. (ii) The activation and inactivation kinetics of FN and FA currents are about five times faster than those of the S current. (iii) The voltages at which inactivation reaches 50% are –139, –75 and –23 mV for the FN, FA and S currents, respectively.The kinetics and voltage-dependent parameters of the three currents and their density do not change during the first eight days after birth. However, their relative frequency in the cells changes. In the 1–3 day-old rats the precent of cells with S, FA, and mixed S+FN currents is 22, 18, and 60% of the cells, respectively. In the 5–8 day-old, the percent of cells with S, FA, and FN+S is 10, 66 and 22%. The relative increase in the frequency of cells with FA current during development can contribute to the ease of action potential generation compared with cells with FN currents, which are almost completely inactivated under physiological conditions. The predominance of FA cells also results in a significant decrease in the relative frequency of cells with the high-threshold, slow current.Antibodies directed against a part of the S₄ region of internal repeat I of the sodium channel (C ₁ ⁺ , amino acids 210–223, eel channel numbering) were found to shift the voltage dependence of FA current inactivation (but not of FN or S currents) to more negative potentials. The effect was found only when the antibodies were applied externally. The results suggest that FN, FA and S types of Na currents are generated by channels, which are different in the topography of the C ₁ ⁺ region in the membrane.     

Modulation of electrical activity of neuron RPa2 ofHelix pomatia

Neuron RPa2 ofHelix pomatia can generate rhythmic (beating) or periodic (bursting) activity. A spontaneous switch from beating to bursting activity takes place in the course of tens of minutes. Similar changes in electrical activity can be induced by the addition of the water-soluble fraction obtained from a homogenate of snail ganglia to the experimental chamber. Artificial polarization of the membrane of neuron RPa2 by asteady inward current leads to an increase in the duration of intervals between bursts and to a decrease in the number of action potentials in the burst. With an increase in amplitude of the polarizing current, action potential generation ceases completely, but generation of waves of membrane potential persists. If the voltage on the neuron membrane is clamped, periodic fluctuations of membrane current disappear. It is suggested that action potential generation by neurons RPa2 is determined by the properties of the potential-dependent conductance of its membrane, i.e., that it is endogenous in origin and can be regulated by compounds acting on the membrane. These compounds, secreted by other neurons, resemble neurotransmitters or neurohormones.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 13, No. 4, pp. 406–412, July–August, 1981.     

The uptake and incorporation of leucine and cystine by the mycelial and yeastlike phases of Blastomyces dermatitidis

Uptake and incorporation of L-leucine-C¹⁴ and L-cystine-S³⁵ was studied in the mycelial [MP] and yeastlike [YP] phases of the dimorphic fungal pathogen,Blastomyces dermatitidis. Both amino acids entered the cells of the two morphological forms ofB. dermatitidis by a permease-like system at low external concentrations of substrate. At high substrate levels, the amino acids entered the cells by a simple diffusion-like process in addition to the permease-like system. Michaelis-Menten constants [K_m] for L-leucine was found to be 1.1×10^–5 M and 4.4×10^–5 M for the MP and YP phases, respectively. The K_m for L-cystine was found to be 1.0×10^–5 M for the MP and 0.5×10^–5 M for the YP. A requirement for energy supplied by metabolic activity was demonstrated by the inhibition of uptake and incorporation of the amino acids by cells incubated with either 2,4-dinitrophenol or sodium azide. Amino acid uptake was broadly tolerant of hydrogen ion concentration, but definite optima were demonstrated at pH 7.0 to 7.5.     

An hypothesis for the interpretation of the contractile response of vascular smooth muscle at the cellular level

The long preservation and recovery of functional (contractile) properties in cultured aortic smooth muscle cells, even after replating or deep-frozen storage and the measurement of their responses are now technically settled issues. We could thus study extensively the responses of single cultured cells from rat thoracic aorta. Responses were elicited by the addition of KCl 40 mmol/L without or with a calcium blocker PN 200-100 (10^–6 mol/L); angiostein II (10^–11–10^–6 mol/L) without or with antagonist (losartan 10^–5 mol/L); or serotonin (10^–9–10^–4 mol/L) without or with antagonist (naftidrofuryl 10^–5 mol/L). Results thus obtained enabled us to propose a new hypothesis for the interpretation of the contractile responses of an elastic vascular smooth muscle. The different maximal effects of different agonists result mainly from the different proportions of cells they can mobilize; the agonist concentration-contraction relationship is mainly due to the increase of the proportion of cells involved up to a maximal value typical of the agonist used. An antagonist primarily reduce the proportion of cells an agonist can mobilize. Some of the consequences of this hypothesis are briefly outlined.     

TheChara plasmalemma at high pH. Electrical measurements show rapid specific passive uniport of H+ or OH−

Summary Above a critical external pH (about 10.5), theChara membrane acquires new propertes. In this state the membrane potential is close to the equilibrium potentials for H⁺ and OH^–, hyperpolarizing as external pH increases with a slope of –59 mV/pH unit. The membrane conductance increases by an average factor of 2.4 above the critical pH. These changes are explained by an increase in permeability to OH^– (or H⁺). The establishment of a OH^– (or H⁺ permeable membrane at high pH suggests that the large fluxes of OH^– (or H⁺ which occur in the alkaline band in photosynthesizing cells are passive.     

Pharmacological study of inhibited frog olfactory bulb glomerular input produced by a puff of air on the olfactory mucosa

The effects of applying 4-aminopyridine (10^–2 M), aminooxyacetic acid (AOAA — 10^–4–10^–3 M), -alanine (10^–3–10^–2 M), and bicuculline (10^–5, 10^–4 M) to the intact frog olfactory bulb were investigated. Having measured inhibition of orthodromic potential postsynaptic components produced either by a puff of air on the olfactory mucosa (OB input inhibition) or by single electrical stimulation of the olfactory nerve (postsynaptic inhibition) or by single electrical stimulation of the olfactory nerve (postsynaptic inhibition), it was found that 4-aminopyridine greatly intensified postsynaptic inhibition but strongly reduced that of OB input; inhibition of the latter was raised by AOAA or bicuculline and decreased by -alanine. These substances failed to exert any consistent, clear-cut effects on postsynaptic inhibition. Findings would support the hypothesis that OB input inhibition produced by a puff of air on the olfactory mucosa could occur as a result of GABA release from glial cells and subsequent binding of GABA to presynaptic GABA_B-receptors in glomeruli.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 19, No. 1, pp. 12–20, January–February, 1987.     

Characterization of H+/OH− currents in phospholipid vesicles

In pure phospholipid vesicles, the conductivity of H⁺/OH^– ions exceeds that for other simple inorganic ions. Protons achieve electrochemical equilibrium across egg phosphatidylcholine vesicles within tens of minutes. When pH gradients are established across vesicles, transmembrane potentials develop. Conversely, the establishment of transmembrane potentials leads to the formation of pH gradients. When the phenomenological permeability of H⁺/OH^– ions in vesicles is estimated, values are obtained that are much greater (six orders of magnitude larger) than those for Na⁺ or K⁺. A wide range in the values for this permeability has been reported; however, much of the discrepancy can be attributed to differences in the vesicle systems and experimental conditions. The H⁺/OH^– current appears to be modulated by changes in membrane dielectric constant. However, the dependence of this current on the pH gradient and on the membrane voltage argues against simple diffusion mechanisms as the source of the H⁺/OH^– current. In addition, in vesicle systems the H⁺/OH^– current shows a surprising invariance to changes in the membrane dipole potential, an observation that argues against the role of simple carriers for H⁺ and OH^– ions.     

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.