Ionic mechanisms underlying modulating effects of serotonin on acetylcholine-induced responses in Helix pomatia neurons

The ionic mechanisms underlying modulatory effects of serotonin on acetylcholine-response in identified and nonidentifiedHelix pomatia neurons were investigated using voltage-clamping techniques at the neuronal membrane. External application of 10^–5–10^–4 M serotonin to the membrane of neurons responding to application of acetylcholine depending on Na⁺ depolarization (D_Na response) reduced membrane conductivity during response to acetylcholine without changing reversal potential of acetylcholine-induced current. Acetylcholine (10^–6–10^–4 M) administration took place 1–3 min later. Neurons with response to acetylcholine application dependent on Cl⁺ depolarization (D_Cl response) or hyperpolarization (H_Cl response) behaved similarly. Analogous effects could be produced by external application of theophylline which, together with the latency and residual effect characteristic of serotonin action points to the participation of intracellular processes associated with the cellular cyclase system in the changes produced by serotonin in acetylcholineinduced response. Serotonin brought about a shift in reversal potential and an increase in the acetylcholine-induced current in those neurons where this response was associated with changed permeability at the membrane to certain types of ions. During two-stage acetylcholine-induced response of the D_Na-H_K type, serotonin inhibited the inward current stage. Mechanisms underlying modulatory serotonin action on acetylcholine-induced response in test neurons are discussed in the light of our findings.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 20, No. 1, pp. 57–64, January–February, 1988.     

Response induced in prog motoneurons by glutamate application

It was shown during experiments on isolated frog brain (fromRana ridibunda) that response to microelectrophoretically injected glutamate on to various points on the somatodendritic motoneuronal membrane (GLU response) displayed the same properties as EPSP induced in the same motoneuron by activation of three different synaptic inputs. Techniques of transmembrane polarization and current chop by means of a single microelectrode were used in this research. Mean levels of reversal potentials of GLU response and EPSP occurring as a result of stimulating the reticular formation, dorsal root, and microstimulation of presynaptic elements at the point of glutamate application equaled –16.9 ± 1.7 (n=13), –6.8 ± 1.7 (n=13), –9.8 ± 1.8 (n=6), and –15.1 ± 1.4 mV (n=13), respectively. Summation of GLU response and EPSP were quasilinear. Changes (upwards) in conductance associated with GLU response did not exceed 10%. Findings would indicate that glutamate, acting on the postsynaptic membrane receptors, induces depolarization and may serve as transmitter in all three inputs investigated.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 20, No. 6, pp. 776–785, November–December, 1988.     

Ionic mechanisms of the fast (nicotinic) phase of the acetylcholine response of identified Planorbarius corneus neurons

Responses to electrophoretic application of acetylcholine and suberyldicholine were investigated in identified neurons (LPed-2 and LPed-3) isolated from the left pedal ganglion ofPlanorbarius corneus. When microelectrodes filled with potassium chloride were used the reversal potentials of responses to acetylcholine and suberyldicholine were less negative than when microelectrodes filled with potassium sulfate were used; these reversal potentials were shifted toward depolarization if chloride ions in the medium were replaced by sulfate. These facts indicate that the responses in both LPed-2 and LPed-3 depend on chloride ions. Reversal potentials for acetylcholine and suberyldicholine in LPed-3 were virtually identical (–51 and –50 mV respectively), but in LPed-2 they differed significantly (–46 and –62 mV respectively). Replacement of sodium ions by Tris ions shifted the reversal potential for acetylcholine in LPed-2 toward hyperpolarization but did not change the reversal potential for suberyldicholine. Benzohexonium had the same action. The reversal potential for acetylcholine in medium with a reduced sodium concentration or in the presence of benzohexonium was the same as for suberyldicholine. It is concluded that on neuron LPed-2 acetylcholine activates both acetylcholine receptors which control conductance for chloride ions and acetylcholine receptors which change conductance for sodium ions, whereas suberyldicholine acts only on acetylcholine receptors responsible for the chloride conductance of the membrane.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 533–540, September–October, 1980.     

Properties of the slow excitatory postsynaptic potential in mammalian sympathetic ganglion cells

Two types of slow excitatory postsynaptic potentials (EPSPs) with different properties were found in neurons of the rabbit superior cervical sympathetic ganglion. In our group of neurons slow EPSPs increased during artificial hyperpolarization and decreased during depolarization of the membrane. The input resistance of the cells fell or remained unchanged during the development of slow EPSPs. In the second group of cells slow EPSPs increased during depolarization and decreased during hyperpolarization. The reversal potential of these responses, determined by extrapolation, was –78.9±3.6 mV. Depolarization responses to activation of muscarinic cholinergic receptors by acetylcholine or carbachol developed in 53% of neurons with an increase in input resistance and had a reversal potential of –83.2±6.7 mV. It is suggested that in cells of the first group the ionic mechanism of the slow EPSPs is similar to that of the fast EPSPs, whereas in cells of the second group its main component is a decrease in the potassium conductance of the membrane.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 13, No. 4, pp. 371–379, July–August, 1981.     

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.     

Blocking action of Nephila clavata spider toxin on ionic currents activated by glutamate and its agonists in isolated hippocampal neurons

The blocking action ofNephila clavata spider neurotoxin, or JSTX, on ionic currents activated by L-glutamate and its agonists when applied to the membrane of neurons isolated from the rat hippocampus was investigated using a concentration clamp technique. Crude JSTX venom was found to block L-glutamate-, quisqualate, and kainate-activated ionic currents induced by activating non-N-methyl-D-aspartate (non-NMDA) membrane receptors. Following the effects of JSTX, ionic currents activated by L-glutamate and its agonists declined to 34–36% of their initial value with no recovery during JSTX washout. An active fraction of JSTX at concentrations of 10^–4–10^–5 produced almost total but partially reversible blockade of ionic currents. The action of JSTX became less effective during depolarization. The concentration dependence of JSTX-induced blockade of kainate-activated ionic currents was investigated and the velocity constants of interaction between the toxin and glutamate receptors obtained. It is postulated that JSTX interacts with chemically-operated non-NMDA ionic channels, blocking their transition into a number of their possible open states.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 152–160, March–April, 1989.     

Ionic mechanisms of excitatory action of transmitter,exogenous acetylcholine,and serotonin on superior cervical ganglion neurons in rabbits

Ionic mechanisms of EPSP generation and depolarization induced by iontophoretic application of acetylcholine (ACh) and serotonin (5-hydroxytryptamine, 5-HT) — acetylcholine and serotonin potentials — were investigated in neurons of the isolated rabbit superior cervical ganglion by means of intracellular microelectrodes. The reversal potentials (E_r) for EPSP and the ACh-potential were –14.4±1.6 and –16.5±1.2 mV respectively, and they were about the same for the 5-HT potential. In some neurons (about one-third) much more negative values for E_r were obtained for EPSP and the ACh-potential by extrapolation, probably due to an increase in the resistance of their membrane during hyperpolarization. A decrease in the external sodium and potassium concentrations was shown to make E_r for EPSP and the ACh-potential more negative, whereas an increase in the external potassium concentration made it more positive than in normal solution; a change in the external chloride concentration did not alter E_r. It is suggested that the excitatory transmitter and exogenous ACh (and also, probably, 5-HT) share the same ionic mechanism of action of the membrane, which includes an increase in the permeability of the membrane to two ions — sodium and potassium — simultaneously.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 10, No. 6, pp. 637–644, November–December, 1978.     

Effects of applying oxytocin to Helix pomatia neurons. II. Hyperpolarization

The ionic mechanisms of hyperpolarization produced by applying oxytocin (OT) were investigated at the membrane of identifiedHelix pomatia neurons. Two types of neuron were known to exist, in one of which hyperpolarization is produced by a reduction in chloride ions at the membrane and a rise in membrane permeability to potassium ions in the other. In the first of these, response to OT had a reversal potential of –40 mV and decreased when furosemide and tolbutamide were added to the external medium. In the second case, the potential of the reversal of the response to OT was –70 mV. Upon doubling of potassium ion concentration in the external solution it was shifted towards depolarization by 15 mV. It is sugested thatHelix pomatia neurons have different types of OT receptors, some of which, when activated, manifest reduced chloride permeability at the membrane (probably through the cell cyclase system) with a rise in potassium permeability at the membrane in others.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 20, No. 5, pp. 659–666, September–October, 1988.     

Argiopine,argiopinines, and pseudoargiopinines as glutamate receptor blockers in hippocampal neurons

A homologous set of low-molecular weight compounds selectively blocking ionic currents were purified from venom from the spiderArgiope lobata with a selective blocking action on ionic currents activated by applying glutamate and its agonist kainic acid (KA) to the membrane of neurons isolated from the rat hippocampus. Three groups of these compounds — argiopine, argiopinines, and pseudoargiopinines, produced voltage-dependent glutamate- and KA-activated ionic currents at concentrations of 10^–6-10^–4 M, interacting primarily with agonist-activated ionic channels without affecting K_d values of the agonist. The blocking action could be partially reversed by argiopine application but only slightly when argiopinines and pseudoargiopinines were used. Kinetics of toxin effects on Ka-activated ionic currents showed at least two exponential components with different time constants. Simple and reversed rate constants of interaction between toxins and ionic channels were estimated from the plot of the kinetics of ionic current blockade and recovery against toxin concentration. Argiopine, argiopinines, and pseudoargiopinines lend themselves to further research into glutamate receptors of the mammalian CNS employing electrophysiological and biochemical techniques.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev, M. M. Shemyakin Institute of Bioorganic Chemistry, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 21, No. 6, pp. 748–756, November–December, 1989.     

Action of lead on glutamate-activated chloride currents inHelix pomatia L. neurons

Summary 1. In molluscan neurons glutamate may, on different neurons, evoke either excitation or inhibition. We studied neurons ofHelix pomatia which have hyperpolarizing responses to glutamate and determined the effects of lead on these responses.2. In voltage clamp experiments, the reversal potentials of these glutamate responses indicate that they are due to a conductance increase to chloride ions. Further evidence for this conclusion was obtained by the demonstration that responses to glutamate remained unaffected in experiments with intracellular dialysis with K-free saline in the presence of Na- and K-free extracellular media. In these circumstances, there is effectively no other ion than chloride to carry the current. In isolated neurons the glutamate-evoked chloride current is concentration dependent between 25 and 2500 µM. The current rises over 200 msec and declines in the continued presence of glutamate over a period of about 3 sec.3. Lead (0.5–1.0 µM) potentiated the glutamate-evoked chloride current provided that the channels were not maximally activated. The potentiation was greater if lead was added 30–60 sec before glutamate application.4. These results suggest that potentiation of transmitter-evoked responses by lead must be considered as yet another possible site of action of lead on neurons, and thus this effect must be considered as a part of the mechanism responsible for the neurotoxicity of this heavy metal.     

Modulatory effects of oxytocin on functional properties of three types of cholinoreceptors in molluskan neurons

It was found that applying 10^–8 M oxytocin (OT) affects the functional properties of three types of cholinoreceptors under conditions of voltage clamping at the membrane of identified ganglia neurons inHelix pomatia. This neuropeptide depressed acetycholine-(ACh-)induced sodium-potassium-calcium current in neuron RB3 without altering reversal potential of ACh-induced current. Two (sub-) types of cholinoreceptors were distinguished on the basis of findings on OT effects on ACh-induced chloride currents; ACh-induced chloride current was reduced by the action of OT on the cholinoreceptors of one of these (neuron F4) and increased in the case of neurons D5 and F86. The effects of applying OT and serotonin were reversible but not cumulative. Injection of OT exerted an action on ACh-induced chloride current independent of that of OT application. Involvement of cyclic adenosine monophosphate in OT-induced bimodal modulation of functional properties of three types of cholinoreceptors was demonstrated.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziology, Vol. 22, No. 1, pp. 87–93, January–February, 1990.     

Characteristics of apple snail giant neurons capable of generating action potentials in sodium-free solutions

The ability of apple snail giant neurons to generate action potentials in solutions that lack sodium ions is associated with the input resistance of these neurons in such a way that the higher the input resistance is, the more pronounced is this ability. Neurons in which this ability is well expressed usually exhibit low resting potential values and a slow repolarization phase. When calcium ions are replaced with barium ions, the neurons retain their excitability in a sodium-free medium for a longer period of time. Raising the calcium ion concentration to 30 µmole may exert a restorative effect on neurons that have lost their excitability in a solution that originally lacked sodium ions but contained 10 µmole of calcium ions. Increasing the calcium ion concentration to 60 µmole leads to loss of excitability, which under these conditions can be restored by means of depolarizing the neuron with an outward current. The results are discussed from the point of view of the theory of ionic conductivity of the surface membrane of neurons. It is hypothesized that the ability of the surface membrane of neurons to make use of sodium or calcium ions in generation of action potentials depends upon its permeability to potassium ions.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 2, No. 1, pp. 100–106, January–February, 1970.     

Characteristics of electrical responses by the medial geniculate bodies in Vespertilionidae and Rhinolophidae to ultrasonic stimuli with different frequencies

The total electrical responses and action potentials of the neurons in the medial geniculate bodies in Vespertilionidae and Rhinolophidae were investigated. Maximum sensitivity to ultrasonic stimuli was recorded inMyotis oxygnathus (Vespertilionidae) in the range 10–40 kc/sec and 65–80 kc/sec, and in Rhinolophidae in the ranges 10–70 kc/sec and 81–86 kc/sec. Low thresholds were observed inMyotis oxygnathus for the frequencies covered by their echo-location cries, whereas the thresholds recorded in Rhinolophidae in the 80 kc/sec band (the principal frequency of their echo location cries) were 15–30 dB higher than those for adjacent frequencies. Minimum thresholds of off-responses were observed inMyotis oxygnathus in the range 50–60 kc/sec, and in Rhinolophidae in the range 78–80 kc/sec. The regions of neuronal response in both species of bat were generally similar in form to those of responses recorded in the medial geniculate bodies of other mammals. Some of the neurons in Rhinolophidae with a characteristic frequency of about 80 kc/sec were also sensitive to stimuli with one-half and one-third of the principal frequency. In Rhinolophidae the greatest selectivity for frequencies was possessed by neurons that responded within the range from 80 to 90 kc/sec.A. A. Zhadanov Leningrad State University. Translated from Neirofiziologiya, Vol. 3, No. 2, pp. 138–144, March–April, 1971.     

Effects of arginine-vasopressin and its derivatives on Helix pomatia cerebral neurons

Response to application of and superfusion with solutions containing arginine-vasopressin and its derivatives (VPS), was investigated in identifiedHelix pomatia neurons. Different VPS exerted a similar effect on neurons in all cases. De- and hyperpolarizing as well as modulatory effects were shown. Depolarizing and hyperpolarizing response was accompanied by a rise and fall in steady-state conductance of the cell membrane. Reversal potential of response was determined as in the region of chloride reversal potential. Adding furosemide to the extracellular solution reversibly inhibited response to VPS. It was concluded from this that both de- and hyperpolarizing response took the form of an increase in the amplitude of trans-membrane ionic current induced by injecting cAMP into the neuron under the effects of superfusing the preparation with a VPS-containing VPS solution. Specific VPS receptors, probably associated with the cell cyclic nucleotide system, are thought to exist at the membrane of someHelix pomatia neurons.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 3, pp. 368–373, May–June, 1990.     

Serotonin-induced changes in the action of functionally distinct neurons in Helix pomatia

The effects of serotonin on the amplitude of summated EPSP in interneurons and on the duration of action potentials in sensory neurons, interneurons, and motoneurons involved in avoidance behavior were investigated in functionally distinct neurons isolated from theHelix pomatia nervous system. The duration of action potentials in sensory neurons was found to increase under the effects of serotonin (and this could underly the rise in EPSP amplitude), although that of interneuron and motoneuron spikes did not change. The functional significance of selective neuronal response to a rise in serotonin concentration is discussed, together with the mechanics underlying such effects.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 19, No. 3, pp. 316–322, May–June, 1987.     

Effects of applying oxytocin to Helix pomatia neurons. I. Depolarization

Inward current produced by applying oxytocin (OT) to the neuronal soma (OT_I) current) under conditions of voltage-clamping at the cell membrane was investigated inHelix pomatia. Replacing sodium with Tris ions in the external medium produced a considerable decline in OT_I current. A reduction in the external concentration of chlorine ions by replacement with HEPES ions induced an increase in OT_I current and a shift in its current-voltage relationship towards depolarization values. The presence of furosemide in the external solution reversibly inhibited OT_I current. This current likewise declined reversibly following external application of imidazole and tolbutamide but was increased by theophylline action. It was inferred that OT receptors are present on the surface membrane of someHelix neurons which, when activated, lead to increased chlorine permeability — a process apparently mediated via the cyclic nucleotide system.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR. Kiev. Translated from Neirofiziologiya, Vol. 20, No. 5, pp. 652–659, September–October, 1988.     

Ryanodine inhibits calcium transients evoked by action potentials in a subpopulation of rat dorsal root ganglion neurons

Effects of ryanodine on calcium transients evoked by depolarization of external membrane under voltage clamp conditions or by a train of action potentials under current clamp conditions were studied on isolated dorsal root ganglion neurons of newborn rats. In 70% neurons tested, ryanodine, a blocker of Ca²⁺-induced Ca²⁺ release from endoplasmic reticulum, significantly decreased the amplitude of calcium transients. The data obtained indicate that the Ca²⁺-induced Ca²⁺ release plays an important role for calcium signal generation in a subpopulation of sensory neurons.Neirofiziologiya/Neurophysiology, Vol. 26, No. 6, pp. 420–422, November–December, 1994.     

Unit responses of the ventral cochlear nucleus in bats of the rhinolophidae family to ultrasonic stimuli after destruction of the ipsilateral cochlea

Investigation of single unit responses in the ventral cochlear nucleus of the Rhinolophidae to ultrasonic stimuli after destruction of the ipsilateral cochlea revealed two groups of neurons with latent periods of: 1) 2–4 msec and 2) 5–32 msec. The first group has responses of low thresholds confined to narrow regions of the spectrum, the second has responses with high thresholds in wide regions. Neurons of the second group are also characterized by small changes in latent period and number of action potentials in response to a change in stimulus strength, large changes in threshold at characteristic frequencies depending on the stimulus duration, but only slight dependence of the thresholds on the time of the increase in strength. The pathways of activation of these neurons and their functional role are discussed.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 1, No. 1, pp. 32–40, January–February, 1972.     

Corticofugal postsynaptic influences on red nucleus neurons

The responses of red nucleus neurons to stimulation of the sensorimotor cortex was studied on nembutal-anesthetized cats. Most of the rubrospinal neurons were identified according to their antidromic activation. Stimulation of the sensorimotor cortex was shown to evoke in the red nucleus neurons monosynaptic excitatory potentials with a latency of 1.85 msec, polysynaptic excitatory potentials (EPSP), and inhibitory postsynaptic potentials (IPSP) with a latency of 9–24 msec. The EPSP often produced spikes. The probability of generation of spreading excitation is greater with motor cortex stimulation. The monosynaptic EPSP are assumed to arise under the influence of the impulses arriving over the corticorubral neurons as a result of excitation of axodendritic synapses. The radial type of branching of red nucleus neurons facilitates the transition from electrotonically spreading local depolarization to an action potential triggered by the initial axonal segment. Polysynaptic EPSP and IPSP seem to be a result of activation of fast pyramidal neurons whose axon collaterals are connected via interneurons with the soma of the red nucleus neurons.L. A. Orbeli Institute of Physiology of the Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 3, No. 1, pp. 43–51, January–February, 1971.     

Volume-dependent Glutamate Permeation Depends on Transmembrane Ionic Strength and Extracellular Cl<Superscript>−</Superscript>

Many mammalian cells regulate their volume by the osmotic movement of water directed by anion and cation flux. Ubiquitous volume-dependent anion currents permit cells to recover volume after swelling in response to a hypotonic environment. This study addressed competition between glutamate (Glu) and Cl^– permeation in volume-activated anion currents in order to provide insight into the ionic requirements for volume regulation, volume-dependent anion channel activity and to the architecture of the channel pore. The effect of changing the intracellular molar fraction (MF) of Glu and Cl^– on conductance and relative anion permeability was evaluated as a function of the extracellular permeant anion and/or the ionic strength. Relative permeability of Glu to Cl^– was determined by measuring reversal potentials under defined ionic conditions. Under conditions with high (150 mM) or low (50 mM) ionic strength solutions on both sides of the membrane, Cl^– was always more permeable than Glu. When a transmembrane ionic strength gradient (150 mM extracellular: 50 mM intracellular) was set to drive water into the cell, and in the presence of extracellular Cl^–, Glu became up to 16-fold more permeable than Cl^–. Replacement of extracellular Cl^– with Glu abolished this effect. These results indicate that it is possible for Glu to move into the extracellular environment during volume-regulatory events and they support the emerging role of glutamate as a modulator of anion channel activity.