Electrophysiological study of serotoninergic neuron Cl in the pteropod mollusk Clione

Functional characteristics of cerebral serotoninergic neuron Cl, axons of which terminate at the buccal ganglia [7], were investigated in the pteropod molluskClione. Stimulating neuron Cl induced activation of the feeding rhythm generator located in the buccal ganglia — an effect arising after a long latency and persisting for some tens of seconds once stimulation had ended. Neuron Cl receives feedback from buccal ganglion cells and this brings about periodic modulation in ganglia activity during the generation of feeding rhythm. Activity of neuron Cl is correlated with operation of the locomotor rhythm generator located in the pedal ganglia. The firing rate of Cl neurons increased upon activation of the locomotor generator (whether spontaneous or induced by stimulating certain command neurons). The correlation found between workings of the locomotor generator and activity of Cl neurons is thought to be one of the manifestations of feeding synergy involving simultaneous activation of the locomotor and buccal apparatus.Institute for Research on Information Transmission, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 1, pp. 18–25, January–February, 1991.     

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.     

Regeneration of pedal ganglion neurons in the sea butterfly Clione limacina

In the pedal ganglia ofClione limacina the growth of neurites is traced in motoneurons after transection of the wing nerve and in interneurons after transection of the pedal commissure. Neurons were stained intracellularly with Lucifer yellow. In the motoneurons the neurites growing from the transected end of the axon and from the neuron soma spread to all nerve trunks departing from the ipsi- and contralateral ganglia. For nerve transection in the intact mollusk, wing movements were restored 10 days after the operation. In the interneurons the growing neurites branched within the pedal ganglion or spread to the cerebral ganglia, but they never reached the periphery.Institute of Problems of Information Transmission, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 17, No. 4, pp. 449–455, July–August, 1985.     

Neurons determining passive defensive response in the pteropod mollusk Clione limacina

Neurons responding to tactile stimulation of the head with bursts of action potentials of short latency followed by passive defensive response were found in the pedal ganglia and identified as Pd13. Stimulation of one Pd13 neuron leads to inhibition of the entire locomotor generator. A whole set of neurons, identified as P2, 3, 4, and 5, activated solely by intensive tactile stimulation of the head, were found in the pleural ganglia. Stimulating one such neuron also induces inhibition of the entire locomotor generator. These pleural cells are synaptically connected with Pd13 neurons and one EPSP in Pd13 unit corresponds to each action potential in the pleural cell. This connection has a facility for potentiation, subsequently replaced by habituation. In this way, pleural neurons also introduce Pd13 neurons into the inhibitory trend when activated by intensive tactile stimulation. Application of cerucal and ergotamine (dopaminergic receptor blockers) suppresses the inhibitory effect of the Pd13 neuron and pleural cells, thus indicating dopamine involvement in the inhibitory processes occurring in passive defensive reaction.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 21, No. 5, pp. 685–694, September–October, 1989.     

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.     

Neural control of heartbeat in the pteropod mollusk Clione limacina

The heart of the pteropod molluskClione limacina is innervated by the median nerve arising from the left abdominal ganglion. Five neurons sending axons to the heart have been identified in theClione central nervous system with retrograde cobalt or Lucifer yellow staining. Neuron H1 located in the left pedal ganglion produced an excitatory effect on heart beat. Stimulation of three neurons, H2–H4, situated in a compact group in the medial region of the left abdominal ganglion, led to inhibition of cardiac contraction, while H5, located in the caudal region of the left abdominal ganglion, did not affect heart beat. The activity of efferent cardiac neurons (ECN) was found to be related to the operation of the locomotor rhythm generator. Spontaneous or reflex depression of the latter was found to inhibit neuron H1 and activate units H2–H4. The behavior of these ECN accounts for the positive correlation between heart operation and locomotor activity inClione limacina.Institute of Research on Information Transmission, Academy of Sciences of the USSR, Moscow, M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 185–192, March–April, 1989.     

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.     

Chemical sensitivity of Limnaea stagnalis neurons and their responses to synaptic stimulation

Responses of nerve cells to puncture, to touching the surface of the mollusk leg, osmotic stimulation, and extracellular microiontophoretic injection of acetylcholine, noradrenalin, serotonin, atropine, and propranolol were recorded intracellularly in the right parietal, left pedal, and visceral ganglia of the unisolated circumpharyngeal ring ofLimnaea stagnalis. Selective sensitivity of the neurons to the biologically active substances was observed. Results indicative of the functional differences between the various ganglia and of their neurochemical organization were obtained. Selective blocking of the unit responses to puncture of the surface of the mollusk leg by atropine or propranolol suggests that different forms of excitation reaching the central neurons evoked different and specific neurochemical processes on their subsynaptic membranes which can retain the essential informativeness of the widely different afferent volleys converging on a single nerve cell.I. M. Sechenov First Moscow Medical Institute. Translated from Neirofiziologiya, Vol. 5, No. 5, pp. 510–518, September–October, 1973.     

Involvement of group S-100 brain specific proteins in the neurophysiological mechanisms of habituation

The possible role of S-100 brain specific proteins and their involvement in the neuronal mechanisms of habituation were investigated in identified neurons of the snail (Helix pomatia). Extracellular application and intracellular injection of antibodies into S-100 proteins caused a rapid and significant reduction of neuronal response to repeated stimulation compared with the control. A considerable reduction in the amplitude of evoked EPSP and an increase in the duration of latency of potentials was produced by the antibodies and numbers of action potentials declined. Meanwhile, membrane potential, membrane input impedance, and neuronal pacemaker potentials remained virtually unchanged. This effect is presumed to occur due to breakdown of synaptic transmission processes. The subject of possible involvement of S-100 proteins in the operation of synaptic structures as a primary apparatus for integrating neuronal activity is considered.P. K. Anokhin Institute of Normal and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow; Institute of Physiology, University of Novi-Sad, SFRYu. Translated from Neirofiziologiya, Vol. 19, No. 5, pp. 637–645, September–October, 1987.     

Neurochemical mechanisms of burst activity regulation in isolated snail endogenous oscillators: Role of monoamines in opoid peptides

The electrical activity of three identified bursting neurons (including one "plateau-generating" neuron) is studied in the brain ofHelix lucorum under conditions of complete isolation. A method for isolating the identified neurons is worked out (mechanical extraction by means of microelectrodes). All three neurons are shown to be endogenous oscillators. In the isolated state in the absence of any stimulation they exhibit spontaneous changes in activity: from an inactive state to nongrouped regular activity, from regular activity to burst activity, and vice versa. It is also found that the burst activity of all three isolated neurons can be regulated (initiated, discontinued, parameters altered) using the same neurochemical agents: serotonin, opioid enkephalins. Burst activity can be generated by the combined action of dopamine and enkephalin and discontinued just with dopamine. The possible significance of the described neurochemical mechanism of regulating burst activity in neurons that are endogenous oscillators is discussed with regard to the generation of the rhythm of various cyclic functions.N. K. Kol'tsov Institute of Developmental Biology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 472–480, July–August, 1991.     

Sites of action potential generation in Helix pomatia neurons

The site of action potential generation in unipolar snail neurons was identified by stimulating neurons isolated together with the initial portion of the process from the neuropile. Stimulation consisted of a sinusoidal from electrical current passed along the soma-axonal axis in saline solution. No low threshold sites of action potential generation were found in 80% of test neurons using this technique. Spontaneous activity was determined by the operation of one dominant site on the neuronal process. Antidromic activation of the soma by axonal action potentials (even with simultaneous hyperpolarization of the soma) induced somatic potentials more successfully than direct somatic depolarization by the current flowing through the solution.Institute of Chemical Physics, Academy of Sciences of the USSR, Moscow. Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 20, No. 1, pp. 90–98, January–February, 1988.     

Effect of intracellular injection of cyclic amp on electrical characteristics of identified neurons in Helix pomatia

The effect of intracellular iontophoretic injection of cyclic AMP on electrical activity of neurons RPa1, RPa3, LPa2, LPa3, and LPl1 in the corresponding ganglia ofHelix pomatia was investigated. Injection of cyclic AMP into neuron LPl1 was found to cause the appearance of rhythmic activity (if the neuron was originally "silent"), an increase in the frequency of spike generation (if the neuron had rhythmic activity), and a decrease in amplitude of waves of membrane potential, in the duration of the interval between bursts, and in the number of action potentials in the burst (if the neuron demonstrated bursting activity). In the remaining "silent" neurons injection of cyclic AMP led to membrane depolarization. Injection of cyclic AMP into neurons whose membrane potential was clamped at the resting potential level evoked the development of an inward transmembrane current (cyclic AMP current), the rate of rise and duration of which increased proportionally to the size and duration of the injection. Theophylline in a concentration of 1 mM led to an increase in the amplitude and duration of the cyclic AMP current by about 50%. It is concluded that a change in the cyclic AMP concentration within the nerve cell may modify the ionic permeability of its membrane and, correspondingly, its electrical activity.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 517–525, September–October, 1980.     

Responses of an identified Helix pomatia neuron to application of peptides,mediators, prostaglandins,and cyclic nucleotides

Extracellular application of oxytocin, Lys-vasopressin, and Leu-enkephalin to neuron RPa1 ofHelix pomatia evoked the generation of pacemaker potentials and the appearance of potentiation of spike activity of bursting type, characteristic of this cell. Noradrenalin and prostaglandins of the E group had a similar action. Dibutyryl-cyclic AMP, the phosphodiesterase inhibitor papaverine, and sodium fluoride, a nonspecific activator of adenylate cyclase, also initiated or potentiated bursting discharges of the neuron. It is suggested that the effects of oxytocin, Lys-vasopressin, Leu-enkephalin, noradrenalin, and prostaglandins of the E group are mediated through intracellular processes linked with activation of adenylate cyclase by these substances, leading to an increase in the cyclic AMP content in the nerve cell.P. K. Anokhin Research Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 13, No. 1, pp. 80–87, January–February, 1981.     

Neuronal mechanisms of defense reaction to cutaneous nerve stimulation

A two-stage defense response was induced in the freshwater snailPlanorbarius corneus by stimulating the head. It consisted of the shell being rapidly lowered over the head and foot followed by the snail gradually withdrawing into its shell. These movements are performed by contracting the columellar muscle. Motoneurons of the columellar muscle were identified in the cerebral, parietal, and pedal ganglia. Stimulating the lip nerve was found to induce 2-stage excitation in motoneurons (responsible for the 2-stage muscular contraction) in preparations of central nervous system with the columellar muscle attached. The same 2-stage motoneuronal excitation can also occur spontaneously. This implies that defense reaction in the cell is at least partially a "fixed action" underlying a central mechanism or program and triggered by afferent stimuli. Activation of the central mechanism of defense response can also induce depolarization in certain columellar muscle motoneurons. This points to the existence of a feedback between neurons of the central mechanism and motoneurons.Institute of Research on Transmission of Information, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 22, No. 6, pp. 786–795, November–December, 1990.     

Variability of adaptational properties of an excitable neuron membrane

This investigation was made on a preparation of stretch receptors of molting crayfish. We made intracellular recordings of potentials from the soma of a slowly-adapting neuron, and extracellular recordings from the nerve trunk. After strychnine had been added to the physiological solution surrounding the preparation, additional rhythmic activity was recorded from the nerve trunk, with corresponding depolarizational oscillations in the membrane potential of the soma of the slowly-adapting neuron. The additional rhythmic activity had a competitive relationship to the action potentials lying along the axon of the slowly-adapting neuron, the rhythm frequency increasing as the prolonged action potentials arose in the soma of that neuron. The depolarizational oscillations in the soma did not change sign as its membrane potential decreased. Analysis of the above phenomenon led to the conclusion that within the axon membrane of a slowly-adapting neuron there appears a section that spontaneously generates rhythmic action potentials. The results of the investigation indicate that there may be wide variations in the adaptational properties of the neuron membrane.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 1, No. 3, pp. 309–314, November–December, 1969.     

Principle of neuronal organization of defensive reflexes in mollusks

Spontaneous and evoked synaptic activity of command neurons for the defensive response of spiracle closing were studied by simultaneous intracellular recording of activity of several identified CNS neurons in snails. Comparison of monosynaptic EPSPs in command neurons evoked by discharges of presynaptic neurons with spontaneous synaptic potentials indicated that the central organization of the defensive reflex is in the form of a two-layered neuron net in which each neuron of the afferent layer possesses a local receptive field, but which overlaps with other afferent neurons. Each neuron of the afferent layer is connected with each neuron of the efferent layer by monosynaptic excitatory connections that differ in efficiency (maximal only with one neuron of the efferent layer). Both receptive fields of neurons of the afferent layer and "fields of efficiency of synaptic connections" are distributed according to the normal law. As a result of this organization the neuron net acquires a new quality: The action of different stimuli leads to the appearance of differently located "spatial excitation profiles" of efferent layer neurons even when this action of the stimulus occurs not at the center of the receptive field.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 16, No. 1, pp. 26–34, January-February, 1984.     

Parameters of a model of transmitter depletion in synapses of identified snail neurons

Elementary EPSPs arising in two different identified neurons of the parietal ganglion ofHelix pomatia were recorded after stimulation of the identified triggering neuron. Repetitive stimulation (0.1–1.5 Hz) led to low-frequency depression of EPSPs. By the use of known and modified models of transmitter depletion parameters characterizing storage, mobilization, breakdown, and liberation of transmitter were determined. The fraction of available pool (F) released in two different synapses of the same trigger neuron did not differ significantly. The available pool of transmitter (C) and the demobilization constant ( ) in synapses on the RPa3 neuron were 2–3 times higher, and mobilization (M) was 10 times higher than on the LPa2 neuron. Predictions of the depletion model showed deviations from the experimental data. A method of calculating consistently whatever law of change of F was adopted was devised. Absence of correlation between parameters F and C of the depletion model and binomial parameters p and n, calculated on the basis of the quantal hypothesis of synaptic transmission shows that this hypothesis and the transmitter depletion model describe different synaptic mechanisms.Brain Institute, Academy of Medical Sciences of the USSR, Moscow. Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 13, No. 1, pp. 88–97, January–February, 1981.     

Naloxone-dependent depression by morphine of responses of snail neurons to serotonin

Morphine, added to the extracellular solution in a concentration of 1·10^–5 M, quickly and reversibly weakens the depolarizing and hyperpolarizing responses of neurons of the snailHelix lucorum evoked by 1·10^–6 M serotonin. The inhibitory effect of morphine is completely abolished by the addition of naloxone (1·10^–5 M), suggesting that opiate receptors are involved in the process. Interaction between morphine and serotonin is noncompetitive in type, as is shown by the character of the dose-effect curves recorded during the action of serotonin before and after morphine application.Institute of Psychiatry, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 13, No. 6, pp. 589–593, November–December, 1981.     

Synaptic mechanisms controlling receptor unit activity in the stretch receptor—Abdominal ganglionic chain system of the crayfish

Inhibitory control over activity of the receptor neuron was investigated in a preparation of the stretch receptor and abdominal ganglionic chain in crayfishes. Potentials were recorded intracellularly from receptor neurons and neurons of the abdominal ganglion, and extracellularly from the dorsal roots. IPSPs appeared in the receptor neuron in response to stimulation of that same neuron or of the abdominal ganglionic chain. The relationship between spikes at the input and output of the inhibitory neuron varied over a wide range depending on the functional state of the neuron. A linear relationship was established between the time before appearance of the IPSP and the duration of the interspike interval of the slowly adapting neuron (SAN) and also between the firing rate of this and the inhibitory neurons during recurrent inhibition. Factors influencing the length of the interspike interval of the SAN on the appearance of an IPSP in it were investigated. It is postulated that summation of potentials evoked by spikes of the SAN and also of potentials evoked by spikes of that neuron, together with local processes evidently of endogenous nature takes place in the inhibitory neuron. IPSPs were recorded from two neurons resistant to strychnine and blocked by picrotoxin on the receptor neuron. The structural and functional organization of the individual elements in the chain of recurrent inhibition and inhibition evoked by stimulation of the abdominal ganglionic chain is discussed.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 5, No. 3, pp. 323–332, May–June, 1973.     

Interaction between sea anemone toxin BgTX8 and ionic channels of neurons isolated from mammals

The action of the toxin BgTX₈ separated from the sea actiniaBunodosoma granolifera on transient tetrodotoxin-sensitive sodium and outward potassium currents of units isolated from rat sensory ganglia was investigated using techniques of voltage clamping at the membrane and intracellular perfusion. It was found that BgTX₈ decelerates the inactivation kinetics but has little effect on activation kinetics of sodium current. At the same time, a 5–10% increase in the amplitude of inward current was often observed at holding potentials of about –100 to –120 mV at the membrane. The dissociation constant of the receptor-toxin equals 4×10^–6 M and is adequately described by Langmuir's isotherm. It was also established that intracellular perfusion of neurons with anemone toxin-containing solution leads to a reduction in the amplitude of sodium current and decelerates its inactivation process. Suppression of outward potassium current was also noted.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Institute of Brain Research, Academy of Sciences, Havana, Cuba. Translated from Neirofiziologiya, Vol. 20, No. 1, pp. 32–37, January–February, 1988.