The effect of a vasopressin analog on the reaction of the command neurons in the defensive behavior of the edible snail during nerve stimulation

On the level of snail command neurones of the defensive behaviour an increase was revealed of the amplitude of summary EPSP recorded in command neurones in response to nerve stimulation, an increase of probability of appearance of action potential in the reaction and, under certain conditions, a significant decrease of habituation speed at rhythmic (0.1 Hz) nerve stimulation against the background of peptide. The latter effect was found during comparison of groups of neurones--control neurones and those to which the peptide was presented before the first series of stimulation in the test, i. e. without preliminary elaboration of habituation. The decrease of the speed of habituation concerned both the amplitude of summary EPSP and the probability of action potential appearance in the reaction. All these changes against the background of vasopressin analogue may be the basis of the increase of spike reactions of command neurones of the defensive behaviour and thus the basis of the initiation or the increase of behavioural defensive reactions. The obtained effects were not protracted and took place only in the presence of the peptide in the extracellular medium.     

Effect of changes in extracellular calcium concentration on electrical characteristics of command neurons after defensive reflex conditioning in snail

Studies of electrical characteristics of command neurons of the defensive reflex have shown that membrane potential does not reliable differ for both naive and learned snails with changing of the calcium concentrations in external solution. With increasing of the concentrations of calcium ions it is observed the increases of threshold potential from 14 + 0.7 mV in 2.5 mM Ca2+ to 21.8 + 0.9 mV in 20 mM Ca2+ for naive snails. Threshold potential decreases from 16.8 + 0.6 mV (saline solution for snail--10 mM Ca2+) to 13.3 + 0.6 mV in 20 mM Ca2+ with increasing of calcium concentrations for learned snails, and under its reduction it is also decreased, but less remarkably, than for naive, to 11.8 + 0.8 mV in 2.5 mM Ca2+. On the same scheme it is changed a critical level of depolarization: for naive snails it decreases with increasing of the calcium concentrations, and for learned snails it increases either with increasing of the calcium concentrations, and so with its reduction.     

The effect of the small cardioactive peptide (b) on synaptic transmission efficiency and on the excitability of command neurons in the defensive behavior of the edible snail]

Small cardioactive peptide (b) (SCPb) application in concentration 5 X 10(-8) mol/l into the saline surrounding the snail CNS leads to an increase of amplitude of summate excitatory postsynaptic potential in the command neurones (CN) elicited by the intestinal nerve stimulation. Besides, SCPb causes an increase of excitability of the CN. Described effects can change the threshold of the neuronal net underlying avoidance behaviour. The possibility of integrative effects of peptides in low concentration on the behaviour is discussed.     

Involvement of intracellular calcium in sensitization of command neurons of defensive behavior in the edible snail (Helix lucorum)

Examinations carried out on command neurons of defensive behavior in the edible snail using electrophysiological methods and a chlortetracycline fluorescent probe revealed that a single sensitizing action alters electrical neuronal activity and the amount of bound calcium in the cells. An initial increase in the amount of bound calcium (the first 15–20 min after the sensitizing action) coincides in time with depolarization, enhancement of plasma membrane excitability, and a decrease of amplitude and duration of the excitatory postsynaptic potentials (EPSP) induced by sensory stimulations. Repeated pronounced increase in the bound calcium level develops 50–60 min after the sensitizing action and correlates with facilitation of neuronal responses to sensory stimuli. Alterations in the bound calcium level in command neurons of defensive behavior in the course of sensitization development differed in dynamics and direction from the previously described bound calcium shifts in the same cells in the course of habituation development.P. K. Anokhin Institute of Normal Physiology, Academy of Medical Sciences of the USSR Moscow. I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR Leningrad. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 418–427, July–August, 1991.     

The mono- and polysynaptic connections between identified neurons in the system of the passive defensive reflex in the edible snail]

A structure of synaptic connections between the identified sensory and giant command neurons of Helix lucorum was studied. It was found that EPSPs arising in the giant neuron as responses to single action potentials generation in sensory neuron consist of several monosynaptic and several polysynaptic components having different magnitude, latencies, and plasticity. The latencies of monosynaptic components are determined by different presynaptic terminals' lengths.     

Behavioral changes during the acquisition of long-term sensitization of the defensive reflex in the edible snail

Reaction of long-term sensitization (LTS) of defensive reflex was elaborated in the snail. It was accompanied by a considerable increase of the time of closing of pneumostome. The results show that LTS is preserved for more than two week and may be a model of long-term memory.     

Sensitization and habituation of command neurons during a defensive reflex in grape snails

All investigated neurones were classified into functional groups depending on their participation in unconditioned avoidance reflex of pneumostome closure. Habituation was produced by tactile stimuli applied with a frequency of 0.1 c/s. Spike responses of neurons participating in processing sensory information habituated gradually, while reactions of command neurones of avoidance behaviour became sensitized to the second or third stimulus of the series. Behavioral reaction was sensitized in parallel with command neurones in spite of habituation in other types of nerve cells. A conclusion is drawn that behavioral habituation is due to waning of responses in all the participating neurones, but the independnet process of sensitization is due to sensitization of command neurones. Probable mechanisms of neuronal sensitization are discussed.     

The defensive behavior of the edible snail]

Literary and experimental data are summarized on organization of defensive behaviour in Helix. Muscular and neuronal mechanisms participating in this form of behaviour are reviewed.     

Selectivity of opioid peptide effects on excitability and various sensory inputs in LPl1 and PPl1 command neurons participating in defensive behavior of the snail Helix lucorum

Opioid peptides effects on neural membrane as well as neural responses evoked by sensory stimuli with different modality and site of application, were investigated in L-RPII command neurones of defensive behaviour of semi-intact preparation in the land snail Helix lucorum. Met-enkephalin (10 uM) application onto the snail CNS increases membrane excitability and produces facilitation of neural responses evoked by quinine solution (0.5%) application onto snail head and depression of reactions evoked by tactile stimulation of the head. Met-enkephalin in dose of 0.1 uM initiates only a depression of neural responses evoked by tactile stimulation of the head. Leu-enkephalin (10 uM) application suppresses neural reactions evoked by tactile stimulation of the head. Membrane excitability and neural responses evoked by quinine application onto the snail head do not change after leu-enkephalin administration. Effects appear 10-20 min after initiation of the peptide application. Initial neural responses were observed 15-30 min after CNS washing with Ringer solution. In addition, facilitation of neural responses evoked by chemical stimulation of the snail head was found 30-50 min after leu-enkephalin washing. Peptides do not change neural responses evoked by tactile stimulation of the snail foot. Neural effects of peptides were prevented by simultaneous naloxon administration (50 uM). Experimental results show selective opioid peptides' effects on excitability and plasticity of L-RPII neural inputs with site- and modality-specifics.     

Effect of cyclic AMP on the excitation and response of command neurons responsible for the snail defensive behavior caused by sensory stimulation]

Extraneuronal application of db-cAMP or intraneuronal injection of cAMP were found to increase the neural membrane excitability and synaptic facilitation in neural responses to sensory stimulation of Lpl1 and Rpl1 neurones. The db-cAMP exerted no effects on neural responses to tactile stimulation of the snail foot or mantle. The intraneuronal injection of cAMP produced synaptic facilitation only in neural responses to quinine application to the snail head. The findings suggest the cAMP selective involvement in postsynaptic mechanisms of inducing the long-term facilitation transient stage.     

Presynaptic mechanisms for intensifying reactivity of command neurons in Helix pomatia defensive behavior under the effects of vasopressin analog

This study set out to investigate presynaptic mechanisms underlying reactivity of command neurons inHelix pomatia defensive behavior under the effects of desglycine-arginine vasopressin, a vasopressin analog. Peptide perfusion was found to intensify accumulated EPSP in command neurons during stimulation of the nerve without affecting the monosynaptic connection between primary sensory and command neurons. It was concluded that parallel pathways exist for the spread of excitation from the periphery to command neurons. Properties of such pathways are described and neurons with similar properties are identified. It is in these same cells that long-term changes in spontaneous EPSP induced by application of the peptide were noted.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 22, No. 6, pp. 723–730, November–December, 1990.     

Long-term potentiation in the neurons of the edible snail

A brief high-frequency stimulation of the anal nerve of the isolated nerve ring of snail Helix induced a pronounced increase in the amplitude of EPSPs, evoked in identified neurons of left parietal and visceral ganglions by low frequency (once in 5 min) stimulation of the same nerve. The amplitude of EPSP returned to the control level 30-120 min after tetanization. We called this effect long-term potentiation. A brief application of serotonin (10 microM) in the majority of neurons also induced lasting either 15-30 min or more than 2 hours facilitation of EPSP, evoked by anal nerve stimulation. Intracellular cAMP injections, being without effect on EPSP amplitude in many neurons, in certain neurons caused an increase in EPSP amplitude, lasting up to 30 min. It is suggested that the 3 factors shown to increase synaptic efficiency in molluscan neurons may have common mechanisms of action.     

Electrophysiological study of 5,6-dihydroxytryptamine effect on defensive reflex conditioning in the snail

The role of serotonin in expression of membrane properties of identified neurons was studied during defensive reflex conditioning using the neurotoxic analogue of serotonin 5,6-dihydroxytryptamine (5,6-DHT). The defensive reflex conditioning in snails was destroyed on the second day after second injection of 5,6-DHT. Through the 1st weeks after second injection of 5,6-DHT the snails were learned but worse than snails after injection of saline solution. This result shows the recovery of snail's learning ability within 2 weeks after the second injection of 5,6-DHT. It was found that injection of 5,6-DHT prevented the decrease of membrane and threshold potentials of command neurons during defensive reflex conditioning as compared with the snails injected with 5,6-DHT without learning.     

Effect of the opiate antagonist naloxone on the electrical activity of identified neurons in the edible snail

The opiate antagonist naloxone modifies the electric activity of some identified neurons of the Helix lucorum which have not been preliminary exposed to the effect of exogenous opioids. Some neurons are excited while others are inhibited by naloxone, and in both cases the reaction may have both a short and long latent period. The reactions depend on naloxone dose and become less expressed or are blocked when naloxone is administered together with the agonists of opiate receptor (morphine, D-Ala2, D-Leu5-enkephalin, bremazocine and beta-endorphin). Opioids alone do not produce any effect on neurons. The effect of naloxone on neurons is assumed to be a result of the elimination by the opiate antagonist of the tonic effect of endogenous opioids by their replacing on opiate receptors which are constantly stimulated by endogenous ligands.     

Desensitization of choline receptors of somatic membrane command neurons in the snail

Desensitization of somatic membrane receptors to rhythmic application of acetylcholine was studied in command neurons PPa2, PPa3, LPa2, and LPa3 of the snail. The results indicate that desensitization of choline receptors is a relatively simple, short-term phenomenon usually not accompanied by more complex, long-term intracellular reorganizations. With short intervals between applications, however, densensitization may lead to a significant decrease in the reaction of the postsynaptic neuron and display certain properties characteristic also for depression of the synaptic reaction. On this basis we assume that in habituation, postsynaptic receptors participate in the processes of short-term decrease in synaptic efficiency.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 17, No. 4, pp. 530–538, July–August, 1985.     

Regression analysis of an instrumental conditioned tentacular reflex in the edible snail

Regression analysis revealed the opportunity of approximation with exponential mathematical model of the learning curves of conditioned tentacle reflex. Retention of the reflex persisted for more than three weeks. There were some quantitative differences between conditioning of the right and the left tentacle. There was formation of the reflex in every session during spring period, but there was no retention between sessions. The conditioned tentacle reflex may be employed in neuropharmacological studies.     

The role of dopamine and serotonin in modulating the defensive behavior of the edible snail

Dopamine application in concentration of 10(-5)-10(-6) M into saline around the snail CNS leads to decrease of excitability of LPa7 neurone which is presynaptic in relation to defensive behaviour command neurones, and to decrease of amplitude of monosynaptic excitatory postsynaptic potential (EPSP) in the command neurones elicited by intracellular stimulation of LPa7 neurone. Besides, the dopamine causes a decrease of summated EPSP amplitude in the studied neurones in response to intestinal nerve stimulation (70% in average), a change of rest potential towards hyperpolarization for 6-8 mV, a reduction of the command neurones input resistance (20% in average). The described influences can lead to a general increase of the threshold of defensive system reaction to stimulation. Dopamine action on the defensive behaviour command neurones is significantly weakened in serotonine presence. Against the dopamine background, the efficiency of serotonine influence on the value of EPSP in command neurones in response to testing stimulus is reduced. According to the obtained data, a conclusion is made that interrelation of dopamine and serotonine concentrations can be a base for formation of behaviour choice in snail.     

Elementary and compound postsynaptic potentials in the defensive command neurons of Helix lucorum

The present communication concerns with the analysis of elementary and the compound excitatory postsynaptic potentials (eEPSPs and cEPSPs) recorded by intracellular microelectrode from an identified defensive command neuron of the snail Helix lucorum. The eEPSPs were evoked by single presynaptic action potentials (APs) elicited by cationic current injection into one of the identified sensory neurons synapsing on the respective command neuron. The cEPSPs were elicited by local brief tactile stimuli on the skin or internal organs. It was shown that the cEPSPs amplitudes depend mainly on the number of activated sensory neurons. Compound EPSPs depend also on frequency and the number of APs in the bursts occurring in a single neuron. Presynaptic APs having frequency 2-10 Hz evoke high frequency depression of that eEPSPs after an interval is followed by post-tetanic potentiation of single eEPSPs. Preceding stimulation of a pneumostom area facilitates the cEPSPs elicited by repeated stimulation of viscera. The eEPSPs from the same visceral area demonstrate no heterosynaptic facilitation in experiments with double parallel intracellular recording from responsive sensory and command neurons. The different types of the eEPSPs plasticity are discussed according to their contribution cEPSPs plastic changes.