Dynamic defensive and feeding reactions to the acquisition of sensitization in edible snails]

In snails changes of defensive and alimentary behaviour and of reactions of command and modulator neurones of these kinds of behaviour were studied during elaboration of sensitization. After a single action of 50% chinine solution on mollusc's head, a short-term (during 50-70 min) and long-term (hours or days) facilitation was found of animals defensive reactions and responses of command neurones of defensive behaviour to tactile and chemical sensory stimuli. Alimentary behaviour of the snails and reactions of modulator neurones of alimentary behaviour to carrot juice presentation were inhibited in sensitized animals. Dynamics differences of defensive responses to tactile and chemical stimuli in the same sensitized animals were observed. Short-term responses facilitation under sensitization correlated chiefly with depolarization of the membrane potential of defensive behaviour neurones and with an increase of plasmatic membrane excitability. Prolonged facilitation of responses was due mainly to a change of effectiveness of synaptic transmission. The described model of sensitization elaboration can be a base for studying of molecular-cellular mechanisms, underlying the learning.     

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.     

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.     

Cellular reactions to elaboration of a backward conditioned connection in Helix lucorum

After 10-15 food stimuli paired with electrical shock in semi-intact snail preparation, responses to strong tactile stimuli identified feeding behaviour neurones were studied. Inhibition evoked by tactile stimulation in these cells before learning procedure disappeared and in some cases noxious stimulus evoked synaptic activation corresponding to feeding reactions in the intact animal. Changes in second-order sensory neurones pre-synaptic to the command neurones of avoidance behaviour are suggested to be the mechanism of forward conditioned connection as well as the mechanism of backward conditioned connection.     

Dynamics of changes in the excitability of Helix lucorum neurons in response to an analog of vasopressin

Application of desglycine-arginine-vasopressin to spontaneously nonactive command neurones of the snail's defensive reflex in the process of low-frequency (2-4 min intervals) intracellular stimulation led under certain conditions to an increase of excitability that was expressed in the shortening of latency of action potential generation, increase of the number of action potentials in response to a stimulus of fixed value, increase of membrane resistance, lowering of critical level of membrane depolarization and amplifying of pacemaker activity. However in spite of unidirectional changes of all the recorded parameters, the increases of values, opposite to the latency, did not correlate with the increases of membrane resistance and correlated well with the changes of depolarization critical level. If desglycine arginine-vasopressin was added to the medium during worsening of the neurones' excitability was probably caused by the influence of desglycine-arginine-vasopressin on the membrane active properties.     

Interrelationship between grape snail alimentary and defense behavior command neurons]

Food stimulus (carrot juice), releasing feeding behaviour in intact snails, evoked spike discharges in giant meta-erebral cells (considered to be command neurones of feeding behaviour) and subthreshold EPSPs in giant parietal cells (command neurones of avoidance behaviour) of a half-intact preparation. Tactile stimulation, eliciting avoidance reactions in intact snails, evoked hyperpolarization in command neurones of feeding behaviour and a spike discharge in command neurones of avoidance behaviour. Spikes induced into either of command neurones produced no changes in activity of other command cells. Inhibition of command neurones of feeding behaviour is assumed to be the basis of behavioural choice when food and tactile stimulus are presented simultaneously.     

Changes in the configuration of conditioned reactions of visual cortex neurons during changes in reinforcement parameters]

In order to study the influences of controlled changes of defensive integration on the activity of visual cortical units their responses to a conditioned light flash and electric cutaneous stimulation with a 600 msec interval between them were recorded in experiments on alert rabbits. It has been shown that in a third of the neurones the types of reaction to light flashes and electric stimuli coincide. The changes in parameters of the reinforcing shock led to a changed response of most cells to the conditioned photic stimulus and electric stimulation. The changes may have affected units which produce any activation phase, including cells with activity characteristic of detectory ("simple" and "complex") visual neurones. The data obtained suggest that the special function of the visual cortex is used in different ways in systemic mechanisms of conditioned and unconditioned defensive acts and that the integrated system of a behavioral act exerts control both on the use of the unit in a certain systemic process and on its receptive field.     

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.     

Neuronal activity of the cerebral cortex in cats with conditioned inhibition

Unit activity of the somatosensory cortex was studied in chronic experiments on cats during alimentary instrumental conditioned reflex performance and elaboration of conditioned inhibition (CI). First presentations of the CI signal at the beginning of CI formation caused, due to orienting reaction to a new stimulus, a levelling of the response of the neurones to the positive conditioned signal included into the CI complex. At the stage of consolidated CI, this depression proceeded gradually during the development of conditioned inhibition caused by consecutive presentations of the nonreinforced CI combination. Two groups of neurones were involved in the process of the CI performance: one of them being the same which was activated also in response to positive stimulation during performance of the conditioned response, and the second one being areactive to the positive conditioned stimulus but reducing the frequency of the background activity.     

Postsynaptic mechanism of withdrawal reflex sensitization in the snail

Repeated tactile stimulation of the skin evoked sensitization and subsequent habituation of a behavioral avoidance reaction of pneumostome closure in the land snail, Helix lucorum L. Spike responses in putative command neurons for avoidance behavior increased and subsequently decreased parallel to behavioral events. Decrease of spike responses in command neurons was due to decreased synaptic input evoked by repeated tactile stimulation. Intracellular investigation of the increase in the spike response in command neurons for pneumostome closure, which underlies behavioral sensitization, revealed a synpatically-evoked, long-lasting depolarization. No conductance changes during this depolarization were found, but neuronal excitability was altered.     

The neuronal organization of a focus of conditioned excitation in the cat sensory-motor cortex]

The activity of single units in the focus of conditioned excitation was studied during defensive conditioning to direct electrical stimulation of the cat sensorimotor cortex. Reorganizations of spike activity set in during the period of reflex elaboration, were manifest in the increased number of excited neurones and those which respond both to the conditioned and unconditioned stimuli. In the course of elaboration, the inhibitory phase of unit responses to direct electrical stimulation of the sensorimotor cortex was reduced, while the frequency of background unit spike activity was enhanced. Acute extinction of the reflex restored the initial duration of the inhibitory phase and reduced the frequency of the background activity.     

Excitability cycles of direct cortical responses during elaboration of conditioned defense reflexes in dogs]

Direct cortical responses (DCRs) to paired stimuli were studied in chronic experiments in dogs during elaboration of classical and instrumental defensive conditioned reflexes. The DCRs were recorded with 20 to 250 ms intervals between stimuli. Paired and single electrical stimulations of the middle suprasylvian gyrus given with a frequency of one per second were used as conditioned stimuli and were reinforced in a similar way. During electrical cutaneous stimulation of the dog's paw and to an even greater extent during isolated action of the conditioned stimulus the initial negativity of the testing DCR became shorter and the degree of its depression diminished. In the case of a following period of facilitation, its degree became greater. It was higher at a distance of 4 to 5 mm from the point of stimulation than at a distance of 2 to 3 mm. During isolated action of the conditioned stimulus, the degree of facilitation was higher than at the period of the possible action of the unconditioned stimulus. The greatest shorterning of the DCR excitability cycle was observed immediately before and during the conditioned lifting of the dog's paw. Excitability cycles of DCR, and possibly of other evoked potentials as well, are a more sensitive indicator of the function state of the cerebral cortex than responses to single stimuli. For this reason it appears promising to use them in studying conditioned reflexes.     

Effect of preliminary presentation of a future conditioned stimulus on the rate of elaboration of an instrumental defensive reflex in the rat

Preliminary presentation of future conditioned stimulus without reinforcement delayed and then accelerated subsequent elaboration of defensive conditioned reflex to this stimulus in rats. Preliminary presentation of resembling stimuli influenced the speed of conditioning in a lesser degree. It is suggested that such changes are due to formation of a local conditioned reflex to the future conditioned stimulus.     

The possibility of elaborating conditioned reflexes on the basis of emotional reactions evoked by electrical stimulation of limbic structures]

In experiments on cats with chronically implanted electrodes a conditioned stimulus (200 c/s tone) was paired with electrical stimulation of limbic structures, which produces pronounced emotional behaviour: defensive, aggressive, alimentary or drinking reactions. It has been shown that conditioned reactions are not elaborated on the basis of electrical stimulation, which activates alimentary, drinking or aggressive behaviour. Neither is a conditioned reflex formed to the combination of sound and stimulation of the brain structures, which inhibits alimentary behaviour. A distinct conditioned fear reaction sets in as a result of combination of sound and electrical stimulation of brain structures, which evoke defensive behaviour. The data obtained are analyzed in the light of the significance of natural requirements of the organism for the elaboration of conditioned reflexes.     

Characteristics of cortical responses to electrostimulation of the internal geniculate body during formation of a defensive conditioned reflex]

During elaboration of a classical defensive conditioned reflex the dogs exhibited a dependence of the changes in amplitude and configuration of evoked potentials (EP) to electrical stimulation of the medial geniculate body (MGB), a conditioned stimulus, on the nature of effector manifestation of the conditioned reflex: the late components were the most depressed at multiple phasic reactions and not infrequently increased and became complicated at single and short motor reactions as well as at their incidental absence. The primary oscillations, while mostly remaining unchanged, were depressed in the case of conditioned reactions attended with a general motor restlessness. A difference has been revealed during conditioning in the EP changes to electrical stimulation of MGB and to an adequate peripheral stimulation. It has been assumed that EP changes during conditioned activity are determined by the relationship between the levels of tonic and phasic cortical activation.     

Effect of serotonin on the activity of the neurons involved in the realization of the avoidance reflex of the snail

Bath application of 10(-5) mol/l of serotonin (5-HT) elicited a 50% increase of summary EPSPs recorded in command neurones for avoidance behaviour. No significant changes of rest potential and input resistance were seen in these cells. 5-HT evoked an increase of spontaneous level of firing in motoneurones involved in the same reflex, as well as an increase in the number of spikes which paralleled increase of EPSPs to the same stimulus in command neurones. In sensory cells, presynaptic to the command neurones, application of 5-HT evoked a significant increase of excitability and of input resistance. Monosynaptic EPSPs recorded in the command neurones showed a 40% increase after serotonin application. It is concluded that the major locus of plastic changes evoked by 5-HT application in the neuronal chain underlying avoidance reflex is the synaptic contact between sensory and command neurones.     

The role of the lateral and medial hypothalamus in reproducing a motor reaction as the signal during the acquisition of classical conditioned reflexes]

In experiments on three dogs there was shown that testing electrostimulation of the lateral hypothalamus reproduced the motor reaction which is a signal stimulus at elaboration of classic alimentary conditioned reflexes (CRs) and did not reproduce it at elaboration of classic defensive CR. Testing electrostimulation of medial parts of the hypothalamus reproduced, as LH electrostimulation the "signal" motor reaction, but in less percentage of cases, during elaboration of classic alimentary CRs and did not reproduce it at elaboration of classic defensive CRs. The reproduction of the signal motor reaction at LH electrostimulation is connected with activation of backward conditioned connection from motivation structures of the hypothalamus to representation of the signal stimulus in the motor cortex.     

Dynamics of the activity of rabbit visual cortex neurons during elaboration of elementary conditioned reflexes to time and association

Elaboration of a reflex to time (with omissions of stimuli) and of association; light--light + electrocutaneous stimulation of the paw was studied on neurones of the rabbit visual cortex. It was shown that by similarity of patterns of the discharge to a conditioned stimulus and to reinforcement, 83% of neurones reveal the formation of a reflex to time, and 57%--elaboration of association. Three ways of coding plastic reorganizations of the neuronal discharges are suggested; by the total number of spikes in the response, by the total sum of inhibitory periods, and by the pattern of the response. It has been found that in the reflex to time the early components of the response to the stimulus are reproduced during omissions; in elaboration of the association the late components of the discharge reaction to reinforcement are mainly repeated. Most cells with plastic reorganizations possessed polysensority and revealed discharge sensitization during habituation.     

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.