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.     

Electrophysiological and neurochemical study of long-term sensitization in the snail

The long-term sensitization of avoidance reflex was produced in snail Helix pomatia, which led to the remarkable increase in the pneumostome closures period. The formation of long-term sensitization is also accompanied by increase in excitability of command neurons of this reflex. One of the possible mechanisms of this phenomenon is the depolarization of these cells. The quantitative redistribution of water-soluble proteins with relative mobility 0.54 0.42 0.40 was also observed in the identified neurons, both included in the avoidance reflex (command neurons) and non-included (bursting neurons, nerve cells of pool D). The protein with the relative mobility of 0.75 was unique for the nerve cells of neurosecretory pool D in sensitized snails, and was never found in control animals.     

Dynamics of elaboration of a conditioned reflex and differentiations in the snail

It is shown that dynamics of percentage of conditioned food refusals by snails Helix pomatia and Helix lucorum is satisfactorily approximated by regression equation of exponential type with included coefficient reflecting the animals state before the beginning of learning. The ability is established of the snails to practically completely differentiate various alimentary conditioned stimuli. The introduction of differentiation always improved the reflex consolidation. Conditioned reactions to differentiation stimulus appeared at the elaboration stage and were absent at the stage of conditioned reflex consolidation.     

Modulation of the Aplysia gill withdrawal reflex by dopamine

The ability of dopamine to modulate gill contractions was tested in Aplysia. When dopamine was perfused through the gill vasculature, gill contractions caused by siphon stimulation (gill withdrawal reflex) and by depolarization of the gill motor neuron L7 were increased in amplitude, as compared with those evoked during seawater perfusion. Habituation of gill movements, brought about by repetitive stimulation of the siphon or of L7, was prevented by dopamine. Despite the absence of reflex habituation, the number of action potentials in central gill motor neurons, evoked by siphon stimulation, showed normal decrement. Dopamine's effects were blocked when the ctenidial nerve was cut or when L7 hyperpolarized. These data suggest that dopamine acts peripherally to increase the efficacy of L7's synaptic transmission onto gill muscle or elements of the gill neural plexus.     

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.     

Contribution of serotonin to establishing a conditioned food aversion reflex in the snail

Combined presentation of food and noxious electrical stimulation produced no response in snails injected 6–16 days previously with 5,7-dihydrooxytryptamine, which produces degeneration of serotoninergic nerve terminals and reduced serotonin synthesis, although a defense (aversive) response was observed in the control group. Application of serotonin to a preparation of the snail central nervous system (contained in a bath) was used as reinforcement during neurophysiological experiments. The amplitude of synaptic response to nerve stimulation increased significantly in preparations in which stimulation was paired with serotonin application. After 3–7 sets of twin stimuli consisting of serotonin application and applying a drop of juice to the chemoreceptive surface area of the skin, a spike response to the latter stimulus was produced. No such effects were seen in response to unpaired stimuli. It was deduced that serotonin makes a major contribution to establishing conditioned aversive reactions in the snail.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 18, No. 3, pp. 291–298, May–June, 1986.     

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.     

Conditioning and sensitization in the edible snail: the neurophysiological and metabolic characteristics]

Electrophysiological parameters and bound calcium (Ca(b)) level dynamics during sensitization development or conditioning of food aversion were studied in the command neurons of defense behaviour in the snail Helix lucorum. Responses evoked by a testing sensory stimulus were facilitated 50-60 min after the first sensitizing stimulation, while conditioned responses appeared 80-90 min after the first conditioning. It was the most essential electrophysiological difference between the long-term sensitization and conditioning. Analysis of the Ca(b)) dynamics in the neurons showed significant differences in calcium-dependent metabolism during the sensitization and conditioning, likely underlying the electrophysiological differences.     

Sex differences in nociceptive withdrawal reflex and pain perception

Experimentally induced pain often reveals sex differences, with higher pain sensitivity in females. The degree of differences has been shown to depend on the stimulation and assessment methods. Since sex differences in pain develop anywhere along the physiological and psychological components of the nociceptive system, we intended to compare the nociceptive flexion reflex (NFR) as a more physiological (spinal) aspect of pain procession to the verbal pain report of intensity and unpleasantness as the more psychological (cortical) aspect. Twenty female and twenty male healthy university students were investigated by use of nociceptive flexion reflex threshold (staircase method) after electrical stimulation of the N. suralis. Furthermore, we assessed supra-threshold reflex responses (latency, amplitude and area) by applying 10 stimuli 5 mA above reflex threshold. Following each stimulation, the subjects provided pain ratings of intensity and unpleasantness on a visual analogue scale. Females exhibited marked lower nociceptive flexion reflex thresholds than males, while the supra-threshold reflex response tailored to the individual reflex threshold did not show any significant differences. The verbal pain ratings, corrected for NFR threshold, were not found to differ significantly. The large sex differences in nociception that were present in NFR threshold but not in the pain ratings corroborate the hypothesis that spinal processes contribute substantially to sex differences in pain procession.     

Effect of 6-hydroxydopamine on the electrical characteristics of snail neurons in long-term sensitization

Studies of the influence of neurotoxin 6-hydroxydopamine selectively destroying the catecholamine terminals on long-term sensitization, and the role of dopamine in manifestation of characteristics of a membrane of identified neurons during elaboration of plasticity, were fulfilled. Injection of saline was used as the control. It is shown that preliminary injection of 6-hydroxydopamine reduces duration of long-term sensitization, but does not block it completely. It was shown that injection of 6-hydroxydopamine prevents diminishing of membrane and threshold potentials in withdrawal interneurons during formation of long-term sensitization. The experiments demonstrate that shift of electrical characteristics of withdrawal interneurons caused by injection of neurotoxin 6-hydroxydopamine to both naive snails and sensitized snails, statys during at least 10 days.     

Context dependency of a limb withdrawal reflex in the caterpillar Manduca sexta

The proleg withdrawal reflex in the caterpillar Manduca sexta is a robust, well-characterized system for investigating the integration of sensory information with centrally patterned behavior. The reflex is evoked by stimulating mechanosensory hairs--planta hairs--located at the tip of each proleg. We studied the expression of this reflex by combining video recordings and electromyographic recordings from the main retractor muscles of the proleg, the principal and accessory planta retractor muscles. In intact animals, the nature of the response depended on the motor context of the animal. Animals which were standing quietly showed great variability in both the kinematic properties of proleg withdrawal, and the corresponding muscle electrical activity. Animals which were hanging upside down from a wooden dowel exhibited a much faster reflex, with retraction of the proleg occurring slightly faster than in standing animals, but re-extension of the proleg to the substrate being considerably faster. In crawling animals, expression of the reflex depended on the phase of the crawling cycle during which stimulation occurred. The reflex in a given proleg was suppressed during stance phase of that proleg. During swing phase, however, planta hair stimulation evoked proleg withdrawal, resulting in an assistance reflex. In contrast. isolated abdomens showed much less variability in the reflex. A comparison of the relationship between retractor muscle activity and the resulting proleg movement showed significant correlations between both the duration of activity and the number of muscle spikes, and the size of the associated proleg withdrawal. This is a promising system in which to investigate how central neuronal circuits accomplish context-dependency of motor behavior.     

L9 modulation of gill withdrawal reflex habituation in Aplysia

Repeated tactile stimulation of the siphon in Aplysia normally results in habituation of the gill withdrawal reflex and a concomitant decrease in the amplitude of the excitatory synaptic input to gill motor neurons in the abdominal ganglion. It was found, however, that induced low-level tonic activity in motor neuron L₉, which does not itself elicit a gill withdrawal movement, prevented habituation of the reflex from occurring. Further, in preparations already habituated, this tonic low-level activity brought about a reversal of habituation. Although tonic L₉ activity prevented the occurrence of habituation or brought about its reversal, it did not interfere with the synaptic decremental process which normally accompanies gill reflex habituation. Motor neurons L₇ and LDG₁ were found not to possess this ability of L₉ to modulate gill reflex habituation. Evidence suggests that L₉'s modulatory effect is mediated in the periphery, in the gill and not centrally in the abdominal ganglion.     

Sensitization inHelix snail: Morphofunctional correlates in command neurons of withdrawal behavior

Electrophysiological effects have been studied in command neurons of withdrawal behavior inHelix snail. In parallel, correlated changes in the content of bound calcium (Ca-b), as well as changes in DNA condensation, were investigated using a chlortetracycline fluorescent probe and the fluorescent dye bisbenzimide, respectively. Short-term electrophysiological changes (depolarization of the membrane and elevation of its excitability) in sensitized snails have been found to be accompanied by an increase in the Ca-b level in the cell nucleus and by partial DNA decondensation. Long-term effects were characterized by more pronounced synaptic components of the responses — slow EPSPs evoked by sensory stimuli, as well as by further DNA decondensation and considerable elevation of the Ca-b content in the nucleus and cytoplasm. The results obtained inin vitro conditions have shown that Ca-binding nonhistone proteins of chromatin are components of the cell nucleus whose content may be measured by chlortetracycline fluorescence.Translated from Neirofiziologiya, Vol. 25, No. 2, pp. 150–157, March–April, 1993.     

Antagonists of NMDA glutamate receptors selectively affect synaptic mechanisms of the nociceptive sensitization in the snail

The effects of N-methyl-D-aspartate (NMDA) glutamate receptor antagonists on the mechanisms of nociceptive sensitization were studied in LPl1 and RPl1 neurons of the semiintact preparation of a Helix lucorum snail. Application of sensitizing stimuli on the head part of the control preparation led to a depolarization of the membrane and increase in its excitability. A depression of responses of neurons evoked by tactile or chemical sensory stimulation during the short-term period and significant facilitation of responses during the long-term period of sensitization were observed. Sensitization performed under conditions of application of NMDA antagonists (AP5 or MK801) produced similar changes in membrane potential, membrane excitability, and neuronal responses evoked by tactile stimulation of the head or foot. However, the chemical stimulation of the head under these conditions evoked a significant depression of responses during the short- and long-term sensitization periods. The results suggest that the NMDA glutamate receptor antagonists selectively affect the plasticity induction mechanisms of the command neuron synaptic inputs, which mediate the chemical sensory stimulation from the snail's head.     

Suppression of gill withdrawal reflex behaviours in Aplysia: the role of acetylcholine

Acetylcholine (ACh) dissolved in seawater and perfused through the isolated gill of the Aplysia californica produced suppression of the gill withdrawal reflex (GWR) evoked by tactile stimulation of the gill. This suppression was reversible upon washout and was blocked by co-perfusion of curare and alpha-bungarotoxin. Co-perfusion of atropine did not block the suppression of the GWR produced by ACh. We concluded that the suppressive effects produced by perfusion of ACh through the gill occur as a result of the action of ACh at the nicotinic-like receptors. The role of ACh suppression in the mediation of gill reflex behaviours is discussed.