Possible relation between learning and non-template RNA synthesis in neurons

A substrate of either matrix-(cytidinetriphosphate, CTPh) or non-matrix (cytidinediphosphate, CDPh, or uridinediphosphate, UDPh) RNA synthesis was injected into mollusc neurone by means of intracellular microionophoresis. Subsequently, habituation was elaborated, assuming that if this process depends on the RNA synthesis, then the formation of defective RNA molecules with an excessive content of cytidine or uridine will disturb the learning process. In control experiments, habituation was elaborated either without drugs administration or following an injection of cytidinemonophosphate non subject to polymerization, or of an equimolar mixture of four ribonucleotidediphosphates: CDPh+UDPh+ADPh++GDPh. In the latter case a minimal distortion of the ratio between the initial substrates of the non-matrix RNA synthesis was expected. Both in the control experiments and following CTPh administration, the habituation was attended with an increase of threshold and latencies of spikes evoked by repeated stimuli. Injection of CDPh or UDPh disturbed the dynamics of thresholds and latencies increase in the course of habituation. The data obtained suggest that the habituation process is connected with a non-matrix synthesis of RNA in the neurones.     

Mollusk neuron habituation following extra- and intracellular administration of an ACTH fragment

By means of intracellular recording of the mollusc neuronal activity a study was made of the influence of ACTH4-7--a fragment of the stress hormone,--on elaboration of habituation to tactile stimuli. ACTH was administered immediately after the elaboration of the series I of habituation (25 to 30 stimuli) by means of either perfusion or extra- or intracellular microionophoresis. This was followed by the II-V habituation series (with 15 min intervals between them). Perfusion and extracellular (not intracellular) ACTH4-7 administration produced an acceleration of habituation in the series II, increase of action potentials (AP) thresholds in the response to the first tactile stimulus of series II and cessation of further increase of the AP thresholds in the course of elaboration. Without ACTH, these phenomena were observed beginning with habituation series IV. The ACTH action while influencing the memory is apparently directed to the external side of the excitable neuronal membrane; ACTH seems to accelerate the consolidation process. Any of the methods of ACTH administration produced also an enhancement of EPSPs and an increase in the number of APs in the responses to stimuli, without lowering the thresholds of APs generation. This testifies toi enhanced chemosensitivity of the membrane and may underlie the influence of ACTH on the level of wakefulness.     

Sensitization and habituation of the swimming behavior in ascidian larvae to light

Ascidian larvae of Ciona intestinalis change their photic behavior during the course of development. Newly hatched larvae show no response to a light stimulus at any intensity. At 4 hr after hatching, larvae were induced to start to swimming upon the cessation of illumination, and to stop swimming upon the onset of illumination. At a weaker light intensity (5.0 x 10(-3) J/m (2).s), the larvae showed similar responses to either a single stimulus or repeated stimuli of onset and cessation of light until 10 hr after hatching. At a stronger light intensity (3.2 x 10(-1) J/m(2).s), when the stimulus was repeated, they showed sensitization and habituation of the swimming response. At 3 hr after hatching the larvae failed to show any response to an initial stimulus at any intensity of light, but after several repeated stimuli (sensitization) they showed a swimming response at light intensities above 4.0 x 10(-2) J/m (2).s. At 5 hr and with intensity above 1.0 x 10 (-2) J/m(2).s, the larvae showed photoresponses to the first stimulus, but after several repetitions the larvae failed to stop swimming upon the onset of light (habituation). A repeated series of stimuli at stronger intensities of light caused greater habituation; this habituation was retained for about 1 min. Since the larval central nervous system in Ciona is comprised of only about 100 neurons, learning behavior in ascidian larvae should provide insights for a minimal mechanism of memory in vertebrates.     

Photoresponse and Learning Behavior of Ascidian Larvae, a Primitive Chordate, to Repeated Stimuli of Step-Up and Step-Down of Light

Ascidians are lower chordates and their simple tadpole-like larvae share a basic body plan with vertebrates. Newly hatched larvae show no response to a stimulus of light. 4 h after hatching, the larvae were induced to swim upon a step-down of light and stop swimming upon a step-up of light. At weaker intensity of light, the larvae show the same response to a stimulus after presentation of repeated stimuli. When intensity of actinic light was increased, the larvae show sensitization and habituation of the swimming response to a stimulus after repeated stimuli of step-down and step-up of the light. Between 2 h 20 min and 3 h 40 min after hatching the larvae did not show any response to the first stimulus, but after several repeatedstimuli they show swimming response to a step-down of light. A repeated series of stimulus cause sensitization. Between 4 h and 7 h after hatching, the larvae show photoresponse to the first stimulus, but after several repetition of the stimuli, the larvae could not stop swimming to a stimulus of a step-up of the actinic light. A repeated series of stimulus cause greaterhabituation. Both sensitization and habituation depend upon intensity ofactinic light.     

Signaling via the FLP-14/FRPR-19 neuropeptide pathway sustains nociceptive response to repeated noxious stimuli in C. elegans

In order to thrive in constantly changing environments, animals must adaptively respond to threatening events. Noxious stimuli are not only processed according to their absolute intensity, but also to their context. Adaptation processes can cause animals to habituate at different rates and degrees in response to permanent or repeated stimuli. Here, we used a forward genetic approach in Caenorhabditis elegans to identify a neuropeptidergic pathway, essential to prevent fast habituation and maintain robust withdrawal responses to repeated noxious stimuli. This pathway involves the FRPR-19A and FRPR-19B G-protein coupled receptor isoforms produced from the frpr-19 gene by alternative splicing. Loss or overexpression of each or both isoforms can impair withdrawal responses caused by the optogenetic activation of the polymodal FLP nociceptor neuron. Furthermore, we identified FLP-8 and FLP-14 as FRPR-19 ligands in vitro. flp-14, but not flp-8, was essential to promote withdrawal response and is part of the same genetic pathway as frpr-19 in vivo. Expression and cell-specific rescue analyses suggest that FRPR-19 acts both in the FLP nociceptive neurons and downstream interneurons, whereas FLP-14 acts from interneurons. Importantly, genetic impairment of the FLP-14/FRPR-19 pathway accelerated the habituation to repeated FLP-specific optogenetic activation, as well as to repeated noxious heat and harsh touch stimuli. Collectively, our data suggest that well-adjusted neuromodulation via the FLP-14/FRPR-19 pathway contributes to promote nociceptive signals in C. elegans and counteracts habituation processes that otherwise tend to rapidly reduce aversive responses to repeated noxious stimuli.     

Ventilation response thresholds do not change with age or self-reinforcement in workers of the bumble bee Bombus impatiens

The response threshold model is a potential mechanism for task allocation in social insects, and it assumes that workers vary in the levels of task stimuli to which they respond. Furthermore, response thresholds of individual workers may change over time through self-reinforcement (experience), such that workers become more sensitive to task stimuli. However, in addition to self-reinforcement, aging is another process that occurs through time. Distinguishing whether response thresholds change within workers due to self-reinforcement or aging may give insight into the flexibility of this task allocation mechanism. Using a ventilation paradigm, we manipulated workers of Bombus impatiens to have either repeated or lack of exposures to increases in nest air temperature, thereby allowing us to manipulate experience and thus self-reinforcement. Nest air temperature was the task stimulus, and ventilation (fanning) was the behavioral response. We found that ventilation response thresholds do not decrease either with age or experience in workers of B. impatiens, contrary to what has been reported for B. terrestris workers (Weidenmüller, 2004). Instead, we found high levels of intra-individual variation in response thresholds. Our results also show that workers with lower average response thresholds respond to heating events with higher probability than those with higher ventilation thresholds. These results provide insight into the role of the response threshold framework for task allocation; we also discuss how response probabilities may play a role in task allocation among workers.     

Skin potential and EMG changes induced by cutaneous electrical stimulation. I. Normal man in arousing and non-arousing environment.

Skin potential and EMG responses induced in normal man by electrical stimuli applied to the skin were recorded in the four limbs in order to study somato-sympathetic and somato-motor reflexes. Different patterns of responses were observed in different conditions: alarm, habituation, sensitization and arousal. During alarm, sensitization and arousal, the responses were present in the four limbs; during habituation, the responses were only present in the stimulated and in the contralateral limb. Three sensory thresholds to cutaneous electrical stimulation were identified in habituated subjects: tactile, tingling and pain. Cutaneous and EMG responses appeared at tingling threshold. A relationship between skin potential level and skin potential response was observed.     

Amplitude,latency, and habituation of the electrodermal response to acoustic stimuli in the frog

The electrodermal response (EDR) of frogs to various acoustic stimuli was measured in the form of the skin potential response (SPR). There was no correlation between the polarity of the SPR and the intensity of the stimuli. When different frequencies were presented at the same intensity, the amplitude of the SPR to each was inversely proportional to the sound pressure at which that frequency just elicited an SPR. The amplitude of the sound-induced SPR increased monotonically with increasing sound pressure. The latency of the SPR decreased with increasing intensity of the acoustic stimulus. Acoustic stimuli repeated at intervals of 1 and 2 min elicited responses with progressively decreased amplitude and increased latency; with 4 min intervals there was little habituation. Fatigue participates to only a very slight extent in reducing the amplitude of the SPR and increasing its latency. The results are compared with the published data on frogs and mammals, including humans.     

Habituation of an odorant-induced startle response in Drosophila

Habituation is a fundamental form of behavioral plasticity that permits organisms to ignore inconsequential stimuli. Here we describe the habituation of a locomotor response to ethanol and other odorants in Drosophila, measured by an automated high-throughput locomotor tracking system. Flies exhibit an immediate and transient startle response upon exposure to a novel odor. Surgical removal of the antennae, the fly's major olfactory organs, abolishes this startle response. With repeated discrete exposures to ethanol vapor, the startle response habituates. Habituation is reversible by a mechanical stimulus and is not due to the accumulation of ethanol in the organism, nor to non-specific mechanisms. Ablation or inactivation of the mushroom bodies, central brain structures involved in olfactory and courtship conditioning, results in decreased olfactory habituation. In addition, olfactory habituation to ethanol generalizes to odorants that activate separate olfactory receptors. Finally, habituation is impaired in rutabaga, an adenylyl cyclase mutant isolated based on a defect in olfactory associative learning. These data demonstrate that olfactory habituation operates, at least in part, through central mechanisms. This novel model of olfactory habituation in freely moving Drosophila provides a scalable method for studying the molecular and neural bases of this simple and ubiquitous form of learning.     

Dynamics of habituation in different cortical regions of the cat brain

The dynamics of habituation in the rostral part of the parietal association region and also in the first and second somatosensory areas was studied by the evoked potentials (EP) method in cats anesthetized with chloralose (80-90 mg/kg) and immobilized with flaxedil. During repeated stimulation of the splanchnic nerve brief habituation of EP developed in the cortex. It corresponded in its basic features to habituation observed in other sensory systems. On comparison of the rates of development of habituation in the primary and secondary projection zones of the splanchnic nerve in the cortex differences were found: habituation developed more rapidly in the secondary projection zones. The functional importance of this phenomenon is discussed.     

Characteristics of the neuronal activity of structures of the hippocampal formation (subiculum)]

Spontaneous and evoked neuronal activity of subiculum was investigated in chronic unanaesthetized rabbits. Rhythmic components of two different types were observed in the spontaneous activity of 58% of units: 4.5-6.0 cps bursts (13%) and 2.0-3.8 cps (45%). Besides their basic frequency, these types of rhythmic activity were differentiated by several additional criteria. 63% of the cells responded to applied sensory stimuli, among which auditory stimuli were the most effective. Multimodal units were slightly more numerous (59%) than unimodal ones. In 50% of the recorded units responses were of tonic type, in 31%--of phasic type, and in 19%--specific on-off responses were observed. In majority of units with repeated stimulations relatively rapid process of habituation developed after a short incremental phase. Interruption of septal afferents resulted in disappearence of 4.5-6.0 cps (theta) bursts, in reduction in the number of tonic responses and in absence of habituation. Similarity of neuronal characteristics in the field CA1 and subiculum is discussed.     

Inhibition of spinal interneuronal activity by repeated cutaneous stimulation: a possible substrate of flexor reflex habituation

The responses of interneurones, situated in the lumbar region of the rat spinal cord, to repeated cutaneous stimulation, were studied. The main purpose of this investigation was to attempt to determine the extent to which habituation of the flexor reflex might be explained on the basis of a progressive development of inhibition. Spontaneously active interneurones, which were inhibited by cutaneous stimuli, were investigated in detail. In rats whose spinal cords were intact, the period of inhibition was shown to increase with successive stimuli. This increase in inhibition was directly related to the intensity and frequency of stimulation, occurred more rapidly during a second series of stimuli and was antagonized by strychnine. In spinal animals, an increase in the duration of the period of inhibition to repeated stimulation could not be demonstrated. In this preparation, a gradual decrease in inhibition occurred. It is tentatively concluded that inhibition of spinal interneurones, the development of which depends upon descending influences from supraspinal centres, may be partially responsible for habituation.     

Dynamics of single unit responses of the rabbit visual cortex to repetitive photic stimulation

Single unit responses in the visual cortex of the waking rabbit to repetitive photic stimulation at a frequency of once every 2.5 sec were studied. Depending on the total number of spikes in the response, the dynamics of the responses could be divided into two types: "fast" and "slow." From 5 to 15 stimuli were required to establish a stable level of response with changes of the first type, but 50 to 100 stimuli were needed for the response with changes of the second type. About 50% of all neurons did not change the characteristics of response. In the group of neurons with changing responses, partial habituation was found in 55–59% of cells; there were 25% of neurons with sensitization of discharge and 17–20% with a humpbacked type of response dynamics. A "slow" dynamics of unit responses in most cases was accompanied by changes in the duration of inhibitory pauses in the response; negative correlation of these values was observed in 65% of neurons.M. V. Lomonosov Moscow State University. Translated from Neirofiziologiya, Vol. 10, No. 5, pp. 451–459, September–October, 1978.     

The neurochemical basis of recurrent inhibition in the reflex arch of the defensive reaction]

Mechanisms of habituation in the network of identified neurones were investigated in isolated preparation of central nervous system in the snail Helix. It has been found that intracellularly induced spike discharge in premotor command neurones decreases synaptic responses to repeated nerve stimulation in all recorded command neurones. Application of the neuropeptide FMRFamide elicits similar changes in the network. Taking into account that the investigated command neurones contain FMRFamide, as was shown immunochemically, it is possible to assume the existence of recurrent inhibition in the network underlying avoidance reactions. This recurrent inhibition causes habituation of the network output in the cases when the repeated stimuli do not evoke sensitization via activation of serotonergic cells.     

Giant clams discriminate threats along a risk gradient and display varying habituation rates to different stimuli

It is often beneficial for animals to discriminate between different threats and to habituate to repeated exposures of benign stimuli. While much is known about risk perception in vertebrates and some invertebrates, risk perception in marine invertebrates is less extensively studied. One method to study risk perception is to habituate animals to a series of exposures to one stimulus, and then present a novel stimulus to test if it transfers habituation. Transfer of habituation is seen as a continued decrease in response while lack of transfer is seen either by having a similar or greater magnitude response. We asked whether giant clams (Tridacna maxima) discriminate between biologically relevant types of threats along a risk gradient. Giant clams retract their mantle and close their shell upon detecting a threat. While closed, they neither feed nor photosynthesize, and prior work has shown that the cost of being closed increases as the duration of their response increases. We recorded a clam's latency to emerge after simulated threats chosen to represent a risk gradient: exposure to a small shading event, a medium shading event, a large shading event (chosen to simulate fish swimming above them), tapping on their shell and touching their mantle (chosen to simulate different degrees of direct attack). Although these stimuli are initially perceived as threatening, we expected clams to habituate to them because they are ultimately non‐damaging and it would be costly for clams to remain closed for extended periods of time when there is no threat present. Clams had different initial latencies to emerge and different habituation rates to these treatments, and they did not transfer habituation to higher risk stimuli and to some lower risk stimuli. These results suggest that clams discriminated between these stimuli along a risk gradient and the lack of habituation transfer shows that the new stimulus was perceived as a potential threat. This study demonstrates that sessile bivalves can discern between levels of predatory threat. These photosynthetic clams may benefit from being able to categorize predator cues for efficient energy allocation.     

The neuronal reactivity and interaction of the rat cerebral cortex during the microiontophoretic action of acetylcholine in a model negative learning situation]

Unit activity of 46 pairs of neurons of sensorimotor cortex of rats was studied in a model situation of habituation to repetitive microiontophoretic applications of acetylcholine (ACh). The difference between the types of reactions to ACh of two neighbouring neurons recorded by the same microelectrode was observed on 37% of cases. The difference between the dynamics of activity of neurons with similar patterns of reactions during repetitive action of stimuli was also shown. Stability of the excitatory connections between two neighbouring neurons under the conditions when one of them demonstrated the habituation to repetitive action of ACh were indicated by analysis of cross-correlation histograms.     

Feedback control and quantification of the response of EEG alpha to visual stimulation

The response of the posterior EEG alpha rhythms to visual stimulation is so variable that it is difficult to obtain reliable on-line measurement of it. Feedback between the EEG alpha and the visual stimulus (1) reduces random variation in the response and (2) facilitates on-line quantification. With feedback EEG, the response to visual stimulation is measured as a series of time durations of alpha and of no-alpha intervals in the EEG. This time series occurs in two stages: an initial disturbance followed by a recovery. The quantification of the series of time durations is achieved by fitting curves to the series of alpha time intervals and of no-alpha time intervals. These functions, computed in each trial of 30 stimulations, are an objective, quantitative definition of EEG response. The utility of the method was demonstrated by testing it with reference to well-known effects. Habituation to a repeated stimulus, dishabituation, habituation to a class of stimuli, dishabituation by changing the class of stimuli, and differences among brain-lesioned, psychiatric patients and normals were shown with a detailed quantification. It was concluded that biofeedback is the method of choice for quantitative research on the EEG component of the human orienting response.     

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.     

Neurophysiological mechanisms underlying habituation of the tentacle retraction reflex in the land slug Ariolimax

Habituation of the tentacle retraction reflex was studied at the following response levels: (1) Muscle tension elicited in the tentacle retractor muscle by repeated stimulation of a cerebral nerve (at 60-sec intervals) declined in parallel with evoked activity of the largest unit in the tentacle retractor nerve. (2) The largest unit in the tentacle retractor nerve (L₄) showed spontaneous recovery and dishabituation. The rate of response decrement was inversely related to the strength of stimulus, and an optimal interstimulus interval ca. 60 s was found. Retention of habituation for 24 h was exhibited. (3) The major retractor motoneurons (L₂, L₃, L₄) all showed habituation, dishabituation, and spontaneous recovery. The decline of L₄ activity was parallelled by a decline in muscle response. (4) Compound EPSPs elicited in the retractor motoneurons by stimulation of sensory pathways showed habituation and dishabituation. (5) Unitary EPSPs elicited by stimulation of cerebral nerves and connectives with minimal stimulus strengths also showed habituation and were unaffected by spontaneously occurring EPSPs. Dishabituation by another pathway was also shown. (6) Depolarization of L₄ by a constant current produced spike trains of constant firing rate and evoked a constant level of muscle tension in repeated trials, suggesting the absence of habituation in a peripheral nerve net or at the neuromuscular junction.