Habituation versus sensory adaptation as the cause of reduced attraction following pulsed and constant sex pheromone pre-exposure in Trichoplusia ni

Trichoplusia ni males receiving a pulsed regime of sex pheromone pre-exposure later, in a wind tunnel, exhibited reduced upwind flight and close approach to a sex pheromone source compared to males receiving continuous pre-exposure. Electroantennogram (EAG) amplitudes from either pulsed or continuously pre-exposed males were significantly reduced from controls only during exposure. EAG amplitudes returned within one minute after exposure to levels not significantly different from controls, indicating that habituation, not sensory adaptation was probably the cause of reduced flights under the pulsed regime. Additionally, activity levels of males during each pulsed pre-exposure remained high compared to constant or no pre-exposure males, implying that pulsing was better at achieving central nervous system habituation because it avoided adapting the receptors.     

Elevation of pheromone response threshold in almond moth males pre-exposed to pheromone spray

Abstract. High percentages of naive Cadra cautella (Walker) (Lepidoptera: Pyralidae) males not pre-exposed to pheromone flew upwind to sources containing 50 ng (83%) and 500 ng (97%) of pheromone, but not to sources containing 5 μg (23%) and 50 μg (4%).Of the naive males that flew upwind in response to 50 ng sources, 67% located and landed on the source, whereas fewer than 19% of the naive males that flew upwind in response to higher doses located and landed on the sources.A 2-minute pre-exposure of C.cautella males to a spray cloud containing 50 ng, 500 ng, 5 μg or 50 μg of pheromone, induced shifts in response levels such that in wind-tunnel bioassays performed 1 h later, there was an increase in the doses that optimally elicited upwind flight and landing on the source that was proportional to the pre-exposure dose.Few of the pre-exposed males flew upwind to (10–43%) and landed on (0–33%) 50 ng sources, whereas they now perferentially flew upwind to(58–81% and 52–73%) and landed on (33–68% and 55–60%) pheromone sources of doses of 500 ng and 5 μg, respectively.Therefore pre-exposure to pheromone promoted a shift of threshold for response, and not an overall reduction in responsiveness to pheromone.     

Responsiveness to sucrose and habituation of the proboscis extension response in honey bees

In honey bees, complex behaviours such as associative learning correlate with responsiveness to sucrose. In these behaviours, the subjective evaluation of a sucrose stimulus influences the behavioural performance. Habituation is a well-known form of non-associative learning. In bees, the proboscis extension response can be habituated by repeatedly stimulating the antennae with a low sucrose concentration. A high sucrose concentration can dishabituate the response. This study tests whether habituation correlates with responsiveness to sucrose in bees of different behavioural states and in bees which are habituated with different sucrose concentrations. Habituation and dishabituation in newly emerged bees, 5-day-old bees and foragers strongly correlated with responsiveness to sucrose. Bees with high responsiveness to sucrose displayed a lower degree of habituation and showed greater dishabituation than bees with low responsiveness. The degree of habituation and dishabituation also depended on the concentration of the habituation stimulus. These experiments demonstrate for the first time in a non-associative learning paradigm that the subjective strength of a sucrose stimulus determines the behavioural performance. Non-associative learning shares this property with associative learning, which suggests that the two processes might rely on similar neural mechanisms.Abbreviations: GRS Gustatory response score - PER Proboscis extension response     

Habituation of sexual response in male Heliothis moths

It has been generally hypothesized that habituation mediates the effects of pheromone‐based disruption strategies used in the management of moth pests. The current study demonstrates that pheromone‐mediated sexual response in the tobacco budworm moth, Heliothis virescens (F.) (Lepidoptera: Noctuidae), can in fact be modulated by conditions consistent with the production of habituation. An ethogram was used to measure response strength in a wind tunnel experiment where male moths were allowed to respond freely to one of two different blends of female pheromone in 16 trials over 4 days. Post‐test measures were collected to investigate stimulus specificity and long‐term effects. In conditions appropriate to the formation of habituation, habituation will develop and disrupt male sexual response to female sex pheromone. Males repeatedly exposed to plumes of synthetic pheromone blends display a habituated response lasting up to 96 h. Habituation rate and spontaneous recovery of response strength are greater with less intense stimuli. Additionally, males habituated to one blend express no habituation of sexual response when exposed to a different blend. This indicates a high degree of stimulus specificity, which could facilitate outbreeding, and that moths attend to the configuration of the pheromone blend, not simply to its elements.     

Reduction in responsiveness of male light-brown apple moths, Epiphyas postvittana, to sex pheromone following pulsed pre-exposure to pheromone components

ABSTRACT. A combination of two sex pheromone compounds is required to elicit sexual response in males of Epiphyas postvittana. Sequential exposure to the two compounds failed to elicit sexual response in males, but a pre-exposure sequence of alternated pulses of the two compounds did habituate males to the same extent as did pulsed pre-exposures to the compounds combined.     

Pheromone-mediated behaviour of male lightbrown apple moth, Epiphyas postvittana, correlated with adaptation of pheromone receptors

ABSTRACT. Two pheromone components are required to elicit close-range precopulatory behaviour in male lightbrown apple moths, Epiphyas postvittana (Walker) (Lepidoptera). The receptor cells which respond to the major component (I) ( E )-11-tetradecen-1-yl acetate, have a fast disadaptation rate with recovery occurring within 5 s after stimulation, while the cells responding to the second component (II), ( E, E )-9, 11-tetradecadien-1-yl acetate, recover after approximately 300 s.
Studies on the behaviour of males in the laboratory show a close correlation between the duration of a memory effect, during which males will respond to compound I alone after receiving an initial exposure to I and II, and the time-course for disadaptation of component II-responding cells. These results suggest several possibilities for mechanisms of integration of sensory input by the CNS.     

Brief exposure to sensory cues elicits stimulus-nonspecific general sensitization in an insect

The effect of repeated exposure to sensory stimuli, with or without reward is well known to induce stimulus-specific modifications of behaviour, described as different forms of learning. In recent studies we showed that a brief single pre-exposure to the female-produced sex pheromone or even a predator sound can increase the behavioural and central nervous responses to this pheromone in males of the noctuid moth Spodoptera littoralis. To investigate if this increase in sensitivity might be restricted to the pheromone system or is a form of general sensitization, we studied here if a brief pre-exposure to stimuli of different modalities can reciprocally change behavioural and physiological responses to olfactory and gustatory stimuli. Olfactory and gustatory pre-exposure and subsequent behavioural tests were carried out to reveal possible intra- and cross-modal effects. Attraction to pheromone, monitored with a locomotion compensator, increased after exposure to olfactory and gustatory stimuli. Behavioural responses to sucrose, investigated using the proboscis extension reflex, increased equally after pre-exposure to olfactory and gustatory cues. Pheromone-specific neurons in the brain and antennal gustatory neurons did, however, not change their sensitivity after sucrose exposure. The observed intra- and reciprocal cross-modal effects of pre-exposure may represent a new form of stimulus-nonspecific general sensitization originating from modifications at higher sensory processing levels.     

Reduced antennal sensitivity, behavioural response, and attraction of male codling moths, Cydia pomonella, to their pheromone (E,E)-8,10-dodecadien-1-ol following various pre-exposure regimes

The effects of pre‐exposing male codling moths, Cydia pomonella (L.) (Lepidoptera: Tortricidae), to their pheromone (E,E)‐8,10‐dodecadien‐1‐ol (codlemone), in static and moving air, under laboratory and field conditions, on subsequent antennal sensitivity, behavioural responsiveness, and attraction to codlemone were investigated. In flight tunnel experiments, the percentage of moths wing fanning and taking flight were mostly unaffected, but upwind flight to, and contact with, pheromone sources known to elicit responses of both were shown to depend on the intensity and duration of previous exposure to codlemone and recovery time between exposure and assessment. Ten to 30‐min pre‐exposures to codlemone in static air (≈ 35 µg l^?1) not only caused a 99% reduction in attraction, but also significantly reduced electroantennogram response to codlemone. Recovery of full antennal sensitivity to codlemone took more than 1 h, but recovery of attraction took over 4 h, suggesting that habituation is also partially involved in reduced behavioural responsiveness following pre‐exposure. Seventy‐five min exposures to codlemone in moving air (5–10 cm s^?1) at rates of 0.9, 4.5, and 18 µg h^?1 from Celcon fibres caused 75, 86, and 99% disruption, respectively. However, 30–34‐h exposure of caged moths to air moving through an orchard treated with 1000 Isomate‐C® dispensers ha^?1 releasing approximately 20 µg h^?1 per dispenser during tests, had no impact on moth response in flight‐tunnel assays 30 min after removal from the orchard. In this treated orchard, catches of free‐flying moths in pheromone‐baited traps were completely inhibited. If observed mechanisms such as long‐lasting antennal adaptation or habituation of the central nervous system contribute to the disruption of pheromone communication among codling moths under field conditions, it seems unlikely that they occur following exposure to the average atmospheric concentrations of codlemone. For these effects to be important, codling moths may require close contact with pheromone sources for extended periods, or repeated close encounters.     

Pre-exposure modulates attraction to sex pheromone in a moth

In behavioural experiments we investigated the influence of previous short exposure to sex pheromone on subsequent response of male Spodoptera littoralis moths to sex pheromone. We found that pre-exposed males showed increased sensitivity to female sex pheromone after a single exposure to a pheromone plume compared to that found in na?ve males. The increased responsiveness lasted for at least 27 h after the exposure, showing that it was not just a short-term sensitization of the males. Exposure to the odour source without upwind movement towards the source was enough to increase the responsiveness. Physical activation without exposure to odour did not affect responsiveness. The increase in responsiveness after exposure was higher when the males were pre-exposed to natural female pheromone gland extract than when they were exposed to a higher dose of the main component, even though both odour sources elicited similar upwind attraction in na?ve males. Thus, the quality of the pheromone mixture to which males were exposed influenced the subsequent response.     

Differences between single cell responses to different components of the sex pheromone in males of the lightbrown apple moth (Epiphyas postvittana)

ABSTRACT. Single cell responses, elicited by each of three components of the pheromone blend of the lightbrown apple moth, Epiphyas postvittana (Walker) (Lepidoptera, Tortricidae), exhibit significant differences in disadaptation rates and effect of stimulation on the spontaneous generation of spikes upon removal of the stimulus. The major component has a disadaptation rate of a few seconds and a sustained effect on the rate of spontaneous spike generation upon removal of the stimulus. Faster disadaptation rates and sharp reduction in the spike rate upon removal of stimulus, were observed for one minor component, 12: Ac, and suggests a likely role in close-range orientation to the female. The second minor component produces two types of response, one of which resembles the major component. The other, more common response, exhibits a very slow rate of disadaptation, of the order of minutes. This characteristic also may have some bearing on search strategies.     

Representation of binary pheromone blends by glomerulus-specific olfactory projection neurons

An outstanding challenge in olfactory neurobiology is to explain how glomerular networks encode information about stimulus mixtures, which are typical of natural olfactory stimuli. In the moth Manduca sexta, a species-specific blend of two sex-pheromone components is required for reproductive signaling. Each component stimulates a different population of olfactory receptor cells that in turn target two identified glomeruli in the macroglomerular complex of the males antennal lobe. Using intracellular recording and staining, we examined how responses of projection neurons innervating these glomeruli are modulated by changes in the level and ratio of the two essential components in stimulus blends. Compared to projection neurons specific for one component, projection neurons that integrated information about the blend (received excitatory input from one component and inhibitory input from the other) showed enhanced ability to track a train of stimulus pulses. The precision of stimulus-pulse tracking was furthermore optimized at a synthetic blend ratio that mimics the physiological response to an extract of the females pheromone gland. Optimal responsiveness of a projection neuron to repetitive stimulus pulses therefore appears to depend not only on stimulus intensity but also on the relative strength of the two opposing synaptic inputs that are integrated by macroglomerular complex projection neurons.     

Circadian rhythmicity and development of the behavioural response to sex pheromone in male brown-banded cockroaches, Supella longipalpa

Abstract The effects of age, time of day and pheromone concentration on the responsiveness of the male brown-banded cockroach, Supella longipalpa (F.), to female volatile sex pheromone were examined. Male response increased with age and peaked 10 days after the imaginal moult. A diel periodicity of responsiveness was observed, with greater responses in the scotophase. The behavioural response was dose-dependent and its diel periodicity was most obvious when intermediate pheromone concentrations (e.g. 0.01 female equivalents) were used. Higher concentrations obscured the periodicity by eliciting greater responses in the photophase, while the lower concentrations did so by decreasing the response in the scotophase. Following entrainment to a LD 12:12 h cycle, the periodicity of response to 0.01 female equivalents of pheromone persisted for at least 54 h under continuous darkness, further demonstrating a true circadian rhythm.     

Binocular contributions to the responsiveness and integrative capacity of the circadian rhythm system to light

The retinohypothalamic tract (RHT), a monosynaptic retinal projection to the SCN, is the major path by which light entrains the circadian system to the external photoperiod. The circadian system of rodents effectively integrates or counts photons, and the magnitude of the rhythm phase response is proportional to the total energy of the photic stimulus. In the present studies, responsiveness to light and integrative capacity of the circadian system were tested in hamsters after reduction of retinal photoreceptor input by 50%. At CT 19, animals in constant darkness with or without unilateral retinal occlusion were exposed to 1 of 6 irradiances of 5-min white-light pulses ranging from 0.0011 to 70 microW/cm(2) or 5 white-light pulses of 0.6 microW/cm(2) with durations ranging from 0.25 to 150.0 min. Assessment of light-induced circadian rhythm phase response and Fos expression in the SCN by these animals revealed that a 50% reduction in input from photoreceptors stimulated directly with light caused a decrease in responsiveness to the longest duration and highest irradiance pulses presented. Despite this effect, both the magnitude of Fos induction in the SCN and phase-shift response remained directly proportional to the total energy in the photic stimuli. The results support the view that a reciprocal relationship between stimulus irradiance and duration persists despite the 50% reduction in retinal photoreceptor input. The mechanism of integration neither resides in the retina nor in the RHT.     

C75, a Fatty Acid Synthase Inhibitor,Inhibits Feeding Activity and Pheromone Production in a Moth,Helicoverpa zea

Fatty acid synthesis produces long-chain fatty acids that are principal forms of stored energy and essential constituents of cellular membrane lipids. In animals fatty acid synthesis is catalyzed by fatty acid synthase (FAS) from acetyl-coenyzyme A (CoA) and malonyl-CoA. Cerulenin and C75, potent FAS inhibitors, can inhibit feeding in mammals.Using these inhibitors we examined the effect of feeding inhibition during H. zea larval stage. Growth of larvae injected (30 μg/g body weight) with C75 or cerulenin was significantly delayed during the first 8 hrs after injection, but recovered to normal levels within 20 hrs. During the first 8 hr period, the amount of consumed diet in the inhibitor treated larvae was significantly less than the control group. The retardation of larval development could be caused from the reduction of food intake after injection of the inhibitor. The result indicates that C75 or cerulenin inhibits fatty acid synthesis, resulting in feeding suppression in the larval moth as demonstrated in vertebrates.Pheromone production was significantly decreased in the isolated pheromone gland of H. zea females treated with FAS inhibitors. Pheromone production was inhibited by blocking fatty acid synthesis, even though PBAN stimulated pheromone biosynthesis. After topical application of D3-16: Acid to pheromone glands the relative labeled pheromone amount was increased when the gland was incubated with C75. This result indicates that a part of the pheromone amount could be synthesized from 16: Acid directly when fatty acid synthesis was blocked. These results indicate that the inhibitors have a potential possibility to control insect feeding activity and inhibit pheromone biosynthesis in moths.     

No inhibitory effect on receptor neurone activity by sulphur analogues of the sex pheromone component (Z)-5-decenyl acetate in the turnip moth, Agrofis segetum (Lepidoptera: Noctuidae)

Abstract. Olfactory responses from the entire antenna and from single antennal sensilla of the male turnip moth, Agrotis segetum (Lepidoptea: Noctuidae Schiff.), were recorded after stimulation of the antenna with the sex pheromone component, (Z)-5-decenyl acetate (Z5-10:OAc), and three sulphur analogues of this compound. Adaptation of olfactory receptor neurones tuned to Z5-10:OAc was investigated after pre-exposure of these receptor neurones to the key stimulus and to the three sulphur analogues. Both electro-antennographic and single sensillum recordings revealed that the sulphur analogues had a significantly decreased effect compared to the natural stimulus. The pre-exposure experiments demonstrated that no further inhibition of neural activity was observed than could be expected from receptor neurone adaptation. Earlier reports, describing sulphur analogues as possible hyperagonists acting on moth pheromone receptor neurones, are not supported by the present study.     

Temperature dependent plasticity of habituation in the crayfish

To determine the effects of thermal preconditioning on a simple form of learning and memory, habituation, we preconditioned crayfish with extreme temperatures and subsequently analysed their effects on mechanosensory input that evokes a response in the lateral giant interneurons, within the normal temperature range of the animal. We found that repetitive stimulation with a 1 s interstimulus interval led to habituation of the response the lateral giant in control animals at 22°C. Neither heat nor cold preconditioning had any effect on the probability of evoking a response in the lateral giant nor on the rate at which habituation occurred. With a 1 min interstimulus interval, however, the rate of habituation of the lateral giant in the heat-preconditioned group was less than either the control or cold-preconditioned animals. The effect of heat or cold pre-exposure was specific to the input to the lateral giant at control temperatures. For example, at 22°C prior heat and cold preconditioning had no effect on spontaneous reductor motor neurone activity. They did, however, provide thermoprotection at extreme temperatures, with the probability of spontaneous activity higher in the cold-preconditioned group at low temperatures but higher in the heat-preconditioned group at higher temperatures.     

Increased EAG responses of tortricid moths after prolonged exposure to plant volatiles: evidence for octopamine-mediated sensitization

As measured by electroantennograms (EAG), both male and female obliquebanded leafrollers, Choristoneura rosaceana (Harris), and redbanded leafrollers, Argyrotaeniavelutinana (Walker), were similarly sensitive to host-related plant volatiles: trans-2-hexenal, benzaldehyde, 1-hexenol, cis-3-hexen-1-ol, geraniol, linalool, (+)-limonene, hexenal and trans-2-hexenol. Females of both species were similarly sensitive to the shared major component of their sex-attractant pheromone ((Z)11-14:Ac). Continuous 60 min pre-exposure of male and female C. rosaceana and A. velutinana to successively higher concentrations of a mixture of the nine plant volatiles in Teflon chambers with continuous air exchange caused a dosage-dependent increase in subsequent responsiveness (sensitization) to green leaf volatiles, as measured by EAGs. In addition, 60 min of pre-exposure of male C. rosaceana to certain individual volatiles ((+)-limonene, geraniol, benzaldehyde) increased EAGs nearly as much as did the mixture of nine volatiles. Pre-exposures to the nine plant-volatile mixture at concentrations achieved by 100 microg and 1 mg loading dosages in 100 microl of mineral oil significantly increased EAG depolarization to pheromone (cross-sensitization) in males but not females of both moth species. Antennae of male C. rosaceana pre-injected with 100 microg of octopamine (OA) without volatile pre-exposure exhibited sensitization nearly identical to that induced by pre-exposing moths to sensitizing concentrations of the plant-volatile mixture. Moreover, injection of the OA antagonist chlorpromazine (CP) blocked sensitization by the plant-volatile pre-exposure. Collectively, these findings suggest that exposures of tortricid moths to certain host-plant related volatiles may modulate subsequent olfactory sensitivity to behaviorally relevant chemical cues and that plant-volatile induced sensitization may be octopamine mediated.     

Habituation of the response to sex pheromone in two species of lepidoptera, with reference to a method of control

The rate of habituation of male Anagasta kuehniella to sources of sex pheromone at four different concentrations (10⁻⁴–10⁻¹ female equivalents of extract) was measured at the time of their diel peak of responsiveness. While the level of response to pheromone increased with concentration, the rate of habituation seemed to be independent of concentration provided this was above the threshold level for response. An interpretation of some data obtained with Trichoplusia ni by Shorey et al. yielded a similar conclusion: the rate of habituation of the response to pheromone was not greatly affected by changes in concentration. The relevance of these findings to the design of control measures for Lepidoptera is discussed.

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Zusammenfassung Die Geschwindigkeit, mit der sich m?nnliche Anagasta kuehniella an vier verschiedene Konzentrationen des Geschlechtspheromons (10⁻⁴–10⁻¹ Extrakt?quivalente weiblicher Tiere) gew?hnen, wurde w?hrend ihres t?glichen Empfindlichkeitsh?hepunktes gemessen. Das Reaktionsniveau stieg mit zunehmender Konzentration; die Gew?hnungsgeschwindigkeit schien, vorausgesetzt, da? die Reaktionsschwelle überhaupt überschritten wurde, jedoch unabh?ngig von der Konzentration zu sein. Die Auswertung der Resultate, die von Shorey et al. mit Trichoplusia ni erzielt wurden, lassen eine ?hnliche Schlussfolgerung zu: die Gew?hnungsgeschwindigkeit der Reaktion auf das Pheromon wurde durch Konzentrationsschwankungen nur wenig beeinflusst. Die Bedeutung der vorliegenden Resultate wird in Bezug auf Kontrollmassnahmen gegen Lepidopteren diskutiert.

     

Effect of pheromones,hormones, and handling on sucrose response thresholds of honey bees (Apis mellifera L.)

The responsiveness of bees to sucrose is an important indicator of honey bee foraging decisions. Correlated with sucrose responsiveness is forage choice behavior, age of first foraging, and conditioned learning response. Pheromones and hormones are significant components in social insect systems associated with the regulation of colony-level and individual foraging behavior. Bees were treated to different exposure regimes of queen and brood pheromones and their sucrose responsiveness measured. Bees reared with queen or brood pheromone were less responsive than controls. Our results suggest responsiveness to sucrose is a physiologically, neuronally mediated response. Orally administered octopamine significantly reduced sucrose response thresholds. Change in response to octopamine was on a time scale of minutes. The greatest separation between octopamine treated and control bees occurred 30 min after feeding. There was no significant sucrose response difference to doses ranging from 0.2 g to 20 g of octopamine. Topically applied methoprene significantly increased sucrose responsiveness. Handling method significantly affected sucrose responsiveness. Bees that were anesthetized by chilling or CO₂ treatment were significantly more responsive than control bees 30 min after handling. Sixty minutes after handling there were no significant treatment differences. We concluded that putative stress effects of handling were blocked by anesthetic.Abbreviations BP brood pheromone - JH juvenile hormone - OA octopamine - PER proboscis extension response - PER-RT proboscis extension response threshold - QMP queen mandibular pheromone     

L9 modulation of gill withdrawal reflex habituation in Aplysia.

Repeated tactile stimulation of the siphon in Aphysia normally results in habituation of the gill withdrawal reflex and a concomitant decrease in the amplitude of the excitatory synaptic input ot 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.