Responses of bombykol receptor cells to (Z,E)-4,6-hexadecadiene and linalool

Summary The bombykol analogue (Z,E)-4,6-hexadecadiene elicits nerve-impulse firing in the bombykol receptor cell of male moths ofBombyx mori. It is about 100 times less effective than bombykol if one compares equipotent loads of the odor sources. However, brief (1 s) hexadecadiene stimuli are followed by after-responses lasting for more than 10 min, much longer than those following bombykol stimuli yielding equal initial responses. The possibility is discussed that the unusual type of response to hexadecadiene is produced by a combination of long-lasting excitatory and shortlasting inhibitory effects on the receptor cell. Alternatively, hexadecadiene could be a weak activator which is converted on the antenna into a more potent compound. The after-responses can be reversibly inhibited by linalool stimuli. The electrophysiological effects observed led to the prediction that a single stimulus with (Z,E)-4,6-hexadecadiene followed by repetitive pulses of linalool should simulate a response to pulsed bombykol stimuli and elicit anemotactic behavior of maleBombyx mori.     

Termination of sex pheromone production in mated females of the silkworm moth

A mating duration of more than 6 h was necessary to permanently terminate the production of the sex pheromone (bombykol) in the silkworm moth, Bombyx mori L. (Lepidoptera: Bombycidae), although the female formed a bursa copulatrix including a spermatophore and laid fertilized eggs even after mating for only 0.5 h. The 6-h mated female again produced bombykol if given an injection of synthetic pheromonotropic neuropeptide (PBAN), which is known to activate pheromone biosynthesis in a virgin female. Extracts of brain-suboesophageal ganglion (SG) complexes, which were removed from 6- and 24-h mated females, showed strong pheromonotropic activities. These results indicated that the pheromone gland of the mated female maintained its ability to biosynthesize bombykol; however, it could not produce pheromone due to a suppression of PBAN secretion from the SG. Furthermore, bombykol titers did not decrease after mating in females with a transected ventral nerve cord, even after the injection of a spermatophore extract, suggesting that the suppression of PBAN secretion was mediated by a neural signal and not by a substance in the spermatophore. The mated females accumulated (10E, 12Z)-10,12-hexadecadienoic acid, a precursor of bombykol biosynthesis, in their pheromone glands as did decapitated females. © 1996 Wiley-Liss, Inc.     

A general odorant background affects the coding of pheromone stimulus intermittency in specialist olfactory receptor neurones

In nature the aerial trace of pheromone used by male moths to find a female appears as a train of discontinuous pulses separated by gaps among a complex odorant background constituted of plant volatiles. We investigated the effect of such background odor on behavior and coding of temporal parameters of pheromone pulse trains in the pheromone olfactory receptor neurons of Spodoptera littoralis. Effects of linalool background were tested by measuring walking behavior towards a source of pheromone. While velocity and orientation index did drop when linalool was turned on, both parameters recovered back to pre-background values after 40 s with linalool still present. Photo-ionization detector was used to characterize pulse delivery by our stimulator. The photo-ionization detector signal reached 71% of maximum amplitude at 50 ms pulses and followed the stimulus period at repetition rates up to 10 pulses/s. However, at high pulse rates the concentration of the odorant did not return to base level during inter-pulse intervals. Linalool decreased the intensity and shortened the response of receptor neurons to pulses. High contrast (>10 dB) in firing rate between pulses and inter-pulse intervals was observed for 1 and 4 pulses/s, both with and without background. Significantly more neurons followed the 4 pulses/s pattern when delivered over linalool; at the same time the information content was preserved almost to the control values. Rapid recovery of behavior shows that change of perceived intensity is more important than absolute stimulus intensity. While decreasing the response intensity, background odor preserved the temporal parameters of the specific signal.     

Use of bilateral information to determine the walking direction during orientation to a pheromone source in the silkmoth Bombyx mori

Odor source localization is an important animal behavior. Male moths locate mates by tracking sex pheromone emitted by conspecific females. During this type of behavior, males exhibit a combination of upwind surge and zigzagging flight. Similarly, the male walking moth Bombyx mori responds to transient pheromone exposure with a surge in movement, followed by sustained zigzagging walking. The initial surge direction is known to be influenced by the pheromone input pattern. Here, we identified the sensory input patterns that determine the initial walking direction of males. We first quantified the stimulus by measuring electroantennogram values, which were used as a reference for subsequent tests. We used a brief stimulus pulse to examine the relationship between sensory stimulus patterns and the turning direction of initial surge. We found that the difference in input timing and intensity between left and right antennae affected the walking direction, indicating that B. mori integrate bilateral pheromone information during orientation behavior. When we tested pheromone stimulation for longer periods, turning behavior was suppressed, which was induced by stimulus cessation. This study contributes toward understanding efficient strategies for odor-source localization that is utilized by walking insects.     

Representation of a mixture of pheromone and host plant odor by antennal lobe projection neurons of the silkmoth Bombyx mori

Pheromone-source orientation behavior can be modified by coexisting plant volatiles. Some host plant volatiles enhance the pheromonal responses of olfactory receptor neurons and increase the sensitivity of orientation behavior in the Lepidoptera species. Although many electrophysiological studies have focused on the pheromonal response of olfactory interneurons, the response to the mixture of pheromone and plant odor is not yet known. Using the silkmoth, Bombyx mori, we investigated the physiology of interneurons in the antennal lobe (AL), the primary olfactory center in the insect brain, in response to a mixture of the primary pheromone component bombykol and cis-3-hexen-1-ol, a mulberry leaf volatile. Application of the mixture enhanced the pheromonal responses of projection neurons innervating the macroglomerular complex in the AL. In contrast, the mixture of pheromone and cis-3-hexen-1-ol had little influence on the responses of projection neurons innervating the ordinary glomeruli whereas other plant odors dynamically modified the response. Together this suggests moths can process plant odor information under conditions of simultaneous exposure to sex pheromone.     

Inhibitory Contribution to Suprathreshold Corticostriatal Responses: An Experimental and Modeling Study

Neostriatal neurons may undergo events of spontaneous synchronization as those observed in recurrent networks of excitatory neurons, even when cortical afferents are transected. It is necessary to explain these events because the neostriatum is a recurrent network of inhibitory neurons. Synchronization of neuronal activity may be caused by plateau-like depolarizations. Plateau-like orthodromic depolarizations that resemble up-states in medium spiny neostriatal neurons (MSNs) may be induced by a single corticostriatal suprathreshold stimulus. Slow synaptic depolarizations may last hundreds of milliseconds, decay slower than the monosynaptic glutamatergic synaptic potentials that induce them, and sustain repetitive firing. Because inhibitory inputs impinging onto MSNs have a reversal potential above the resting membrane potential but below the threshold for firing, they conform a type of “shunting inhibition”. This work asks if shunting GABAergic inputs onto MSNs arrive asynchronously enough as to help in sustaining the plateau-like corticostriatal response after a single cortical stimulus. This may help to begin explaining autonomous processing in the striatal micro-circuitry in the presence of a tonic excitatory drive and independently of spatio-temporally organized inputs. It is shown here that besides synaptic currents from AMPA/KA- and NMDA-receptors, as well as L-type intrinsic Ca²⁺- currents, inhibitory synapses help in maintaining the slow depolarization, although they accomplish the role of depressing firing at the beginning of the response. We then used a NEURON model of spiny cells to show that inhibitory synapses arriving asynchronously on the dendrites can help to simulate a plateau potential similar to that observed experimentally. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Orientation of Aplysia californien to distant food sources

Summary The behavior of the marine mollusc Aplysia was examined under different experimental conditions designed to determine the food searching strategy of the animals. In a small, open field tank with still water, the animals took an average of 42 min to find a piece of seaweed, even though the stimulus was never located more than 30 cm away from the animal. Observations of the animals indicated that their search was not directed, without a clear tendency towards the food, and during the course of a search, they often crawled through most of the area of the tank. The search time, the distance travelled, and the strategy of the search of the animals was similar for different types of seaweed. If animals were aroused into activity by the presence of seaweed extract, the time for them to contact a piece of odorless glass fiber paper in the open field was not significantly different than that for a piece of seaweed. The probability at which the animals contacted the seaweed, as a function of the distance travelled, resembled the detection probability determined according to a theory of random search. We thus propose that the aroused animals move in a random pattern until they are very close to the food. This strategy can be advantageous in still water since chemicals do not provide distinct gradients that can serve as cues for chemotactic orientation from distances greater than a few centimeters from the source.In a Y-maze in still water, Aplysia did not perform above chance in selecting the arm that contained the seaweed. In streaming water, the animals entered the arm that contained seaweed significantly above chance only if a large piece of seaweed was used, so as to provide a very strong stimulus. Similarly, presentation of a high concentration of glutamate or a high concentration of seaweed extract to one side of the animal could evoke turning towards the stimulus. In the Y-maze, animals that repeatedly entered and exited both arms before making a choice, performed better than those that went directly into one arm.We conclude that chemical food stimuli can evoke directed orientation only when the animal is very close to the stimulus (within a few centimeters, at most). For Aplysia, the role of distant chemical stimuli may not be to guide their orientation, but rather to evoke the food-induced arousal state. In this behavioral state, several behaviors, such as locomotion are altered, and these changes may provide the basis for locating food from a distance.Abbreviation SWE seaweed extract     

Tracking the startle response of guppies Poecilia reticulata in three dimensions

A three‐dimensional analysis of startle behaviours of guppies Poecilia reticulata, in dyads or alone, from two populations that show distinct differences in shoaling behaviour was performed. During the first few seconds after a startling stimulus, changes in behaviour, which could be critical if an individual is to survive a predatory attack, and the interactions between pairs of P. reticulata were examined. The enhanced social interactions immediately after the stimulus, as a proxy for shoaling behaviour, and their dissipation were quantified. Social (individuals tested in dyads) v. asocial (tested alone) responses to the startling stimulus were also compared. The three‐dimensional reconstruction, from a two‐camera, high‐frame‐rate tracking system allowed for the tracking of the individuals' speed and speed recovery and, for P. reticulata in dyads, interindividual distance and orientation. For the dyads from the high‐predation population, the closer the individuals were to each other, the more likely they were to be parallel, but no correlation was found for the low‐predation P. reticulata. The startle response of P. reticulata comprised the following sequence: freezing, darting and skittering and recovery to pre‐stimulus swimming behaviour. Upon repeated encounters with the stimulus, a reduced shoaling and startle response was observed, although the rate of reduction was faster in P. reticulata from the high‐predation population than those from the low‐predation population. The results are discussed in light of what is known about the anti‐predator behaviour of this species.     

A single sex pheromone receptor determines chemical response specificity of sexual behavior in the silkmoth Bombyx mori

In insects and other animals, intraspecific communication between individuals of the opposite sex is mediated in part by chemical signals called sex pheromones. In most moth species, male moths rely heavily on species-specific sex pheromones emitted by female moths to identify and orient towards an appropriate mating partner among a large number of sympatric insect species. The silkmoth, Bombyx mori, utilizes the simplest possible pheromone system, in which a single pheromone component, (E, Z)-10,12-hexadecadienol (bombykol), is sufficient to elicit full sexual behavior. We have previously shown that the sex pheromone receptor BmOR1 mediates specific detection of bombykol in the antennae of male silkmoths. However, it is unclear whether the sex pheromone receptor is the minimally sufficient determination factor that triggers initiation of orientation behavior towards a potential mate. Using transgenic silkmoths expressing the sex pheromone receptor PxOR1 of the diamondback moth Plutella xylostella in BmOR1-expressing neurons, we show that the selectivity of the sex pheromone receptor determines the chemical response specificity of sexual behavior in the silkmoth. Bombykol receptor neurons expressing PxOR1 responded to its specific ligand, (Z)-11-hexadecenal (Z11-16:Ald), in a dose-dependent manner. Male moths expressing PxOR1 exhibited typical pheromone orientation behavior and copulation attempts in response to Z11-16:Ald and to females of P. xylostella. Transformation of the bombykol receptor neurons had no effect on their projections in the antennal lobe. These results indicate that activation of bombykol receptor neurons alone is sufficient to trigger full sexual behavior. Thus, a single gene defines behavioral selectivity in sex pheromone communication in the silkmoth. Our findings show that a single molecular determinant can not only function as a modulator of behavior but also as an all-or-nothing initiator of a complex species-specific behavioral sequence.     

Linalool odor stimulation improves heat stress tolerance and decreases fat accumulation in nematodes

Aromatherapy uses plant essential oils and fragrant ingredients for relaxation, sleep assistance, and improvement of restlessness related to dementia. Certain aromatic substances increase the life span and stress tolerance of nematodes. We investigated effects of exposure to linalool, a linear chain monoterpenic alcohol that is present in the essential oils of many plants, and its optical isomer, l-linalool, in Caenorhabditis elegans. Nematodes were repelled by the odor of both linalool and l-linalool; however, linalool odor stimulation decreased fat accumulation and increased motility after thermal stress. Analysis of a gene-deficient mutant revealed that the DAF-16 insulin-signaling pathway, which is involved in heat stress tolerance, was enhanced by linalool treatment. Linalool stimulation increased the expression of downstream genes such as sod-3 and hsp-12.6 via DAF-16. We conclude that linalool odor induces a repelling behavior in nematodes, improves heat stress tolerance through the DAF-16 signaling pathway, and affects fat accumulation.     

The Temporal Winner-Take-All Readout

How can the central nervous system make accurate decisions about external stimuli at short times on the basis of the noisy responses of nerve cell populations? It has been suggested that spike time latency is the source of fast decisions. Here, we propose a simple and fast readout mechanism, the temporal Winner-Take-All (tWTA), and undertake a study of its accuracy. The tWTA is studied in the framework of a statistical model for the dynamic response of a nerve cell population to an external stimulus. Each cell is characterized by a preferred stimulus, a unique value of the external stimulus for which it responds fastest. The tWTA estimate for the stimulus is the preferred stimulus of the cell that fired the first spike in the entire population. We then pose the questions: How accurate is the tWTA readout? What are the parameters that govern this accuracy? What are the effects of noise correlations and baseline firing? We find that tWTA sensitivity to the stimulus grows algebraically fast with the number of cells in the population, N, in contrast to the logarithmic slow scaling of the conventional rate-WTA sensitivity with N. Noise correlations in first-spike times of different cells can limit the accuracy of the tWTA readout, even in the limit of large N, similar to the effect that has been observed in population coding theory. We show that baseline firing also has a detrimental effect on tWTA accuracy. We suggest a generalization of the tWTA, the n-tWTA, which estimates the stimulus by the identity of the group of cells firing the first n spikes and show how this simple generalization can overcome the detrimental effect of baseline firing. Thus, the tWTA can provide fast and accurate responses discriminating between a small number of alternatives. High accuracy in estimation of a continuous stimulus can be obtained using the n-tWTA.     

Wind spectra and the response of the cercal system in the cockroach

Experiments on the cercal wind-sensing system of the American cockroach, Periplaneta americana, showed that the firing rate of the interneurons coding wind information depends on the bandwidth of random noise wind stimuli. The firing rate was shown to increase with decreases in the stimulus bandwidth, and be independent of changes in the total power of the stimulus with constant spectral composition. A detailed analysis of ethologically relevant stimulus parameters is presented. A phenomenological model of these relationships and their relevance to wind-mediated cockroach behavior is proposed.Abbreviations 2D two dimensional - FOWD fiber-optic wind detector - GI giant interneurons - STA spike-triggered average     

Dynamics of Encoding in a Population of Neurons

A simple encoder model, which is a reasonable idealization from known electrophysiological properties, yields a population in which the variation of the firing rate with time is a perfect replica of the shape of the input stimulus. A population of noise-free encoders which depart even slightly from the simple model yield a very much degraded copy of the input stimulus. The presence of noise improves the performance of such a population. The firing rate of a population of neurons is related to the firing rate of a single member in a subtle way.     

Bumble Bees Show Asymmetrical Discrimination Between Two Odors in a Classical Conditioning Procedure

Olfactory processing of two odorants and their mixture was investigated in bumble bees Bombus terrestris using classical conditioning of the proboscis extension. In a standard procedure, workers were able to learn linalool, phenylacetaldehyde, and the mixture of these two components, with a similar level of response to these three stimuli. Thereafter, when we applied a differential conditioning procedure where one rewarded odorant was presented alternately against an unrewarded one, an asymmetrical discrimination between the two pure odors was found. Bumble bees performed well in the discriminative task when linalool was the rewarded stimulus and phenylacetaldehyde the unrewarded one, but they had difficulty learning phenylacetaldehyde if it was the rewarded odor in the symmetrical procedure. Indeed, unrewarded stimulations with linalool appeared to disrupt the learning of the alternative odor, possibly due to an innate biological meaning of linalool.     

Interneurons sensitive to female pheromone in the deutocerebrum of the male silkworm moth, Bombyx mori

ABSTRACT. In response to minute quantities of female sex pheromone, the male silkworm moth, Bombyx mori L., walks upwind to locate the odour source. The axons of antennal receptors specific for the two known components of the pheromone terminate in the deutocerebrum. In this study, single interneurons were recorded extracellularly in the deutocerebrum of the male silkworm moth. Responses were characterized as the antennae were presented with puffs of clean air, or air containing either or both components of the female pheromone, bombykol and bombykal. An apparatus is described which added bombykol or bombykal to a constant air stream flowing over the antenna. Most units (87%) showed qualitatively different responses to bombykol and bombykal. A majority of the pheromone-sensitive units (65%) also showed mechanosensory responses to air puffs. Two units were recorded which were slightly inhibited by either bombykol or bombykal alone, but were excited by a mixture of the two.     

Evidence of self-sustained motoneuron firing in young and older adults.

Motoneurons demonstrate a type of self-sustained firing behavior that seems to be produced by a prolonged period of depolarization caused by intrinsic long-term changes in the motoneuron. Such self-sustained firing behavior has previously been reported in human motor units. The purpose of the present study was to investigate the occurrence of self-sustained firing behavior in older adults. Eight young (mean age 24 yrs) and eight older (mean age 73 yrs) individuals participated in the investigation. While subjects produced light dorsiflexion contractions, a brief vibration stimulus was applied to the tibialis anterior muscle. Motor unit recordings were also obtained from the tibialis anterior muscle. Self-sustained firing behavior was evidenced by the appearance of new motor unit recruitment following vibration, even as the motor units that fired before the vibratory stimulus maintained a steady firing rate. The proportion of motor units exhibiting self-sustained firing activity was similar in both young and older adults (approx. 23% of trials). We conclude that self-sustained firing behavior is a ubiquitous phenomenon that does not seem to be affected by the aging process.     

Deletion of the Bombyx mori odorant receptor co-receptor (BmOrco) impairs olfactory sensitivity in silkworms

Olfaction plays an essential role in many important insect behaviors such as feeding and reproduction. To detect olfactory stimuli, an odorant receptor co-receptor (Orco) is required. In this study, we deleted the Orco gene in the Lepidopteran model insect, Bombyx mori, using a binary transgene-based clustered regulatory interspaced short palindromic repeats (CRISPR)/Cas9 system. We initially generated somatic mutations in two targeted sites, from which we obtained homozygous mutants with deletion of a 866 base pair sequence. Because of the flight inability of B. mori, we developed a novel method to examine the adult mating behavior. Considering the specialization in larval feeding, we examined food selection behavior in Orco somatic mutants by the walking trail analysis of silkworm position over time. Single sensillum recordings indicated that the antenna of the homozygous mutant was unable to respond to either of the two sex pheromones, bombykol or bombykal. An adult mating behavior assay revealed that the Orco mutant displayed a significantly impaired mating selection behavior in response to natural pheromone released by a wild-type female moth as well as an 11:1 mixture of bombykol/bombykal. The mutants also exhibited a decreased response to bombykol and, similar to wild-type moths, they displayed no response to bombykal. A larval feeding behavior assay revealed that the Orco mutant displayed defective selection for mulberry leaves and different concentrations of the volatile compound cis-jasmone found in mulberry leaves. Deletion of BmOrco severely disrupts the olfactory system, suggesting that BmOrco is indispensable in the olfactory pathway. The approach used for generating somatic and homozygous mutations also highlights a novel method for mutagenesis. This study on BmOrco function provides insights into the insect olfactory system and also provides a paradigm for agroforestry pest control.     

Flight tunnel response of codling moth Cydia pomonella to blends of codlemone,codlemone antagonists and pear ester

Upwind orientation flights of codling moth males Cydia pomonella L. to a single source of sex pheromone (E,E)‐8,10‐dodecadienol (codlemone) are significantly reduced when blending it with pheromone antagonists, either with codlemone acetate, (E,E)‐8,10‐dodecadienyl acetate, or with the codlemone isomer (E,Z)‐8,10‐dodecadienol. However, once activated by a pheromone stimulus, males no longer distinguish between a pheromone source and these antagonistic blend sources. This shows that the pheromone stimulus required for the initiation of an upwind flight response differs from the stimulus for maintaining upwind flight and landing at the source. In contrast to pheromone antagonists, males discriminate between pheromone alone and a blend source of pheromone and the plant volatile pear ester, ethyl (2E,4Z)‐2,4‐decadienoate. This indicates a difference in the detection and neural integration of pheromone and plant volatile stimuli.     

Die Orientierung der Trichterspinne nach polarisiertem Licht

Summary The spider Agelena labyrinthica (Clerck) orients its movements by means of polarized light even if the reflection pattern of the web is not visible. If the spider is not permitted to see the overhead source of polarized light while the reflection pattern from the substratum is still visible, its orientation is not influenced by rotation of the polaroid. For this reason Agelena's orientation by means of polarized light must be based on a direct perception of the light's plane of vibration.     

Twitching and quivering of the tentacles during snail olfactory orientation

In Helix aspersa the posterior tentacles house a sensitive olfactory organ. We studied two types of tentacular movements, twitch and quiver. A twitch is a brief retraction (mean duration, 4.1 s); a quiver is a rapid lateral movement (350 ms) unaccompanied by retraction. We videotaped the tentacles while snails explored an open field. When an attractive odor source, linalool, was present at one side of the arena, the snails consistently moved towards it. By contrast, if only the carrier substance was present the snails moved in random directions. Twitching was 50 times more frequent during linalool trials than during control trials, while quivering was 1.4 times more frequent. Twitching increased steadily and dramatically as snails approached the linalool source and, in the temporal dimension, the maximum rate of twitching occurred when the snails arrived at the odor source. Quivers occurred at a fairly constant rate. Twitching is interpreted as a mechanism to remove odor molecules trapped in the liquid covering of the olfactory epithelium, thus resulting in better temporal resolution for olfactory perception. Quivering may be a mechanism to increase access of odor molecules to receptors by decreasing the boundary layer at the surface of the tentacle. Accepted: 24 May 1997