Electroantennogram Amplitudes Research Articles

Detection of Volatile Organic Compounds by Antennal Lamellae of a Scarab Beetle

Chemoreceptive sensilla are abundantly distributed on antennal lamellae of scarab beetles. Olfactory reception by these sensory lamellae plays a major role in feeding behaviors and sexual communication of these beetles. A new electroantennogram (EAG) recording technique is here described for evaluation of electrophysiological responses of antennal lamellae of Pseudosymmachia flavescens to sex pheromones and host plant-related compounds. EAG responses were recorded simultaneously from each lamella and the closed antennal club. All test stimuli elicited similar EAG depolarization profiles in all the three lamellae and the closed club although EAG amplitudes from the same lamella or the club varied widely among different chemical stimuli. The mid lamella tended to produce significantly greater EAG responses. EAG responses evoked by a sex pheromone component, anisole, showed a significant correlation with the density of sensilla placodea subtype 1 (SP1). However, no general patterns were obtained for correlations between the density of any test sensilla type and EAG amplitudes evoked by all the six plant volatiles. Single sensillum recordings are needed to elucidate the specific roles of these sensilla in intraspecific sexual communication and perception of host plant volatiles.

Real-Time Odor Discrimination Using Single Antenna of Insect

In this letter, a system that discriminates between two odorants in real-time with a single antenna of an insect was constructed. In previous studies, odorants were discriminated through arraying multiple types of sensors. However, the use of a sensor array enlarges the entire system and requires complex signal processing, which makes it difficult to mount the system on a quadcopter with limited payloads or an autonomous robot with low computational power. On the other hand, the antenna of an insect is composed of several olfactory receptors; thus, a single antenna responds to multiple odorants. Therefore, a system for discriminating between multiple odorants from the electroantennogram (EAG) signal of a single antenna of an insect is proposed. An antenna of an adult male silkmoth is employed; the silkmoth antenna exhibits an electric potential change when detecting an odor, but the difference in the EAG amplitude and recovery time depends on the type of odorant. Since EAG is a type of neural signal, it is difficult to perform odor discrimination by simply setting the threshold, because it drifts due to noise. Therefore, the Hodgkin-Huxley model (HH model) was applied to the raw EAG signal as a dynamic filter, and the spike firing rate of the output value was calculated using the HH model. Then, the odorants were discriminated against based on the magnitude of the spike firing rate. A series of signal processing was implemented in a microcontroller, and the experiments indicated that all antennae demonstrated a discrimination performance of 90$\%$ or more.

Association between olfactory sensitivity and behavioral responses of Drosophila suzukii to naturally occurring volatile compounds.

Drosophila suzukii Matsumura (Diptera: Drosophilidae) is an invasive, destructive crop pest that originated in South East Asia. D. suzukii recently invaded Western countries and is threatening both European and American fruit industries. It is extremely attracted to otherwise undamaged, ripening fruits, unlike most other Drosophila species that attack only decaying or rotten fruits. Recent studies on different insect species showed that several naturally occurring compounds of easy market availability showing deterrent action may be used to supplement mass catches with food traps. Based on these considerations, the aim of the present work was to test the effects of some natural compounds (alone or in the mixture) on the olfactory system of the D. suzukii and the behavioral responses evoked. We measured by electroantennogram (EAG) recordings, the olfactory sensitivity of antennae to increasing concentrations of eugenol, vanillin, menthol, cis-jasmone; eugenol + vanillin, +menthol, +cis-jasmone; vanillin + menthol, +cis-jasmone. In addition, the behavioral responses to the same compounds and mixtures were evaluated. Our electrophysiological results show a dose-response relationship between the EAG amplitudes and the increasing concentrations of the olfactory compound. The behavioral results show that the number of laid eggs is significantly different between the standard diet and the standard diet + natural compound. These results underline a specificity in the olfactory sensitivity and in the ovipositing behavior of D. suzukii females; also, they could be valuable for the identification of key chemicals aimed at the future development of strategies in the management and control of this harmful insect for crops.

Olfactory sensitivity to major, intermediate and trace components of sex pheromone in Ceratitis capitata is related to mating and circadian rhythm

The Mediterranean fruit fly, Ceratitis capitata Wied., is a worldwide pest of several fruits given its extremely wide host range which includes more than 250 different species of fruits and vegetables. Its high biological potential is mainly due both to its ability to readily adapt to new environments and its high reproductive capacity as it completes multiple generations each year. Since sexually mature males emit a sex pheromone to call both other males for “lekking” and receptive females for mating, many studies have been directed to characterize the chemical composition of the sex pheromone. Besides, the release of sex pheromone appears to be modulated both by mating and time of day. Based on these considerations, we measured the olfactory sensitivity of antennae and palps of C. capitata to six volatiles of the male sex-pheromone: α-farnesene and geranyl acetate (major components), linalool and β-myrcene (intermediate components), β-farnesene and 2,3-butanediol (minor/trace components). The electroantennogram (EAG) and electropalpogram (EPG) responses were evaluated in both sexes, at different physiological states (virgin and mated), and at different times of the day (morning and afternoon). The results show that the EAG amplitude values in response to all stimuli are higher in the morning than in the afternoon for both sexes and in both virgin and mated insects. Furthermore, in both sexes, the olfactory sensitivity of virgin insects is higher than in mated ones. The EPG amplitude in response to all stimuli is higher in the morning in mated females than in virgin females and higher in the morning than in the afternoon in both mated sexes. By gaining knowledge on the effects of sex, physiological state and time of day on the olfactory sensitivity of C. capitata, one could better understand the medfly reproductive behavior.

Electroantennographic response of rice moth Corcyra cephalonica stainton to some plant volatiles

Electro antennogram (EAG) responses from male and female antennae of rice moth Corcyra cephalonica (Lepidoptera: Pyralidae) were observed for 18 plant volatiles at four concentrations (0.1, 1.0, 10 and 100 mg/ml). Mean EAG amplitude relative to hexanal, which was used as standard, revealed the significant differences in response to these from both male and female antennae. Among all the compounds (R)-(+)-Limonene, hexanal, cis-jasmone and acetophenone were found to be eliciting significant response from both male and female antennae.

Differential Electrophysiological Responses to Odorant Isotopologues in Drosophilid Antennae.

Olfaction presents the ultimate challenge to molecular recognition as thousands of molecules have to be recognized by far fewer olfactory receptors. We have presented evidence that Drosophila readily distinguish odorants based on their molecular vibrations using a battery of behavioral assays suggesting engagement of a molecular vibration-sensing component. Here we interrogate electrophysiologically the antennae of four Drosophilids and demonstrate conserved differential response amplitudes to aldehydes, alcohols, ketones, nitriles, and their deuterated isotopologues. Certain deuterated odorants evoked larger electroantennogram (EAG) amplitudes, while the response to the normal odorant was elevated in others. Significantly, benzonitrile isotopologues were not distinguishable as predicted. This suggests that isotopologue-specific EAG amplitudes result from differential activation of specific olfactory receptors. In support of this, odorants with as few as two deuteria evoke distinct EAG amplitudes from their normal isotopologues, and this is independent of the size of the deuterated molecule. Importantly, we find no evidence that these isotopologue-specific amplitudes depend on perireceptor mechanisms or other pertinent physical property of the deuterated odorants. Rather, our results strongly suggest that Drosophilid olfactory receptors are activated by molecular vibrations differentiating similarly sized and shaped odorants in vivo, yielding sufficient differential information to drive behavioral choices.

Conophthorin enhances the electroantennogram and field behavioural response ofXylosandrus germanus(Coleoptera: Curculionidae) to ethanol

Ethanol acts as an attractant that aids Xylosandrus germanus (Blandford) in locating living but weakened hosts. Electroantennogram (EAG) and field trapping experiments were conducted with conophthorin and other selected semiochemicals that function as important olfactory cues for various ambrosia beetles to characterize their effect on the attraction of X. germanus to ethanol and to assess whether EAG responses provide an indication of behavioural activity. Thus, semiochemicals hypothesized to enhance (i.e. conophthorin), reduce (i.e. terpinolene, verbenone) or have a negligible (i.e. lineatin, sulcatol, retusol) effect on the attraction of X. germanus to ethanol were selected for comparative purposes. Ethanol plus conophthorin elicited larger EAG amplitudes than binary (1 : 1) mixtures of ethanol plus the remaining semiochemicals at dilutions of 0.001%, 0.01% and 1% per compound. Conophthorin also enhanced the attraction of X. germanus, Euwallacea validus (Eichhoff) and Cyclorhipidion pelliculosum (Eichhoff) to ethanol under field conditions. By contrast, terpinolene and verbenone reduced the attraction of X. germanus and other ambrosia beetles to ethanol. Semiochemicals that enhance (i.e. conophthorin) or reduce (i.e. terpinolene, verbenone) attraction by X. germanus may improve monitoring and/or management tactics. These results also suggest that EAG responses can indicate field behavioural activity and be useful for screening attractants for X. germanus.

Circadian Regulation of Olfactory Receptor Neurons in the Cockroach Antenna

In the cockroach, olfactory sensitivity as measured by the amplitude of the electroantennogram (EAG) is regulated by the circadian system. We wished to determine how this rhythm in antennal response was reflected in the activity of individual olfactory receptor neurons. The amplitude of the EAG and the activity of olfactory receptor neurons (ORNs) in single olfactory sensilla were recorded simultaneously for 3 to 5 days in constant darkness from an antenna of the cockroach Leucophaea maderae. Both EAG amplitude and the spike frequency of the ORNs exhibited circadian rhythms with peak amplitude/activity occurring in the subjective day. The phases of the rhythms were dependent on the phase of the prior light cycle and thus were entrainable by light. Ablation of the optic lobes abolished the rhythm in EAG amplitude as has been previously reported. In contrast, the rhythm in ORN response persisted following surgery. These results indicated that a circadian clock outside the optic lobes can regulate the responses of olfactory receptor neurons and further that this modulation of the ORN response is not dependent on the circadian rhythm in EAG amplitude.

Pinocchio, a novel protein expressed in the antenna, contributes to olfactory behavior inDrosophila melanogaster

Most organisms depend on chemoreception for survival and reproduction. In Drosophila melanogaster multigene families of chemosensory receptors and putative odorant binding proteins have been identified. Here, we introduce an additional distinct protein, encoded by the CG4710 gene, that contributes to olfactory behavior. Previously, we identified through P[lArB]-element mutagenesis a smell impaired (smi) mutant, smi21F, with odorant-specific defects in avoidance responses. Here, we show that the smi21F mutant also exhibits reduced attractant responses to some, but not all, of a select group of odorants. Furthermore, electroantennogram amplitudes are increased in smi21F flies. Characterization of flanking sequences of the P[lArB] insertion site, complementation mapping, phenotypic reversion through P-element excision, and expression analysis implicate a predicted gene, CG4710, as the candidate smi gene. CG4710 produces two transcripts that encode proteins that contain conserved cysteines and which are reduced in the smi21F mutant. Furthermore, in situ hybridization reveals CG4710 expression in the third antennal segment. We have named this gene of previously unknown function and its product "Pinocchio (Pino)".

Behavioral and electrophysiological responses of the brownbanded cockroach, Supella longipalpa, to stereoisomers of its sex pheromone, supellapyrone.

Females of the brownbanded cockroach, Supella longipalpa, release a sex pheromone (supellapyrone) during a calling behavior and attract males from a distance. Supellapyrone has four possible configurations resulting from two asymmetric carbons at positions 2 and 4 (i.e., 2R,4R; 2R,4S; 2S,4R; and 2S,4S), but only the RR isomer is produced by females. Using pure synthetic stereoisomers in field tests, we showed that males are attracted to RR but also to high concentrations of the isomer SR. To study the activity of the stereoisomers in more detail we developed behavioral and electroantennogram (EAG) dose-response curves for each. Behaviorally, RR was the most active isomer with just 0.3 pg delivered on a filter paper being sufficient to elicit 50% male response in the olfactometer. Males were also attracted to SR and SS in the olfactometer, but at much higher dosages (100x ) than the natural compound; RS did not elicit behavioral responses at any of the doses tested. In EAG assays, the antenna of male S. longipalpa showed high and similar sensitivity to RR and SR, but a much lower (10%) sensitivity to SS and practically no response to RS. The lack of agreement between behavioral and electrophysiological data suggested either that RR and SR stimulate different antennal sensory neuron types, or that some aspect of the interaction between the pheromone and the sensillum environment or the receptor neuron itself is different. To test the first hypothesis we examined the response of the antenna before and after adaptation with each of the four stereoisomers. Positive cross-adaptation between RR and SR suggests that these two compounds stimulate the same receptor cells. Therefore, the lack of agreement between behavioral and EAG dose-response curves could be explained by isomer-specific molecular interactions between the pheromone and the receptor neuron. Although RR and SR produced the same EAG amplitude, stimulation with SR resulted in a slower recovery rate (i.e., wider peaks) than stimulation with RR. To gain further understanding of the response specificity of the antennae to the different stereoisomers we compared EAG responses (amplitude and recovery time) in response to individual stereoisomers and binary mixtures of isomers. These tests showed additive responses of the EAG amplitude to mixtures of compounds, but nonadditive responses of EAG recovery time. Therefore, peak height and width are independent parameters of the EAG, probably representing different intrasensillar events, and likely resulting in the expression of different behavioral responses.

Improvement of signal-to-noise ratio in electroantennogram responses using multiple insect antennae

Using an array of insect antennae connected in series or in parallel, electroantennogram (EAG) responses and noise levels were investigated in an attempt to improve signal-to-noise (S/N) ratio and sensitivity. Both the EAG response amplitude and noise level increased when the antennae of male Helicoverpa zea (Lepidoptera: Noctuidae) were connected in series. Due to lower relative increase in noise level than EAG amplitude as the number of antennae increased, the S/N ratio was also significantly improved by the serial connection. As a result the sensitivity of EAG was improved by the serial connection, which showed ca. ten-fold improvement in the threshold detection levels compared with a single antenna when four antennae were connected in series. In contrast to the serial connection, there were no differences in EAG amplitudes and overall noise levels when different numbers of antennae were connected in parallel. When only large-amplitude noise was taken into account, however, the S/N ratio was somewhat improved by the parallel connection. The frequency of overall noise remained at the same level both in the serial and in the parallel connection. However, the frequency of the large-amplitude noise increased in serial connection but decreased in parallel connection. The present study clearly indicates that both the sensitivity and S/N ratio of the EAG biosensor could be significantly improved by using the multiple antennal connections.

Odor Discrimination using Insect Electroantennogram Responses from an Insect Antennal Array

Insects have a highly developed olfactory sensory system, mainly based in their antennae, for the detection and discrimination of volatile compounds in the environment. Electroantennogram (EAG) response profiles of five different insect species, Drosophila melanogaster, Heliothis virescens, Helicoverpa zea, Ostrinia nubilalis and Microplitis croceipes, showed different, species-specific EAG response spectra to 20 volatile compounds tested. The EAG response profiles were then reconstructed for each compound across the five insect species. Most of the compounds could be distinguished by comparing the response spectra. We then used a four-antenna array, called a Quadro-probe EAG, to see if we could discriminate among odorants based on the relative EAG amplitudes evoked when the probe was placed in plumes in a wind tunnel and in a field. Stable EAG responses could be simultaneously and independently recorded with four different insect antennae mounted on the Quadro-probe, and different volatile compounds could be distinguished in real time by comparing relative EAG responses with a combination of differently tuned insect antennae. Regardless of insect species or EAG amplitudes, antennae on the Quadro-probe maintained their responsiveness with higher than 1 peak/s of time resolution.

Acj6: a gene affecting olfactory physiology and behavior in Drosophila.

Mutations affecting olfactory behavior provide material for use in molecular studies of olfaction in Drosophila melanogaster. Using the electroantennogram (EAG), a measure of antennal physiology, we have found an adult antennal defect in the olfactory behavioral mutant abnormal chemosensory jump 6 (acj6). The acj6 EAG defect was mapped to a single locus and the same mutation was found to be responsible for both reduction in EAG amplitude and diminished behavioral response, as if reduced antennal responsiveness to odorant is responsible for abnormal chemosensory behavior in the mutant. acj6 larval olfactory behavior is also abnormal; the mutation seems to alter cellular processes necessary for olfaction at both developmental stages. The acj6 mutation exhibits specificity in that visual system function appears normal in larvae and adults. These experiments provide evidence that the acj6 gene encodes a product required for olfactory signal transduction.

The electroantennogram response: Effects of varying sensillum numbers and recording electrode position in a clubbed antenna

The antennal response of Oryzaephilus surinamensis (Coleoptera: Silvanidae) to a food odour (carob) was used to investigate the separate effects of sensillum number and electrode position on the electroantennogram recorded from a clubbed antenna. By successive removal of club segments, it was shown that the electroantennogram amplitude was proportional to the number of olfactory sensilla remaining on the antenna. Varying the recording electrode position revealed a linear gradient in the electroantennogram over the non-club segments (2–8), where no olfactory sensilla were present, but a uniform electroantennogram over the club segments (9–11) where olfactory sensilla were present. An electrical equivalent network is proposed for the antenna which explains these results, and the non-uniformity of techniques classified as electroantennograms is discussed.

Male european corn borer,Ostrinia nubilalis (H�bner), antennal responses to analogs of its sex pheromone

Experiments were conducted to (1) determine whether the electroantennogram (EAG) can detect differences among the responses of antennae from males derived from the three strains ofOstrinia nubilalis (Hübner), and (2) characterize the EAG responses of each strain to isomeric forms of the natural pheromone, (E)- and (Z)-11-tetradecen-1-ol acetate (TDA), and analogs possessing differences in the terminal alkyl group, cyclopropyl (CPA), ortert-butyl (TBA).EAG responses differed among the strains in two ways: (1) Antennae fromZZ males always produced an EAG to (Z)-TDA with an extended duration of response. This "signature" EAG response was found to be unique to the antennal response ofZZ males to (Z)-TDA, thus providing a relatively easy method of distinguishing liveZZ males fromEE orZE males. Correlated with this longer EAG response was a longer disadaptation time, i.e., the EAG response ofZZ antennae disadapted more slowly (ca. 10 min) than the response ofEE antennae. (2) Strain differences in the relative EAG amplitudes to isomers and analogs were observed at the stimulus amounts eliciting the peak EAG amplitude as follows: TDA ≥ CPA > TBA forZZ males and both isomers; TDA > CPA ≥ TBA and CPA ≥ TDA > TBA forEE males and theE andZ isomers, respectively; CPA > TBA ≥ TDA forZE males and both isomers. Dose-response relationships were seen for all compounds if amplitude ("peak height") of the EAG was used as a measure of response. However, if width of the EAG at half the peak height ("peak width") was used, then only theZZ antennal response to (Z)-TDA resulted in a meaningful dose-response relationship. For all strains, the EAG amplitudes elicited by theZ isomers of any of the tested compounds were greater than those elicited by the correspondingE isomers. Therefore, correlations between the relative EAG and upwind flight responses were observed in theZZ (r = 0.86) andZE (r = 0.80) strains but were not correlated in theEE strain (r = 0.18). Temporal studies showed that adaptation, not postexcision deterioration, was responsible for the observed decreases in the EAG amplitude after repetitive stimulation or after stimulation with amounts in a descending order. Disa-daptation required at least 20 min for a moderate dose (10 μg for 1 sec). Developmental studies showed that antennae from 2-day-old adults had the greatest EAG response.

Field and laboratory electroantennographic measurements of pheromone plume structure correlated with oriental fruit moth behaviour

Abstract. Peak‐to‐trough electroantennogram amplitudes (bursts), caused by the individual filaments of a plume of female pheromone, diminish as high‐emission‐rate sources are approached by male Grapholita molesta, and this reduction is correlated with in‐flight arrestment (ceasing to advance upwind). These findings are consistent with the hypothesis that one cause of in‐flight arrestment in response to high‐concentration point sources is the attenuation of the peak‐to‐ trough amplitudes close to the source. High burst frequency, high pheromone flux, or low levels of continuous neuronal activity all are less well correlated with arrestment. Rather, arrestment appears due to a reduction of chemosensory input to the CNS during flight up the plume, even though the actual molecular concentration continues to increase. In a laboratory wind tunnel, upwind flight initiation by more than 20% of males was elicited only by pheromone source concentrations evoking significant fluctuations in EAG amplitudes at downwind release points. The burst frequencies that evoked high levels of upwind flight initiation ranged from a mean of 0.4‐2.2 bursts/s. Because a previous study revealed that flying male G. molestachange their course angle within 0.15 s of losing or contacting pheromone, these EAG burst frequencies indicate that during flight in a pheromone plume, many manoeuvres are probably made in response to contact with individual plume filaments. Thus, upwind flight tracks may be shaped by hundreds of steering reactions in response to encounters with individual pheromone filaments and pockets of clean air. Field‐recorded EAGs reveal that burst amplitudes diminish from 3 to 30 m downwind of the source, whereas burst frequencies do not, averaging c.1/s at 3, 10 and 30 m downwind.

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.

Electroantennogram response gradient on the antenna of the European corn borer, Ostrinia nubilalis

The electroantennogram (EAG) response of male moths of the European corn borer ( Ostrinia nubilalis) to their two pheromone components, (Z)- and (E)-11-tetradecenyl acetates, were studied. The EAG amplitude of the fast initial phase was approximately proportional to the antennal length. Local EAG responses were obtained along the length of the antenna with multiple electrodes. Response-interaction studies indicated a unique spread of the EAG response in a proximal direction in the antenna. A possible mechanism of the EAG summation was considered. The EAG response characteristic was similar for both (Z)- and (E)-11-tetradecenyl acetate along the length of the antenna.