Temporal resolution of odour pulses by three types of pheromone receptor cells inAntheraea polyphemus

Summary Temporal response characteristics of three cell types of maleA. polyphemus, each responding to a different pheromone component, have been measured using series of short (20 ms) pheromone pulses. The stimuli were delivered through capillaries 20 m in diameter and applied to single olfactory sensilla trichodea. Two of three cell types sensitive to (E,Z)-6,11-hexadecadienal and (E,Z)-4,9-tetradecadienyl acetate are able to resolve at least 5 stimuli/s whereas the third, responding to the major pheromone component (E,Z)-6,11-hexadecadienyl acetate, is slower, resolving only about 2 stimuli/s. These results suggest that receptor cells are able to respond to pulses of pheromone concentration as they occur downwind from a point source. The time-averaged number of nerve impulses does not seem to be a reliable measure of the amount of pheromone reaching the sensillum. Responses of the cells thus reflect the non-uniform distribution of pheromone in a plume rather than the average concentration.     

Repetitive stimulation of olfactory receptor cells in female silkmoths Bombyx mori L

The pheromone-sensitive receptor cells of male moth antennae are capable of detecting the rapid changes in stimulus intensity encountered in natural pheromone odour plumes. We investigated temporal response characteristics of the two receptor cell types of the sensillum trichodeum of female Bombyx mori, which are most sensitive to benzoic acid and 2,6-dimethyl-5-heptene-2-ol (DMH), respectively. The cells were repetitively stimulated with 50-ms pulses of benzoic acid and (±)-linalool, an effective mimic of DMH, at various pulse rates and different stimulus intensities. By recording receptor potentials and nerve impulses we demonstrated that both receptor cell types were able to follow stimulus pulses at least up to eight pulses per sec, with a more pronounced modulation of the responses in the DMH cell. The resolution capability of the two cell types showed little dependence on stimulus intensity. In their ability to resolve pulsed odour stimuli, the receptor cells for benzoic acid and DMH were as good as pheromone receptor cells.     

Octopamine modulates the sensitivity of silkmoth pheromone receptor neurons

Effects of octopamine and its antagonist epinastine on electrophysiological responses of receptor neurons of Antheraea polyphemus specialised to the pheromone components (E,Z)-6,11-hexadecadienyl acetate and (E,Z)-6,11-hexadecadienal were investigated. Injections of octopamine and epinastine into the moths had no effect on the transepithelial potential of the antennal-branch preparation nor on the spontaneous nerve impulse frequency in either type of receptor neuron. However, in the presence of continuous low-intensity pheromone stimulation, octopamine significantly increased the nerve impulse frequency in the acetate receptor neuron, but not in the aldehyde receptor neuron. Octopamine and epinastine had no significant effect on the receptor potential amplitudes elicited in both receptor neuron types by pheromone stimulation. However, the peak nerve impulse frequency in the response of both receptor neuron types to pheromone was significantly affected: decreased by epinastine and increased by octopamine over a broad range of pheromone concentrations. In control experiments, injection of physiological saline did not significantly alter the peak nerve impulse frequency. The effect of octopamine was established within 1 h after injection and persisted for about 4 h. The possibility of a direct action of octopamine on the nerve impulse generation by the receptor neurons is discussed. Accepted: 8 January 2000     

Peripheral Mechanisms of Pheromone Reception in Moths

Moths pheromones mostly consist of two or a few chemical componentsin a species-specific ratio. Each component is perceived bya particular type of receptor cell. Some pheromone componentscan inhibit the behavioral responses to other pheromone components.A single pheromone molecule is sufficient to elicit a nerveimpulse. The dose-response curve of single pheromone receptorneurons increases over many decades of stimulus intensity. Pheromonereceptor cells can resolve single stimulus pulses up to a frequencyof 10 pulses/s. Electrophysiological and biochemical studieson perireceptor events suggest that the pheromone moleculesinteract with the receptor cell while bound to a reduced formof the pheromone binding protein. The enzymatic degradationof pheromone found on the antennae is much too slow to accountfor the decline of the receptor potential after end of stimulation.The postulated rapid deactivation of the odor molecules adsorbedmight be performed by an oxidation of the pheromone bindingprotein. Several second messenger systems seem to be involvedin the cellular transduction mechanism (IP₃ diacylglycerol,cGMP, Ca₂₊). It is, however, not excluded that pheromone moleculescan gate single ion channels directly and thus elicit the elementaryreceptor potentials, observed at weak stimulus intensities.Chem. Senses 21: 257–268, 1996.     

Fine-scale resolution of closely spaced pheromone and antagonist filaments by flying male Helicoverpa zea

The limits of a male moth's ability to resolve closely spaced odor filaments have been investigated. Male Helicoverpa zea normally respond to their conspecific sex pheromone blend by exhibiting an upwind flight, which culminates in source contact by at least 50% of the bioassayed individuals. When loaded onto the same filter paper source containing this hitherto attractive pheromone blend, or onto a separate filter paper and co-emitted from the same pipette source with pheromone, (Z)-11-hexadecenyl acetate severely reduced upwind flight and source contact by male H. zea. A similar level of upwind flight inhibition was recorded when the antagonist (Z)-11-hexadecenyl acetate was emitted from its own point source placed 1 mm upwind of the pheromone point source, both plumes being simultaneously emitted in a continuous mode to form a confluent strand. However, (Z)-11-hexadecenyl acetate was less effective in reducing upwind flight and source contact when it was isolated and pulsed from its own source, placed 1 mm either upwind, downwind or cross-wind of a pipette source from which pheromone was simultaneously being pulsed, such that both filaments were separated in time by 0.001–0. 003 s. These results suggest that male H. zea are able to distinguish between odor sources separated by as little as 1 mm in space and 0.001 s in time. Accepted: 31 March 1999     

Inhibitors of sensillar esterase reversibly block the responses of moth pheromone receptor cells

Three volatile alkyl-thio-trifluoro propanones inhibiting the esterase in olfactory sensilla of the silkmoths Antheraea polyphemus and A. pernyi were used to test the hypothesis that enzymatic pheromone degradation is responsible for the decline of the receptor potential after pheromone stimulation. Test stimuli were the pheromone components (E,Z)-6,11-hexadecadienyl acetate, a substrate for the sensillar esterase, and (E,Z)-6,11-hexadecadienal, not degraded by the esterase. Each compound acts on a separate type of receptor cell. In both receptor cell types the trifluoro propanones caused a partially reversible reduction of sensitivity as indicated by smaller receptor potential amplitudes and lower nerve impulse frequencies. Since application of the esterase inhibitors did not prolong the decline of the receptor potential of the acetate cell, the esterase is not responsible for the rapid pheromone deactivation. When the trifluoro propanones were applied after the pheromone at high concentrations, they rapidly inhibited (repolarized) both receptor cell types. Experiments with local application of trifluoro propanones revealed that the inhibitory effect spreads within seconds along the length of the sensillum. The inhibition of the electrophysiological responses might be due to an antagonistic action of the trifluoro propanones at the pheromone-binding sites, either at the receptor molecules or at the pheromone-binding protein. Accepted: 4 February 1998     

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.     

Antennal resolution of pulsed pheromone plumes in three moth species

Male antennae of Cadra cautella,Pectinophora gossypiella, and Spodoptera exigua were presented with 20-ms-duration pulses of their two-component pheromone at rates of 1 to 33 Hz. Fourier analyses of electroantennograms resolved the temporal structure of trains of pheromone filaments delivered at up to 33 Hz for C. cautella and S. exigua and 25 Hz for P. gossypiella. Pheromone components tested separately for each species were generally equivalent in filament resolution to complete blends. Ambient temperatures of 18, 23 and 28 °C affected filament resolution only slightly, with poorer ability to discriminate rapidly pulsed signals at 18 °C. The question of how, or indeed if, such frequencies are conserved beyond the peripheral nervous system, remains.     

Temporal coding of pheromone pulses and trains in Manduca sexta

Summary We investigated the ability of pheromone-sensitive olfactory receptors of male Manduca sexta to respond to 20-ms pulses of bombykal, the major component of the conspecific pheromonal blend. Isolated pulses of bombykal elicited a burst of activity which decreased exponentially with a time constant of 160–250 ms. Trains of pulses delivered at increasing frequencies (0.5–10 Hz) elicited temporally modulated responses at up to 3 Hz. Concentration of the stimulus (1, 10, 100 ng per odor source) had a marginal effect on the temporal resolution of the receptors. Within a train, the responses to individual pulses remained constant, except for 10-Hz trains (short-term adaptation). A dose-dependent decline of responsiveness was observed during experiments (long-term adaptation). Although individual neurons may not respond faithfully to each pulse of a train, the population of receptors sampled in this study appears to be capable of encoding the onset of odor pulses at frequencies of up to at least 3 Hz.Abbreviations BAL bombykal or (E,Z)-10,12-hexadecadienal - C15 (E,Z)-11,13-pentadecadienal - HAL (E)-2-hexenal - EAG electroantennogram - P1, P2, P3 single stimulus pulses - PSTH peri-stimulus histogram - SC synchronization coefficient - 0.5, 1, 2, 3, 10 Hz stimulus trains     

Monoenyl hydrocarbons in female body wax of the yellow peach moth as synergists of aldehyde pheromone components

The non-polar components of female body wax and pheromone gland extracts of the yellow peach moth synergistically enhanced male behavioral responses from close to pheromone sources in wind tunnel tests when mixed with an aldehyde pheromone blend. When the non-polar fractions (NPFs) of female body wax were further separated by column chromatography, synergistic activities were found in the 3 and 50% ether in hexane fractions, and they additively increased male responses. The main components of the first fraction were (Z)-9-tricosene, (Z)-9-pentacosene, (Z)-9-heptacosene, (Z)-9-nonacosene and (Z)-9-hentriacontene. Only (Z)-9-heptacosene showed a significant synergistic effect in enhancing male responses, but the other components had no effect. A mixture of the five monoenyl hydrocarbons lost activity at lower doses than 5 ng. Natural ratios of these hydrocarbons in the female body wax and pheromone gland extracts were similar, but the amount of (Z)-9-heptacosene in the female body wax was significantly higher than in the pheromone gland extracts. We conclude that (Z)-9-heptacosene increases male responses to aldehyde pheromones, and unknown component(s) in the 50% ether in the hexane fraction are required for full synergistic enhancement by the NPFs of the female body wax and the pheromone gland extracts.     

Chirp rate variability in male song ofEphippigerida taeniata (Orthoptera: Ensifera)

Variability in the chirp rate of the male song of the ephippigerine speciesEphippigerida taeniata during intraspecific communication was investigated in the laboratory. Conspecific chirps were used as auditory stimuli. The stimulus rate was controlled by computer. Experiments were carried out at 19, 27, and 35°C. Acoustically isolated males ofE. taeniata sang with a relatively constant chirp rate, which depended on the ambient temperature. Chirp rate significantly increased with rising temperature from 19 to 27°C, whereas at 35°C the chirp rate did not differ significantly from that at 27°C. Male chirp rates were affected by stimulus rates. Males significantly increased their chirp rate in response to increasing stimulus rates at temperatures of 19 and 27°C. At 35°C the increase in the chirp rate was not significant. At 27 and 35°C males sang with a higher chirp rate than the stimulus rate within a certain range. Evaluating stimulus and response chirp pattern when the males increased their chirp rate in response to the stimulus rate showed that an alternation pattern was established. More than 50% of the male chirps occurred at a characteristic time range at around 40% of the interstimulus interval, which was slightly affected by temperature.     

Effects of temperature on a moth auditory receptor

Measurements of the thoracic temperature and recordings of the spike activity of the most sensitive auditory receptor (A1 cell) were made in Empyreuma pugione (Arctiidae, Ctenuchinae). The temperature range tested (19–36 °C) is relevant for the behavior and ecology of this species. Experiments were performed during the hours of maximal flying activity in the wild: sunrise and sunset. The thoracic temperature during rest reflects that of the surrounding air; there is an increase of 3–4 °C immediately after ceasing free flying in the laboratory. The spike activity of the tympanic organ was recorded with a stainless-steelhook electrode placed beneath the tympanic nerve in the mesothorax. The A1 cell activity was studied without acoustic stimulation (spontaneous) and in response to 35-kHz acoustic pulses of 20, 40, or 100 ms duration. At all of these durations A1 cell response to saturating stimulus was analysed, while with 40-ms pulses different stimulus intensities were used (20–90 dB SPL in 10-dB steps). The number of action potentials per pulse, mean spike rate, maximal instantaneous discharge, and latency period depend strongly on air temperature, while the variation coefficients of the interspike intervals during the responses were not temperature dependent and vary non-monotonically with stimulus intensity. During responses to a saturating stimulus, the stimulus duration does not affect the activation energy, calculated from an Arrhenius plot, of different physiological features. Adaptation, studied in the responses to 100-ms pulses, is also temperature dependent. This phenomenon has two components, each of which shows different activation energies, suggesting a different membrane origin. High stimulus intensity (90 dB SPL) significantly affects the activation energy of the action potentials and mean spike rate, while the activation energy, of the maximal instantaneous discharge and latency period do not show this strong dependency. The spontaneous A1 cell spike rate varies with temperature, as does the value of the mode of the relative frequency distribution of the interspike interval. The activation energy of the spike rates measured at A1 cell responses to saturating stimuli is in good agreement with that described in amphibian innerear hair cells. It is suggested that this moth auditory receptor cell also has mechanosensitive protein channels.Abbreviations AP/p action potentials per pulse - AP/s action potentials per second - CI confidence interval - E _a activation energy - ISI interspike interval - SD standard deviation - VC variation coefficient     

Decyl-thio-trifluoropropanone, a competitive inhibitor of moth pheromone receptors

An earlier study (Pophof 1998) showed that the esterase inhibitor decyl-thio-trifluoropropanone inhibited the responses of two receptor neurons of the moth Antheraea polyphemus tuned to straight-chain pheromone components, an acetate and an aldehyde, respectively. Here we report that decyl-thio-trifluoropropanone also inhibited the responses of two pheromone receptor neurons of Bombyx mori to bombykol and bombykal. In contrast, decyl-thio-trifluoropropanone activated receptor neurons of the moth Imbrasia cytherea tuned to the pheromone component (Z)-5-decenyl 3-methyl-butanoate. However, decyl-thio-trifluoropropanone did not affect the responses of two receptor neurons of B. mori females specialized to the plant volatiles benzoic acid and linalool, respectively. These results indicate that decyl-thio-trifluoropropanone, besides inhibiting the sensillar esterase, interferes with proteins involved specifically in the excitation of pheromone receptor neurons. In binding studies with radiolabelled decyl-thio-trifluoropropanone, the inhibitor was bound by the pheromone-binding protein of A. polyphemus. However, the amount of decyl-thio-trifluoropropanone causing response inhibition was 300 times lower than the amount of pheromone-binding protein present in the sensilla. Since the amount of decyl-thio-trifluoropropanone adsorbed corresponded to about the maximum number of receptor molecules calculated per sensillum, we expect that decyl-thio-trifluoropropanone, probably in complex with pheromone-binding protein, competitively inhibits the pheromone receptor molecules. Accepted: 8 January 2000     

Sex pheromone of the red clover casebearer moth,Coleophora deauratella,an invasive pest of clover in Canada

The red clover casebearer, Coleophora deauratella Lienig & Zeller (Lepidoptera: Coleophoridae), is an invasive pest of Trifolium species (Fabaceae) in Canada. We identified candidate sex pheromone components from female pheromone gland extracts using coupled gas chromatographic–electroantennographic analysis detection. Three compounds elicited an electrophysiological response from antennae and were identified as: (Z)‐7‐dodecenyl acetate, (Z)‐5‐dodecenyl acetate, and (Z)‐7‐dodecen‐1‐ol. Field tests of the candidate pheromone components revealed that males were attracted to a binary mixture of (Z)‐7‐dodecenyl acetate and (Z)‐5‐dodecenyl acetate. Male moth trap capture was greatest in traps baited with lures containing 100:10 or 100:20 ratios of these pheromone components, respectively. Trap capture was reduced when (Z)‐5‐dodecenyl acetate was present below 10 or above 20% of (Z)‐7‐dodecenyl acetate. Equal numbers of male moths were captured in traps baited with 10, 100, and 1 000 μg of the attractive binary mixture. These findings allow for the development of a pheromone‐based monitoring system for this invasive pest of clover in Canada.     

Variations of opisthaptoral hard parts of Gyrodactylus salaris Malmberg, 1957 (Monogenea: Gyrodactylidae) on parr of Atlantic Salmon Salmo salar L. in laboratory experiments

On parr of Atlantic salmon Salmo salar L. in laboratory experiments at different water temperatures, opisthaptoral hard parts of Gyrodactylus salaris Malmberg, 1957 differed in size. Their shape, on the other hand, varied only slightly. The opisthaptoral hard parts were largest at the lowest water temperatures and decreased in size with increasing water temperatures (1.5°C to 20.0°C). At both 18.0°C and 20.0°C the opisthaptoral hard parts of G. salaris were smaller than has been found under natural conditions. In the experiments, the mean size of the opisthaptor in the G. salaris micropopulation at each water temperature gradually increased or decreased compared to the mean size at the start of the experiments. At the end of the experiments the mean size of the opisthaptoral hard parts in the G. salaris micropopulations showed a significant regression to the different water temperatures.     

Effect of rearing temperature and larval density on larval survival, age at pupation and adult size of Anopheles gambiae

The effects of temperature and larval density on survival of larvae, growth rate, age at pupation, and adult size (measured as wing length and dry weight) of laboratory-reared Anopheles gambiae (Diptera: Culicidae) were studied. Larvae were reared at three temperatures (24, 27 and 30°C) and three densities (0.5, 1 and 2 larvae/cm²). The effects of density and temperature strongly interacted to determine the mosquitoes' life-history parameters. Survival was highest at the intermediate temperature of 27°C. The differences between the temperatures increased with increasing density. At 30°C survival decreased as density increased, but at 27°C increasing density led to higher survival. Age at pupation increased as temperature decreased from 30°C to 24°C and as density decreased from 2 to 0.5 larvae/cm². Adult size also increased as temperature decreased, but showed a negative correlation with density only at 27°C. In contrast, at 24°C and 30°C a decrease in density led to a decrease in adult size. Growth rate showed a similar pattern. At 27°C growth rate decreased as density increased, but at other temperatures the opposite trend was observed.     

Pheromone receptor cells in the male moth Manduca sexta

Three types of pheromone receptor cells have been identified by electrophysiological recording from single antennal sensilla trichodea of the male sphinx moth Manduca sexta. These cells responded best to the pheromone components (E,Z)-10,12-hexadecadienal (type A receptor cell), (E,E,Z)-10,12,14-hexadecatrienal (type B), and (E,E,E)-10,12,14-hexadecatrienal (type C). Cell type B also responded to (E,Z)-11,13-pentadecadienal, which has been used experimentally as a pheromone substitute. In recordings from 20 trichoid hairs, 17 were found to be innervated by one cell of type A and one of type B; 3 trichoid hairs had cell types A and C.     

Antennal response of codling moth males, Cydia pomonella L. (Lepidoptera: Tortricidae), to the geometric isomers of codlemone and codlemone acetate

Single sensillum recordings from Cydia pomonella male antennae showed three different types of receptor neurons. The most abundant type was most sensitive to the main pheromone compound (E,E)-8,10-dodecadienol, while its response to the geometric isomers E,Z, Z,E and Z,Z was comparable to a tenfold lower dose of (E,E)-8,10-dodecadienol. This neuron type also responded to the four behaviorally antagonistic isomers of (Δ,Δ)-8,10-dodecadienyl acetate, among which it was most sensitive to the E,E isomer. Cross-adaptation studies showed that these compounds were all detected by the same receptor neuron type. Receptor neurons specifically tuned to (E,Z) or (Z,Z)-8,10-dodecadienol were not found, although these two compounds are behaviorally active. A second type of receptor neuron responded to all isomers of (Δ,Δ)-8,10-dodecadienyl acetate and was most sensitive to the E,E isomer. This neuron type did not respond to any of the isomers of (Δ,Δ)-8,10-dodecadienol. A third receptor neuron type was highly sensitive to the plant compound α-farnesene. The finding that the receptor neuron type tuned to the main pheromone compound responded even to strong behavioral antagonists aids the interpretation of ongoing behavioral studies for the development of the mating disruption technique in codling moth. Accepted: 3 March 2000     

Neurons discovered in male Helicoverpa zea antennae that correlate with pheromone-mediated attraction and interspecific antagonism

Responses of single receptor neurons in the antennae of male Helicoverpa zea to sex pheromone components and to behavioral antagonists were recorded using a cut-sensillum extracellular recording technique. Three types of sensilla were identified from sampling 325 male-specific sensilla trichodea located at the lateral edge of antennomeres. The majority of these sensilla (71%) contained a receptor neuron tuned to the principal sex pheromone component (Z)-11-hexadecenal. A second sensillar type (10%) contained a receptor neuron that responded only to (Z)-9-tetradecenal. A third sensillar type (19%) contained a large-spiking neuron tuned to the secondary pheromone component (Z)-9-hexadecenal, but this neuron also could be stimulated to equivalent spike frequencies by the same emitted amounts of (Z)-9-tetradecenal. A smaller-spiking neuron in this sensillar type responded to two compounds known to act only as behavioral antagonists, (Z)-11-hexadecen-1-ol and (Z)-11-hexadecenyl acetate, and to (Z)-9-tetradecenal. Cross-adaptation studies confirmed the presence of one large- and one small-spiking neuron in the third sensillar type. Dose-response studies correlated to collected stimuli amounts showed that the large-spiking neuron in the third sensillar type was equally tuned to (Z)-9-hexadecenal and (Z)-9-tetradecenal, whereas the smaller-spiking neuron was far more sensitive to (Z)-11-hexadecen-1-ol and to (Z)-11-hexadecenyl acetate than to (Z)-9-tetradecenal. Accepted: 29 September 1997