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Olfactory projection neurons convey information from the insect antennal lobe (AL) to higher brain centers. Previous reports have demonstrated that pheromone-responsive projection neurons with cell bodies in the moth medial cell cluster (mcPNs) predominantly have dendritic arborizations in the sexually dimorphic macroglomerular complex (MGC) and send an axon from the AL to the calyces of the mushroom body (CA) as well as the lateral horn (LH) of the protocerebrum via the medial AL tract. These neurons typically exhibit a narrow odor tuning range related to the restriction of their dendritic arbors within a single glomerulus (uniglomerular). In this study, we report on the diverse physiological and morphological properties of a group of pheromone-responsive olfactory projection neurons with cell bodies in the AL lateral cell cluster (MGC lcPNs) of two closely related moth species. All pheromone-responsive lcPNs appeared to exhibit "basket-like" dendritic arborizations in two MGC compartments and made connections with various protocerebral targets including ventrolateral and superior neuropils via projections primarily through the lateral AL tract and to a lesser extent the mediolateral antennal lobe tract. Physiological characterization of MGC lcPNs also revealed a diversity of response profiles including those either enhanced by or reliant upon presentation of a pheromone blend. These responses manifested themselves as higher maximum firing rates and/or improved temporal resolution of pulsatile stimuli. MGC lcPNs therefore participate in conveying diverse olfactory information relating to qualitative and temporal facets of the pheromone stimulus to a more expansive number of protocerebral targets than their mcPN counterparts., (© 2019 Wiley Periodicals, Inc.)

Behavioral responses to odors rely first upon their accurate detection by peripheral sensory organs followed by subsequent processing within the brain's olfactory system and higher centers. These processes allow the animal to form a unified impression of the odor environment and recognize combinations of odorants as single entities. To investigate how interactions between peripheral and central olfactory pathways shape odor perception, we transplanted antennal imaginal discs between larval males of two species of moth Heliothis virescens and Heliothis subflexa that utilize distinct pheromone blends. During metamorphic development olfactory receptor neurons originating from transplanted discs formed connections with host brain neurons within olfactory glomeruli of the adult antennal lobe. The normal antennal receptor repertoire exhibited by males of each species reflects the differences in the pheromone blends that these species employ. Behavioral assays of adult transplant males revealed high response levels to two odor blends that were dissimilar from those that attract normal males of either species. Neurophysiological analyses of peripheral receptor neurons and central olfactory neurons revealed that these behavioral responses were a result of: 1. the specificity of H. virescens donor olfactory receptor neurons for odorants unique to the donor pheromone blend and, 2. central odor recognition by the H. subflexa host brain, which typically requires peripheral receptor input across 3 distinct odor channels in order to elicit behavioral responses.

Long distance sexual communication in moths has fascinated biologists because of the complex, precise female pheromone signals and the extreme sensitivity of males to specific pheromone molecules. Progress has been made in identifying some genes involved in female pheromone production and in male response. However, we have lacked information on the genetic changes involved in evolutionary diversification of these mate-finding mechanisms that is critical to understanding speciation in moths and other taxa. We used a combined quantitative trait locus (QTL) and candidate gene approach to determine the genetic architecture of sexual isolation in males of two congeneric moths, Heliothis subflexa and Heliothis virescens. We report behavioral and neurophysiological evidence that differential male responses to three female-produced chemicals (Z9-14:Ald, Z9-16:Ald, Z11-16:OAc) that maintain sexual isolation of these species are all controlled by a single QTL containing at least four odorant receptor genes. It is not surprising that pheromone receptor differences could control H. subflexa and H. virescens responses to Z9-16:Ald and Z9-14:Ald, respectively. However, central rather than peripheral level control over the positive and negative responses of H. subflexa and H. virescens to Z11-16:OAc had been expected. Tight linkage of these receptor genes indicates that mutations altering male response to complex blends could be maintained in linkage disequilibrium and could affect the speciation process. Other candidate genes such as those coding for pheromone binding proteins did not map to this QTL, but there was some genetic evidence of a QTL for response to Z11-16:OH associated with a sensory neuron membrane protein gene.

The western boxelder bug (BEB), Boisea rubrolineata (Heteroptera: Rhopalidae), is a specialist herbivore of boxelder trees, Acer negundo. We tested the hypothesis that BEBs use semiochemicals to locate host trees. Headspace volatiles from trees bearing staminate inflorescences ("staminate trees") and from trees bearing pistillate inflorescences ("pistillate trees") were collected throughout the season and bioassayed in Y-tube olfactometer experiments. Headspace extracts of early-season, pollen-bearing staminate trees and midseason pistillate trees with mature samaras (seed pods) attracted female and male BEBs. By using coupled gas chromatographic-electroantennographic detection and gas chromatography-mass spectrometry, we identified and tested a five-component synthetic blend of candidate semiochemicals (hexanol, pentyl acetate, phenylacetonitrile, 2-phenethyl acetate, and trans-nerolidol). This blend attracted females, males, and fifth-instar nymphs. Phenylacetonitrile by itself was as attractive as the five-component blend to both adults and nymphs. By responding to phenylacetonitrile emitted by pollen-bearing staminate trees and pistillate trees with maturing seeds, BEBs appear to track and exploit the availability of nutrient-rich food sources, suggesting that the bugs' reproductive ecology is synchronized to the phenology of their host boxelder tree.

Terrestrial odor plumes have a physical structure that results from turbulence in the fluid environment. The rapidity of insect flight maneuvers within a plume indicates that their responses are dictated by fleeting (<1 s) rather than longer (>1 s) exposures to odor imposed by physical variables that distribute odor molecules in time and space. Even though encounters with pheromone filaments are brief, male moths responding to female-produced pheromones are remarkably able to extract information relating to the biological properties of these olfactory signals. These properties include the types of molecule present and their relative abundances. Thus, peripheral and central olfactory neurons are capable of representing these biological properties of a pheromone plume within the context of a temporally irregular and unpredictable signal. The mechanisms underlying olfactory processing of these signals with respect to their biological and physical properties are discussed in the context of a behavioral framework.

At low ambient temperature Helicoverpa zea male moths engage in warm-up behavior prior to taking flight in response to an attractive female pheromone blend. Male H. zea warm up at a faster rate when sensing the attractive pheromone blend compared with unattractive blends or blank controls (Crespo et al. 2012), but the mechanisms involved in this olfactory modulation of the heating rate during preflight warm-up are unknown. Here, we test three possible mechanisms for increasing heat production: 1) increased rate of muscle contraction; 2) reduction in mechanical movement by increased overlap in activation of the antagonistic flight muscles; and 3) increased activation of motor units. To test which mechanisms play a role, we simultaneously recorded electrical activation patterns of the main flight muscles (dorsolongitudinal and dorsoventral muscles), wing movement, and thoracic temperature in moths exposed to both the attractive pheromone blend and a blank control. Results indicate that the main mechanism responsible for the observed increase in thoracic heating rate with pheromone stimulation is the differential activation of motor units during each muscle contraction cycle in both antagonistic flight muscles. This additional activation lengthens the contracted state within each cycle and thus accounts for the greater heat production. Interestingly, the rate of activation (frequency of contraction cycles) of motor units, which is temperature dependent, did not vary between treatments. This result suggests that the activation rate is determined by a temperaturedependent oscillator, which is not affected by the olfactory stimulus, but activation of motor units is modulated during each cycle. [ABSTRACT FROM AUTHOR]

Reproductive isolation between sympatric, closely related species can be accomplished through a variety of pre-zygotic isolating mechanisms, including courtship-signaling behavior that involves pheromones. In the moths Heliothis virescens and H. subflexa, males display abdominal hairpencils (HP), which release volatile chemicals during courtship. In this study, we demonstrated that HP volatiles released by male H. subflexa function to improve mating success with conspecific females. Interspecific mating experiments were conducted to determine any influence of HP volatiles on species isolation. Female H. virescens and H. subflexa were observed during courtship with males of the other species, following either sham-operation or ablation of HPs, both with and without concurrent presentation of HP volatiles. Mating success was improved by co-presentation of HP extract from males of the same species during courtship. Ablation of HPs improved mating between H. subflexa females and H. virescens males. During interspecific matings, male H. virescens attempted copulation less frequently in the presence of H. virescens HP extract, though H. subflexa males were not affected by the presence of H. subflexa HP extract. This suggests that HP volatiles produced by males of each species may inhibit mating between species through effects on males ( H. virescens) and females ( H. subflexa). [ABSTRACT FROM AUTHOR]

Long distance sexual communication in moths has fascinated biologists because of the complex, precise female pheromone signals and the extreme sensitivity of males to specific pheromone molecules. Progress has been made in identifying some genesinvolved in female pheromone production and in male response. However, we have lacked information on the genetic changes involved in evolutionary diversification of these mate-finding mechanisms that is critical to understanding speciation in moths and other taxa. We used a combined quantitative trait locus (QTL) and candidate gene approach to determine the genetic architecture of sexual isolation in males of two congeneric moths, Heliothis sub flexa and Heliothis virescens. We report behavioral and neurophysiological evidence that differential male responses to three female-produced chemicals (Z9-14:Ald, Z9-16:Ald, Z11-16:OAc) that maintain sexual isolation of these species are all controlled by a single QTL containing at least four odorant receptor genes. It is not surprising that pheromone receptor differences could control H. sub flexa and H. virescens responses to 29-1 6:Ald and 29-14:Ald, r,espectively. However, central rather than peripheral level control over the positive and negative responses of H. sub flexa and H. virescens to Z11-16:OAc had been expected. Tight linkage of these receptor genes indicates that mutations altering male response to complex blends could be maintained in linkage disequilibrium and could affect the speciation process. Other candidate genes such as those coding for pheromone binding proteins did not map to this QTL but there was some genetic evidence of a QTL for response to Z11-16:OH associated with a sensory neuron membrane protein gene. [ABSTRACT FROM AUTHOR]

We used single-sensillum recordings to characterize male Heliothis subflexa antennal olfactory receptor neuron physiology in response to compounds related to their sex pheromone. The recordings were then followed by cobalt staining in order to trace the neurons' axons to their glomerular destinations in the antennal lobe. Receptor neurons responding to the major pheromone component, (Z)-11-hexadecenal, in the first type of sensillum, type-A, projected axons to the cumulus of the macroglomerular complex (MGC). In approximately 40% of the type-A sensilla, a colocalized receptor neuron was stained that projected consistently to the posterior complex 1 (PCx1), a specific glomerulus in an 8-glomerulus complex that we call the Posterior Complex (PCx). We found that receptor neurons residing in type-B sensilla and responding to a secondary pheromone component, (Z)-9-hexadecenal, send their axons to the dorsal medial glomerulus of the MGC. As in the type-A sensilla, we found a cocompartmentalized neuron within type-B sensilla that sends its axon to a different glomerulus of the PCx4. One neuron in type-C sensilla tuned to a third pheromone component, (Z)-11-hexadecenol, and a colocalized neuron responding to (Z)-11-hexadecenyl acetate projected their axons to the anteromedial and ventromedial glomeruli of the MGC, respectively. [ABSTRACT FROM AUTHOR]

Shifts in male preference for qualitatively different pheromone blends appear to have played a fundamental role in the divergence of olfactory communication and evolution of moth species. As an initial step in documenting the genetic complexity underlying such shifts, we characterized the behavioral responses of hybrid male moths created by mating two heliothine moth species, Heliothis subflexa and Heliothis virescens. Between 67 and 96% of hybrid males flew upwind and contacted the pheromone source when presented with a blend consisting of (Z)-11-hexadecenal (Z11–16:Ald), (Z)-9-hexadecenal (Z9–16:Ald), and (Z)-11-hexadecenol (Z11–16:OH) in a 1:0.5:0.1 ratio that has previously been shown to be attractive to H. subflexa males. In addition, an H. virescens blend of Z11–16:Ald and (Z)-9-tetradecenal (Z9–14:Ald) enhanced by the addition of Z11–16:OH (in a 1:0.05:0.1 mixture) was attractive to hybrid males (26–64% source contact), but significantly fewer males reached the odor source compared to the blend containing Z9–16:Ald. A blend in which the dosage of Z9–14:Ald was doubled, however, was equally attractive (75–77% source contact) as the Z9–16:Ald-containing blend. Consecutive presentation of two blends revealed that individual hybrid males responded equally well to blends containing either Z9–14:Ald or Z9–16:Ald. Together these results suggest that in addition to Z11–16:Ald, hybrid males: (1) required either Z9–16:Ald (likeH. subflexamales) or Z9–14:Ald (like H. virescens males); (2) required the presence of Z11–16:OH (H. subflexa dominant); (3) were not adversely affected by the presence of Z11–16:Ac (H. subflexa dominant). The behavioral response phenotype of hybrid males was therefore influenced by genetic factors inherited from both parental species. Copyright © 2006 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Pheromone-responsive olfactory interneurons were studied to determine the extent to which their physiological and morphological properties complemented the behavior and peripheral olfactory neurobiology observed in hybrid male moths created by interbreeding two species of heliothine moth, Heliothis virescens and Heliothis subflexa. Complete recordings were made from a total of 33 neurons, and 16 projection neurons (PNs) were subsequently stained with a fluorescent dye. Stained PNs tuned to pheromonal odorants had dendritic arborizations restricted to one of four olfactory glomeruli that together constituted the macroglomerular complex (MGC). As in parental males, PNs tuned to (Z)-11-hexadecenal always had an arbor in the cumulus, the largest of the MGC glomeruli. Previous neurophysiological investigations revealed that PNs with dendritic arbors restricted to the dorso-medial glomerulus (DM) of the MGC responded specifically to either (Z)-9-tetradecenal (Z9–14:Ald; H. virescens males) or (Z)-9-hexadecenal (Z9–16:Ald; H. subflexa males). Hybrid males, which responded equally well in wind tunnel tests to blends containing either Z9–14:Ald or Z9–16:Ald, had DM PNs that responded to both odorants. PNs specific for a third compound, (Z)-11-hexadecenol, required by hybrid males for behavioral activity were localized to the antero-medial MGC glomerulus (AM). Thus, neuronal activity across the cumulus, DM and AM glomeruli represented an attractive blend in hybrid males. Neurons tuned to (Z)-11-hexadecenyl acetate and Z9–14:Ald were restricted to a fourth, ventro-medial glomerulus. The across-glomerular pattern of activity associated with attractive pheromone blends was most similar to that of H. subflexa males, signifying a dominant effect of H. subflexa genes. These results indicate that the behavioral phenotype of hybrid males can be linked to underlying central olfactory characteristics. Copyright © 2006 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

In different moth species, the number and spatial arrangement of olfactory glomeruli in the antennal lobe (AL) vary widely, but the spatial map within a species is thought to be invariant, making it possible to identify single glomeruli across individuals. We investigated the relationship between the physiological tuning of pheromone-selective interneurons and their association with specific, identified glomeruli in the macroglomerular complex (MGC) of the noctuid moth, Heliothis subflexa. Three odorants that are required for pheromone-source location in this species were tested individually and in blends. Recordings from 27 pheromone-specific projection neurons (PNs) indicated that the majority (48%) were selectively activated by the major pheromone component of this species, Z-11-hexadecenal (Z11–16:Ald), with 33% primarily tuned to Z-9-hexadecenal and 19% to Z-11-hexadecenol. Intracellular staining revealed that the dendrites of PNs tuned to Z11–16:Ald always branched within the largest glomerulus of the MGC, the cumulus. Similarly, each of the other two classes of PN was associated with a different `satellite' glomerulus in the MGC. The spatial configuration of the four-glomerulus H. subflexa MGC was indistinguishable from that previously reported in the closely related species, Heliothis virescens. Hence, as these two species diverged, changes in the association of satellite MGC glomeruli with particular odorants have occurred without a measurable change in the anatomical arrangement of the glomerular array. [ABSTRACT FROM AUTHOR]

Summary Male moths compete to arrive first at a female releasing pheromone. A new study reveals that additional pheromone cues released only by younger females may prompt males to avoid them in favor of older but more fecund females. [ABSTRACT FROM AUTHOR]

The article reports on the results of mating disruption experiments during two seasons using (Z,E)-7,9,11-dodecatrienyl formate. This is a formate ester analog of the key aldehyde component of the sex pheromone of the carob moth, Ectomyelois ceratoniae. Results indicated that the pheromone disrupted mate location when it was initially deployed. However, the effect declined between deployments as the disruptant was depleted under harsh field conditions.