Neil J. Vickers

Mechanisms of animal navigation in odor plumes

Chemical signals mediate many of lifes processes. For organisms that use these signals to orient and navigate in their environment, where and when these cues are encountered is crucial in determining behavioral responses. In air and water, fluid mechanics impinge directly upon the distribution of odorous molecules in time and space. Animals frequently employ behavioral mechanisms that allow them to take advantage of both chemical and fluid dynamic information in order to move toward the source. In turbulent plumes, where odor is patchily distributed, animals are exposed to a highly intermittent signal. The most detailed studies that have attempted to measure fluid dynamic conditions, odor plume structure, and resultant orientation behavior have involved moths, crabs, and lobsters. The behavioral mechanisms employed by these organisms are different but generally integrate some form of chemically modulated orientation (chemotaxis) with a visual or mechanical assessment of flow conditions in order to steer up-current or upwind (rheo- or anemo-taxis, respectively). Across-stream turns are another conspicuous feature of odor-modulated tracks of a variety of organisms in different fluid conditions. In some cases, turning is initiated by detection of the lateral edges of a well-defined plume (crabs), whereas in other animals turning appears to be steered according to an internally generated program modulated by odor contacts (moth counterturning). Other organisms such as birds and fish may use similar mechanisms, but the experimental data for these organisms is not yet as convincing. The behavioral strategies employed by a variety of animals result in orientation responses that are appropriate for the dispersed, intermittent plumes dictated by the fluid-mechanical conditions in the environments that these different macroscopic organisms inhabit.

Odour-plume dynamics influence the brain's olfactory code

The neural computations used to represent olfactory information in the brain have long been investigated. Recent studies in the insect antennal lobe suggest that precise temporal and/or spatial patterns of activity underlie the recognition and discrimination of different odours, and that these patterns may be strengthened by associative learning. It remains unknown, however, whether these activity patterns persist when odour intensity varies rapidly and unpredictably, as often occurs in nature. Here we show that with naturally intermittent odour stimulation, spike patterns recorded from moth antennal-lobe output neurons varied predictably with the fine-scale temporal dynamics and intensity of the odour. These data support the hypothesis that olfactory circuits compensate for contextual variations in the stimulus pattern with high temporal precision. The timing of output neuron activity is constantly modulated to reflect ongoing changes in stimulus intensity and dynamics that occur on a millisecond timescale.

A comparison of responses from olfactory receptor neurons of Heliothis subflexa and Heliothis virescens to components of their sex pheromone

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons in sensilla trichodea on male antennae of the heliothine species Heliothis subflexa and the closely related congener H. virescens. A large percentage of sensilla (72% and 81%, respectively, of all sensilla sampled) contained a single odor-responsive receptor neuron tuned to the major pheromone component of both species, Z-11-hexadecenal. A second population of sensilla on H. subflexa antennae (18%) housed receptor neurons that were tuned to Z-9-hexadecenal but also responded with less sensitivity to Z-9-tetradecenal. A similar population of sensilla (4%) on H. virescens male antennae housed receptor neurons that were shown to be tuned specifically only to Z-9-tetradecenal, with no response to even high dosages of Z-9-hexadecenal. A third population of sensilla (comprising 8% and 16% of the sensilla sampled in H. subflexa and H. virescens, respectively) housed two olfactory receptor neurons, one of which was tuned to Z-11-hexadecenyl acetate and the other tuned to Z-11-hexadecenol. In H. subflexa the Z-11-hexadecenyl acetate-tuned neuron also responded to Z-9-tetradecenal with nearly equivalent sensitivity. The behavioral requirements of males of these two species for distinct pheromonal blends was, therefore, reflected by the subtle differences in the tuning properties of antennal olfactory receptor neurons.

Central Processing of Natural Odor Mixtures in Insects

In nature, virtually all olfactory stimuli are mixtures of many single odorants. Behavioral experiments repeatedly have demonstrated that an animal’s olfactory system is capable of discriminating behaviorally relevant from irrelevant odor mixtures. However, the sensory mechanisms that underlie such discriminative capability remain elusive. The limited anatomical and physiological evidence collected from both insect and vertebrate models that pertains to this topic is scattered in the literature dating back to early 1980s. Thus, a synthesis of this information that includes more recent findings is needed in order to provide a basis for probing the fundamental question from a new angle. In this review, we discuss several proposed models for mixture processing, along with experimental data gathered from both the initial stage of olfactory processing (i.e., antennal lobe in insects or olfactory bulb in vertebrates) and higher areas of the brain, with an emphasis on how the lateral circuits in the antennal lobe or olfactory bulb may contribute to mixture processing. Based on empirical data as well as theoretical modeling, we conclude that odor mixtures may be represented both at the single-neuron level and at the population level. The difference between these two types of processing may reside in the degree of plasticity, with the former being hard-wired and the latter being more subjected to network modulation.

Sexual isolation of male moths explained by a single pheromone response QTL containing four receptor genes

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.

Physiology and glomerular projections of olfactory receptor neurons on the antenna of female Heliothis virescens (Lepidoptera: Noctuidae) responsive to behaviorally relevant odors

The neurophysiology and antennal lobe projections of olfactory receptor neurons housed within short trichoid sensilla of female Heliothis virescens F. (Noctuidae: Lepidoptera) were investigated using a combination of cut-sensillum recording and cobalt-lysine staining techniques. Behaviorally relevant odorants, including intra- and inter-sexual pheromonal compounds, plant and floral volatiles were selected for testing sensillar responses. A total of 184 sensilla were categorized into 25 possible sensillar types based on odor responses and sensitivity. Sensilla exhibited both narrow (responding to few odors) and broad (responding to many odors) response spectra. Sixty-six percent of the sensilla identified were stimulated by conspecific odors; in particular, major components of the male H. virescens hairpencil pheromone (hexadecanyl acetate and octadecanyl acetate) and a minor component of the female sex pheromone, (Z)-9-tetradecenal. Following characterization of the responses, olfactory receptor neurons within individual sensilla were stained with cobalt lysine (N=39) and traced to individual glomeruli in the antennal lobe. Olfactory receptor neurons with specific responses to (Z)-9-tetradecenal, a female H. virescens sex pheromone component, projected to the female-specific central large female glomerulus (cLFG) and other glomeruli. Terminal arborizations from sensillar types containing olfactory receptor neurons sensitive to male hairpencil components and plant volatiles were also localized to distinct glomerular locations. This information provides insight into the representation of behaviorally relevant odorants in the female moth olfactory system.

Trapping Phyllophaga spp. (Coleoptera: Scarabaeidae: Melolonthinae) in the United States and Canada using sex attractants.

Abstract The sex pheromone of the scarab beetle, Phyllophaga anxia, is a blend of the methyl esters of two amino acids, L-valine and L-isoleucine. A field trapping study was conducted, deploying different blends of the two compounds at 59 locations in the United States and Canada. More than 57,000 males of 61 Phyllophaga species (Coleoptera: Scarabaeidae: Melolonthinae) were captured and identified. Three major findings included: (1) widespread use of the two compounds [of the 147Phyllophaga (sensu stricto) species found in the United States and Canada, males of nearly 40% were captured]; (2) in most species intraspecific male response to the pheromone blends was stable between years and over geography; and (3) an unusual pheromone polymorphism was described from P. anxia. Populations at some locations were captured with L-valine methyl ester alone, whereas populations at other locations were captured with L-isoleucine methyl ester alone. At additional locations, the L-valine methyl ester-responding populations and the L-isoleucine methyl ester-responding populations were both present, producing a bimodal capture curve. In southeastern Massachusetts and in Rhode Island, in the United States, P. anxia males were captured with blends of L-valine methyl ester and L-isoleucine methyl ester. Resumen La feromona sexual del escarabajo, Phyllophaga anxia, es una mezcla de los ésteres metílicos de dos aminoácidos, L-valina y L-isoleucina. Se condujo un estudio de campo usando diferentes mezclas de los dos componentes en 59 sitios de Estados Unidos y Canada. Más de 57,000 machos de 61 especies dePhyllophaga fueron capturados e identificados. Tres de los resultados más importantes incluyen: (1) el extenso uso de los dos componentes [de las 147 especies de Phyllophaga (sensu stricto), en Estados Unidos y Canada, fueron capturados machos de cerca del 40% de ellas.]; (2) para la mayoría de las especies, la respuesta intraespecífica de los machos a las combinaciones de los dos aminoácidos fue consistente entre años diferentes, y en todos los sitios geográficos; y (3) un inusual polymorfismo de la feromona fue descrito para P. anxia. Poblaciones de algunos sitios fueron atrapados sólo con valina, mientras que poblaciones de otros sitios fueron atrapados sólo con isoleucina. También se encontraron sitios donde las poblaciones responden a ambos componentes, valina e isoleucina, produciendo una curva de captura bimodal. En el sureste del estado de Massachusetts y en Rhode Island, en Estados Unidos, machos de P. anxia fueron atrapados en trampas con mezclas de valina e isoleucina.

Defining a Synthetic Pheromone Blend Attractive to Male Heliothis subflexa Under Wind Tunnel Conditions

Heliothis subflexa males were flown in a wind tunnel to a variety of combinations of synthetic pheromone components admixed on a filter paper disk. Blends containing (Z)-11-hexadecenal (Z11–16:Ald, 1000 ng), (Z)-9-hexadecenal (Z9–16:Ald, 500 ng) and (Z)-11-hexadecenol (Z11–16:OH, 10–500 ng) elicited upwind flight and source contact in 52–69% of males. All these compounds have previously been isolated and identified from female H. subflexa gland extracts and volatile pheromone emissions. Males were not attracted by blends in which Z9–16:Ald was omitted (0% source contact). Similarly, blends lacking Z11–16:OH were unattractive to male H. subflexa (3% or less source contact). Males were extremely sensitive to the presence of Z11–16:OH; however, responding in high numbers (57–69% source contact) to blends containing a dosage of 1% (10 ng) or greater Z11–16:OH. Males were unresponsive to blends in which Z9–16:Ald was replaced with a variety of dosages of (Z)-9-tetradecenal, a secondary component of a closely-related congeneric species, Heliothis virescens. Another compound present in the blend emitted by conspecific females, (Z)-11-hexadecenyl acetate (Z11–16:Ac), did not inhibit H. subflexa males (69% source contact) when added to the three-component mixture (1:0.5:0.1) at a ratio of 0.1 (100 ng) with respect to Z11–16:Ald. These results indicate that Z9–16:Ald and Z11–16:OH are required in addition to Z11–16:Ald to elicit significant levels of upwind flight in H. subflexa males. The effects of Z11–16:Ac are more subtle, but at the dosage tested in these experiments, this compound does not have an antagonistic effect on upwind flight and source location by H. subflexa males.

Pheromone-Mediated Flight in Moths

Several explanations for the existence of zigzagging upwind flight in male moths responding to sex pheromone have been given over the course of research on this topic (e.g. Kennedy 1983; Preiss and Kramer 1986b; Baker and Haynes 1987; David and Kennedy 1987; David and Birch 1989; Baker 1990; Witzgall and Arn 1990; Willis and Arbas 1991a). Although a debate about the superficial shape of the flight tracks appears to be over-reductionistic, the root of the discussion involves the mechanisms that the moths use to maneuver and reach the source of pheromone in wind. As such, it is a very important debate. The behavioral mechanisms need to be precisely understood if we are to make sense of the underlying neuronal responses at the sensory, central nervous system, and motor levels and create robust neuroethological knowledge about this powerful and agriculturally important biological process called attraction.

Inheritance of Olfactory Preferences II. Olfactory Receptor Neuron Responses from Heliothis subflexa × Heliothis virescens Hybrid Male Moths

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons (ORNs) in sensilla trichodea on male antennae of hybrids formed mainly by crossing female Heliothis subflexa with male Heliothis virescens (‘SV hybrids’). We recorded from the A-, B-, and C-type sensilla trichodea, with the latter two types housing ORNs exhibiting response profiles to different pheromone components that we had previously found to be characteristic for each species. For both the B- and the C-type SV hybrid sensilla, most of the ORNs exhibited a spike amplitude and ORN co-compartmentalization within sensilla that more strongly resembled the ORNs of parental H. subflexa rather than those of H. virescens. The overall mean dose-response profiles of the ORNs in hybrid C- and B-type sensilla were intermediate between those of the H. virescens and H. subflexa parental type ORNs. However, not all hybrid ORNs were intermediate in their tuning spectra, but rather ranged from those that closely resembled H. subflexa or H. virescens parental types to those that were intermediate, even on the same antenna. The most noteworthy shift in ORN responsiveness in hybrid males was an overall increase in sensitivity to Z9–14:Ald exhibited by Z9–16:Ald-responsive ORNs. Heightened cross-responsiveness to Z9–14:Ald by hybrid ORNs correlates well with observed behavioral cross-responsiveness of hybrids in which Z9–14:Ald could substitute for Z9–16:Ald in the pheromone blend, a behavior not observed in parental types. The hybrid ORN shifts involving greater sensitivity to Z9– 14:Ald also correlate well with studies of hybrid male antennal lobe interneurons that exhibited a shift toward greater cross-responsiveness to Z9–14:Ald and Z9– 16:Ald. We propose that the differences between parental H. virescens, H. subflexa, and SV hybrid male pheromone ORN responsiveness to Z9–16:Ald and Z9–14:Ald are most logically explained by an increased or decreased co-expression of two different odorant receptors for each of these compounds on the same ORN.