Inheritance of olfactory preferences II. Olfactory receptor neuron responses from Heliothis subflexa x 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.

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.

Unusual pheromone receptor neuron responses in heliothine moth antennae derived from inter-species imaginal disc transplantation.

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons housed in sensilla trichodea along the adult antennae arising from transplantation of the antennal imaginal discs between larval male Helicoverpa zea and Heliothis virescens. The olfactory receptor neurons from the majority of type C sensilla sampled on transplanted antennae displayed response characteristics consistent with those of the species that donated the antennae. However, some of the sensilla type C sampled in either transplant type contained olfactory receptor neurons that responded in a manner typical of the recipient species or other neurons that have not previously been found in the type C sensilla of either species. The single-cell data help to explain behavioral results showing that some transplant males do fly upwind to both species' pheromone blends, an outcome not expected based on known antennal sensory phenotypes. Our results suggest that host tissue can influence antennal olfactory receptor neuron development, and further that because of a common phylogenetic ancestry the donor tissue has the genetic capability to produce a variety of sensillar and receptor types.

Host plant volatiles synergize responses of sex pheromone-specific olfactory receptor neurons in male Helicoverpa zea.

Single-cell electrophysiological recordings were obtained from olfactory receptor neurons in antennal trichoid sensilla of male corn earworm, Helicoverpa zea. Spontaneous activity of the neuron specific for the major component ( Z)-11-hexadecenal, the conspecific female-emitted sex pheromone, was not affected by exposure to host plant volatiles. However, stimulations with binary mixtures of a threshold dosage of the pheromone component and increasing dosages of either linalool or ( Z)-3-hexenol significantly synergized the pheromone-specific neuron's firing rates compared with responses to the major pheromone component alone. Cross-adaptation studies confirmed that the enhanced impulses originated from the pheromone-component-tuned neuron. Because plant volatiles do not stimulate the pheromone-specific neuron when presented alone, the pheromone plus host odor blend would be interpreted as containing more pheromone than it actually does when processed by the pheromone-processing portion of the antennal lobe.

Functional morphology of antennal chemoreceptors of the parasitoid Microplitis croceipes (Hymenoptera: Braconidae).

The specialist parasitoid, Microplitis croceipes (Cresson) (Hymenoptera: Braconidae), uses chemical cues from plants damaged by herbivore-feeding and also plant by-products in host location and acceptance. These chemicals are detected by the wasp's antennae. We conducted scanning and transmission electron microscopy studies of male and female M. croceipes antennae and detected no distinct morphological differences in the chemoreceptors between the sexes. Male antennae are approximately twice as long as female antennae. We found five morphological sensillar types in both sexes: sensilla (s) trichodea were the most abundant and distributed over the whole antenna; s. chaetica were present in low numbers only on the scape and pedicel; and s. basiconica, s. coeloconica and elongated s. placodea were found only on the flagellum. Ultrastructural investigations revealed pore systems on s. basiconica and s. placodea. In s. placodea, sensory neurons run parallel to the longitudinal axis of the sensilla with terminal pores. We recorded responses from single olfactory receptor neurons in s. placodea to two plant-emitted volatiles, cis-3-hexenol and ocimene, and two anthropogenic compounds, cyclohexanone and 2-diisopropylaminoethanol. Male receptor neurons were more sensitive than those of females with significantly higher spike frequency being registered from neurons in males.

Fine structure and distribution of antennal sensilla of the desert locust, Schistocerca gregaria (Orthoptera: Acrididae).

The fine structure and distribution of various types of antennal sensilla in three nymphal stages and in adults of both solitary-reared (solitary) and crowd-reared (gregarious) phases of the desert locust, Schistocerca gregaria, were investigated by scanning and transmission electron microscopy. Four types of sensilla were identified: sensilla basiconica, s. trichodea, s. coeloconica and s. chaetica. S. basiconica contain up to 50 sensory neurons, each of which displays massive dendritic branching. The sensillar wall is penetrated by a large number of pores. In contrast, s. trichodea contain one to three sensory neurons that branch to give five or six dendrites in the sensillar lumen; the sensillum wall is penetrated by relatively few pores. The s. coeloconica are situated in spherical cuticular pits on the antennal surface. The s. coeloconica are of two types: one type contains one to three sensory neurons with double sensillar walls penetrated by slit-like pores, whereas the second type contains four sensory neurons with non-porous double sensillar walls. The s. chaetica have a flexible socket and a thick non-porous sensillum wall and contain four sensory neurons that send unbranched dendrites to a terminal pore. A fifth sensory neuron of the s. chaetica terminates in a tubular body at the base of the hair. S. basiconica and coeloconica are normally distributed over the entire antennal flagellum, with a concentration in the middle segments; s. trichodea have three areas of concentration on the 5th, 10th and 14th flagellar segments. Sensilla chaetica are most abundant on the terminal segment. Locusts raised in solitary conditions have more olfactory sensilla (s. basiconica and s. coeloconica) than crowd-reared locusts. The difference in sensillar numbers is more evident in adults than in nymphs. These results suggest that differences in the odour-mediated behaviour of nymphs and adults, and between the phases of S. gregaria, may be attributable to differences at the sensory input level.

Antennal lobe projection patterns of olfactory receptor neurons involved in sex pheromone detection in Spodoptera littoralis (Lepidoptera: Noctuidae).

Pheromone-specific receptor neurons in male and female cotton leafworms, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) were typed physiologically and traced into the antennal lobe using cobalt lysine as a marker. In male S. littoralis, the macroglomerular complex (MGC), which is responsible for the initial integration of information concerning sex pheromone components, contains three glomerular compartments as revealed in a morphological study. No such specialized structure was seen in the female. In the male, olfactory receptor neurons that responded selectively to stimulation with the major sex pheromone component, (Z)9, (E)11-tetradecadienyl acetate (Z9E11-14:OAc), had arborizations that were restricted to a large glomerulus of the MGC (designated a), situated near the antennal nerve entrance into the antennal lobe. Neurons that were stimulated by (Z)9,(E)12-tetradecadienyl acetate (Z9E12-14:OAc), a second pheromone component, had arborizations in a lateral, smaller glomerulus of the MGC (designated c), while receptor neurons specifically tuned to a behavioural antagonist, (Z)-9-tetradecenol (Z9-14:OH), projected to a medial glomerulus of the MGC (b). In the female, receptor neurons tuned to the major pheromone component projected to a glomerulus situated at the entrance of the antennal nerve. This glomerulus did, however, not have the size or the structure of the male MGC. A second neuron housed in the same sensillum projected its axon to an ordinary glomerulus situated medially in the antennal lobe.

Transmission of Theileria parva to cattle by Rhipicephalus appendiculatus adults fed as nymphae in vitro on infected blood through an artificial membrane.

A technique is described for the efficient feeding of Rhipicephalus appendiculatus nymphae on cattle blood through an artificial membrane bearing tactile and olfactory stimuli. The effect of four anticoagulation methods on the feeding of nymphae was compared and heparinized blood was found to be the most efficacious, followed by defibrinated blood. Blood treated with acid citrate dextrose (ACD) or ethylenediamine tetraacetate (EDTA) inhibited nymphal feeding. Nymphae fed on heparinized and defibrinated blood obtained engorgement weights within the range of ticks fed on mammalian hosts and they subsequently moulted and fed normally as adults and produced viable eggs. Nymphae fed on membranes using either defibrinated or heparinized blood infected with Theileria parva piroplasma developed salivary gland infections as adult ticks and transmitted East Coast fever (ECF) to susceptible cattle. There were indications that T. parva-infected defibrinated blood was not as infective to the feeding nymphae as the infected heparinized blood. When T. parva-infected heparinized blood was used to feed nymphae through membranes in two experiments, it was found that the infections in the resultant adult ticks could be comparable to those of nymphae fed on donor cattle, but were usually lower. The membrane feeding technique will enable the study of factors affecting the tick and T. parva transmission without the complication of host/T. parva interactions and could be useful for both tick maintenance and Theileria parasite isolation and maintenance.

Artificial-membrane feeding of the ixodid tick, Rhipicephalus appendiculatus, to repletion.

An artificial feeding device was constructed with a Baudruche membrane bearing olfactory and tactile stimuli, to induce voluntary attachment and feeding of the brown ear tick, Rhipicephalus appendiculatus. Preparation of the membrane and experimental conditions used are described. Unfed adults of R. appendiculatus, 2-3 weeks after moulting, were confined on the membrane, which was treated with cattle ear-wash and other stimuli. They attached and commenced sucking blood within 12-48 h. Fully engorged ticks started detaching from the membrane on the 6th day. After a 5-6-day incubation period, they started laying eggs which hatched into normal larvae.