Immunocytochemical localization of pheromone-binding protein in moth antennae

Summary Odorant-binding proteins are supposed to play an important role in stimulus transport and/or inactivation in olfactory sense organs. In an attempt to precisely localize pheromone-binding protein in the antenna of moths, post-embedding immunocytochemistry was performed using an antiserum against purified pheromone-binding protein of Antheraea polyphemus. In immunoblots of antennal homogenates, the antiserum reacted exclusively with pheromone-binding protein of A. polyphemus, and cross-reacted with homologous proteins of Bombyx mori and Autographa gamma. On sections of antennae of male A. polyphemus and B. mori, exclusively the pheromone-sensitive sensilla trichodea are labelled; in A. gamma, label is restricted to a subpopulation of morphologically similar sensilla trichodea, which indicates that not all pheromone-sensitive sensilla contain the same type of pheromone-binding protein and accounts for a higher specificity of pheromone-binding protein than hitherto assumed. Within the sensilla trichodea, the extracellular sensillum lymph of the hair lumen and of the sensillum-lymph cavities is heavily labelled. Intracellular label is mainly found in the trichogen and tormogen cells: in endoplasmic reticulum, Golgi apparatus, and a variety of dense granules. Endocytotic pits and vesicles, multivesicular bodies and lysosome-like structures are also labelled and can be observed not only in these cells, but also in the thcogen cell and in the receptor cells. Cell membranes are not labelled except the border between thecogen cell and receptor cell and the autojunction of the thecogen cell. The intracellular distribution of label indicates that pheromone-binding protein is synthesized in the tormogen and trichogen cell along typical pathways of protein secretion, whereas its turnover and decomposition does not appear to be restricted to these cells but may also occur in the thecogen and receptor cells. The immunocytochemical findings are discussed with respect to current concepts of the function of pheromone-binding protein.     

Drosophila “enhancer-trap” transposants: Gene expression in chemosensory and motor pathways and identification of mutants affected in smell and taste ability

We have isolated about a thousandDrosophila P-element transposants that allow thein situ detection of genomic enhancer elements by a histochemical assay for β-galactosidase activity. We summarize the β-galactosidase staining patterns of over 200 such transposants in the adult. Our aim was to identify genes that are likely to be involved in the chemosensory and motor pathways ofDrosophila. Based on β-galactosidase expression patterns in the tissues of our interest, we have chosen some strains for further analysis. Behavioral tests on a subset of the transposants have, in addition, identified several strains defective in their chemosensory responses.     

Temporal analysis of adaptation in moth (Trichoplusia ni) pheromone receptor neurons

Summary The temporal pattern of response in chemoreceptor neurons reflects both the temporal distribution of stimuli and the timing of signal transduction, action potential generation and propagation. Here we analyze the temporal characteristics of the responses elicited in pheromone receptor neurons by computer-controlled rectangular pulses of odorant. Extracellular recordings from the HS sensilla trichodea on the antenna of male Trichoplusia ni reveal the activity of two neurons: the A neuron, which responds to the major component of the female pheromone blend, (Z)7-dodecenyl acetate and the B neuron, which responds to (Z)7-dodecenol. B neurons were divided into two classes (HR, LR), based on the magnitude and temporal pattern of their response to (Z)7-dodecenol. Most A and HR B neurons responded to rectangular pulses of various durations (0.1–40 s) with an initial phasic burst (100 ms), followed by a slowly declining tonic component. At moderate and elevated pheromone doses, prolonged stimulation resulted in significant reductions in the tonic response levels (adaptation); stimuli of increasing duration effected greater adaptation. Most LR B neurons lacked a phasic response component and showed virtually no adaptation with prolonged stimulation. Pheromone receptor neurons may differ in both their spectral and temporal response properties which may provide the animal with additional sensory information for blend discrimination and spatial orientation in complex natural pheromone plumes. The potential functional value of adaptation in the moth pheromone communication system is discussed.Abbreviations Z7,12:AC (Z)7-dodecenyl acetate - Z7,12:OH (Z)7-dodecenol - HR High response - LR Low response - HS High sensitivity     

Odorant response of individual sensilla on theDrosophila antenna

We have documented odor responses of all morphological classes of sensilla on the surface of theDrosophila antenna: sensilla basiconica, sensilla trichodea, and sensilla coeloconica. Both subtypes of s. basiconica, large and small, respond to odors. S. trichodea fall into different functional types. Type 1 appears narrowly tuned, as it responded only tocis-vaccenyl acetate, believed to be a pheromone. Type 2 responded totrans-2-hexenal and 4-methyl cyclohexanol. These two types of s. trichodea are differentially distributed on the antennal surface, and have dramatically different frequencies of spontaneous action potentials. Likewise, there are multiple types of s. coeloconica. One type is broadly tuned, responding most strongly to a test stimulus of butyric acid, but also to a variety of other odors; it is restricted to the dorso-medial portion of the third antennal segment. A second type gave detectable responses only totrans-2-hexenal. These results demonstrate that all classes of sensilla are olfactory, and they reveal the organizational complexity of theDrosophila olfactory system.     

Density-dependent predation by skunks using olfactory search images

The formation of search images can create density-dependent predation. Predators have been shown to form search images when searching for many small prey items in one feeding session. This paper reports experiments that test whether striped skunks can form olfactory search images in other situations: when prey are found over several days, when prey are large, and when prey are found in certain habitats. Striped skunks were raised in captivity, and their reaction distance to food was measured outside in a natural grassy area. In experiment 1 skunks were offered many small food items for several days in a row. From one day to the next, skunks initially detected food from further away, they increased detection distance faster and their maximum detection distance increased – i.e., they formed olfactory search images faster and stronger from one day to the next. In experiment 2 skunks formed search images over several days when finding only one large food item per day. In experiment 3 skunks lost olfactory search images when they entered habitats in which they had previously searched for another type of food. These long-term search images magnify the effects of short-term search images, extend the effects of short- term search image to longer time spans, and affect different species from short-term search images. Received: 26 July 1996 / Accepted: 13 December 1996     

Volatile cues can drive the oviposition behavior in Odonata

Selection for the oviposition site represents the criterion for the behavioral process of habitat selection for the next generation. It is well known that in Odonata the most general cues are detected visually, but laboratory investigations on the coenagrionid Ischnura elegans showed through behavioral and electrophysiological assays that adults were attracted by olfactory cues emitted by prey and that males of the same species are attracted by female odor.The results of the present behavioral and electrophysiological investigations on I. elegans suggest the involvement of antennal olfactory sensilla in oviposition behavior. In particular, I. elegans females laid in the laboratory significantly more eggs in water from larval rearing aquaria than in distilled or tap water. Moreover, the lack of preference between rearing water and tap water with plankton suggests a role of volatiles related to conspecific and plankton presence in the oviposition site choice. I. elegans may rely on food odor for oviposition site selection, thus supporting the predictions of the “mother knows best” theory. These behavioral data are partially supported by electroantennographic responses. These findings confirm a possible role of olfaction in crucial aspects of Odonata biology.     

Perinatal visceral events and brain mechanisms involved in the development of mother-young bonding in sheep

In sheep the onset of maternal responsiveness and the development of the mutual mother-young bond are under the combined influence of hormonal and visceral somatosensory stimulations. These stimuli are provided in the mother by parturition (via steroids and vaginocervical stimulation) and in the neonate by the first suckling episodes (via cholecystokinin and oro-gastro-intestinal stimulation). In addition, each partner relies on specific chemosensory stimulation for reciprocal attraction: amniotic fluids for the mother, colostrum for the young. In the ewe parturition activates several brain structures to respond specifically to sensory cues emanating from the young. The main olfactory bulbs undergo profound neurophysiological changes when exposed to offspring odors at parturition. Additional activations in the hypothalamus - preoptic area - and the amygdala - medial and cortical nuclei - also contribute to maternal responsiveness and memorization of lamb odors. In the neonate, post-ingestive stimulations activate the brain stem via vagal afferents. Like in the ewe, several regions of the hypothalamus and the amygdala respond to colostrum ingestion suggesting common ground for the integrative neural processes involved in early learning and bonding. This leads to rapid visual and auditory recognition in both partners although olfaction remains important in the ewe to display selective nursing. It is concluded that the biological basis for the development of maternal and filial bonding in sheep presents striking similarities.     

Ventromedial hypothalamic nucleus lesions disrupt olfactory mate recognition and receptivity in female ferrets

Previous research showed that ferrets of both sexes rely on the perception of conspecifics' body odors to identify and motivate approach towards opposite-sex mating partners, and exposure to male body odors stimulated Fos expression in an olfactory projection circuit of female, but not male, ferrets that terminates in the ventromedial hypothalamic nucleus (VMH). We asked whether the female-typical preference of ferrets to approach male as opposed to female body odors in Y-maze tests would be disrupted by VMH lesions. Sexually experienced female ferrets were ovo-hysterectomized prior to receiving bilateral electrolytic lesions of the VMH, the preoptic area/anterior hypothalamus (POA/AH) or a sham operation. Subsequently, while receiving estradiol benzoate, females that received either complete or partial bilateral lesions of the VMH approached volatile odors from an anesthetized male on significantly fewer trials than females given POA/AH lesions or a sham operation. Both groups of ferrets with VMH lesion damage reliably discriminated between volatile anal scents as well as urinary odors from the 2 sexes in home cage habituation/dishabituation tests, suggesting that their odor-based sex discrimination remained intact. Females with complete bilateral VMH lesions showed significantly lower acceptance of neck gripping from a stimulus male (receptivity) and more aggression towards the male than all other groups of female subjects. Estrogen-sensitive neurons in the VMH appear to play a central role in female-typical neural processing of odor inputs leading to a preference to seek out a male sex partner, in addition to facilitating females' sexual receptivity.