Pre-exposure affects the olfactory response of Drosophila melanogaster to menthol

A modification of the trap assay (Woodard et al., 1989) was used to evaluate the response of Drosophila melanogaster (Meigen) to food media containing menthol. Dose-response curves for flies to mentholic foods were produced for flies that had been pre-exposed to menthol, during development and adult life, and flies that had not been exposed to menthol before the assay. Mentholic food media were less attractive to Drosophila than plain food medium. Rearing flies on a medium containing menthol reduced their aversion to some concentrations of menthol. The rearing effect was not simply due to lowered general activity levels resulting from developing in a medium containing menthol. There was a threshold concentration of menthol in the rearing medium below which we found no induced behavioural change.     

Lactosamine differentially affects olfactory sensory neuron projections to the olfactory bulb

During embryonic development, olfactory sensory neurons extend axons that form synapses with the dendrites of projection neurons in glomeruli of the olfactory bulb (OB). The glycosyltransferase beta3GnT1 regulates the expression of 1B2-reactive lactosamine glycans that are mosaically distributed among glomeruli. In newborn beta3GnT1-/- mice, lactosamine expression is lost, and many glomeruli fail to form. To determine the role of lactosamine in OB targeting, we analyzed the trajectories of specific OR axon populations and their reactivity with 1B2 in beta3GnT1-/- mice. mI7 axons and P2 axons, both of which are weakly 1B2+ in wild-type mice, fail to grow to their normal positions in the glomerular layer during early postnatal development and never recover in adult mutant mice. In contrast, many M72 axons, which are always lactosamine negative in wild-type mice, survive but are misguided to the extreme anterior OB in neonatal mutant mice and persist as heterotypic glomeruli, even in adult null mice. These results show that the loss of lactosamine differentially affects each OR population. Those that lose their normal expression of lactosamine fail to form stable connections with mitral and tufted cells in the OB, disappear during early postnatal development, and do not recover in adults. Neurons that are normally lactosamine negative, survive early postnatal degeneration in beta3GnT1-/- mice but extend axons that converge on inappropriate targets in the mutant OB.     

Atlas of olfactory organs of Drosophila melanogaster 2. Internal organization and cellular architecture of olfactory sensilla

Antennae and maxillary palps of Drosophila melanogaster were studied with the electron microscope on serial sections of cryofixed specimens. The number of epidermal cells roughly equals the number of sensilla, except for regions where the latter are scarce or absent. Each epidermal cell forms about two non-innervated spinules, a prominent subcuticular space and a conspicuous basal labyrinth, suggesting a high rate of fluid transport through the sensory epithelium. The internal organization and fine structure of trichoid, intermediate and basiconic sensilla is very similar. Receptor cell somata are invested by thin glial sheaths extending distad to the inner dendritic segments. Further distally, the thecogen cell forms a sleeve around the dendrites, but an extracellular dendrite sheath is absent. At the base of the cuticular apparatus, the inner sensillum-lymph space around the ciliary and outer dendritic segments is confluent with the large outer sensillum-lymph space formed by the trichogen and tormogen cells. All three auxiliary cells exhibit many features of secretory and transport cells but extend only thin basal processes towards the haemolymph sinus. The bauplan and fine structure of coeloconic sensilla differs in the following aspects: (1) the ciliary segment of the dendrites is located deeper below the base of the cuticular apparatus than in the other sensillum types; (2) a prominent dendrite sheath is always present, separating inner and outer sensillum-lymph spaces completely; (3) the apical microlamellae of the auxiliary cells are more elaborate, but free sensillum-lymph spaces are almost absent; (4) there are always four not three auxiliary cells. Morphometric data are presented on the diameter of inner and outer dendritic segments and on the size of receptor cells, as well as of the receptor and auxiliary cell nuclei. The special fine structural features of Drosophila olfactory sensilla are discussed under the aspects of sensillar function and the localization of proteins relevant for stimulus transduction.     

Drosophila olfactory memory: single genes to complex neural circuits

A central goal of neuroscience is to understand how neural circuits encode memory and guide behaviour. Studying simple, genetically tractable organisms, such as Drosophila melanogaster, can illuminate principles of neural circuit organization and function. Early genetic dissection of D. melanogaster olfactory memory focused on individual genes and molecules. These molecular tags subsequently revealed key neural circuits for memory. Recent advances in genetic technology have allowed us to manipulate and observe activity in these circuits, and even individual neurons, in live animals. The studies have transformed D. melanogaster from a useful organism for gene discovery to an ideal model to understand neural circuit function in memory.     

DSL-Notch signaling in the Drosophila brain in response to olfactory stimulation

Neurexin and neuroligin, which undergo heterophilic interactions with each other at the synapse, are mutated in some patients with autism spectrum disorder, a set of disorders characterized by deficits in social and emotional learning. We have explored the role of neurexin and neuroligin at sensory-to-motor neuron synapses of the gill-withdrawal reflex in Aplysia, which undergoes sensitization, a simple form of learned fear. We find that depleting neurexin in the presynaptic sensory neuron or neuroligin in the postsynaptic motor neuron abolishes both long-term facilitation and the associated presynaptic growth induced by repeated pulses of serotonin. Moreover, introduction into the motor neuron of the R451C mutation of neuroligin-3 linked to autism spectrum disorder blocks both intermediate-term and long-term facilitation. Our results suggest that activity-dependent regulation of the neurexin-neuroligin interaction may govern transsynaptic signaling required for the storage of long-term memory, including emotional memory that may be impaired in autism spectrum disorder.     

Extracellular recording of spatiotemporal patterning in response to odors in the olfactory epithelium by microelectrode arrays

In olfactory biosensors, microelectronic sensor chips combined with biological olfactory cells are a promising platform for odor detection. In our investigation, olfactory epithelium stripped from rat was fixed on the surface of microelectrode arrays (MEAs). Electrophysiological activities of olfactory receptor neurons in intact epithelium were measured in the form of extracellular potentials. Based on multi-channel recording performance of MEA and structural and functional integrality of native olfactory epithelium, the spatiotemporal analysis was carried out to study the extracellular activity pattern of neurons in the tissue. The variation of spatiotemporal patterns corresponding to different odors displayed the signals firing image characteristic intuitionally. It is an effective method in the form of patterns for monitoring the state of tissue both in time and space domain, promoting the platform for olfactory sensing mechanism research.     

Inactivation of c-Cbl or Cbl-b differentially affects signaling from the high affinity IgE receptor

The Cbl family of proteins negatively regulate signaling from tyrosine kinase-coupled receptors. Among the three members of this family, only c-Cbl and Cbl-b are expressed in hemopoietic cells. To examine the role of c-Cbl and Cbl-b in Fc epsilon RI signaling, mast cell cultures from wild-type, c-Cbl(-/-), and Cbl-b(-/-) mice were generated. Cell growth rates and cell surface expression of Fc epsilon RI were similar in the different cell populations. Compared with control cells, Cbl-b inactivation resulted in increases in Fc epsilon RI-induced Ca(2+) response and histamine release. Fc epsilon RI-induced tyrosine phosphorylation of total cellular proteins, Syk, and phospholipase C-gamma was also enhanced by Cbl-b deficiency, whereas receptor-initiated phosphorylation of Vav, JNK, and p38 kinases was not changed in these cells. In contrast to Cbl-b, c-Cbl deficiency had no detectable effect on Fc epsilon RI-induced histamine release or on the phosphorylation of total cellular proteins or Syk. The absence of c-Cbl increased the phosphorylation of ERK after receptor stimulation, but resulted in slightly reduced p38 phosphorylation and Ca(2+) response. These results suggest that Cbl-b and c-Cbl have divergent effects on Fc epsilon RI signal transduction and that Cbl-b, but not c-Cbl, functions as a negative regulator of Fc epsilon RI-induced degranulation.     

Role of the olfactory systems and importance of learning in the ewes' response to rams or their odors

In sheep, exposure of seasonally anestrous females to the male or its fleece results in activation of luteinizing hormone (LH) secretion and synchronized ovulation. The study of the neural pathways involved in this phenomenon, commonly named "male effect", show that the main olfactory system plays a critical role in the detection and the integration of the male odor. The accessory olfactory system participates in the perception of the ram odor but does not seem necessary for the endocrine response. According to the hypothesis that the neuroanatomical differences between the two olfactory systems could be associated with different functional roles, we investigated the importance of sexual experience and learning processes in the male effect. Our results showed that female responses depend on previous sexual experience. We also demonstrated that the LH response to male odor could result from an associative learning process. The aim of the present report was to summarize our current knowledge concerning the "male effect" and in particular to clarify the role of sexual experience and learning in the processes involved in this effect.     

Responsiveness of the olfactory receptor cells in dog to some odors

A preparation has been developed in the dog which allows recording the electrical activity from an olfactory nerve twig containing the axons of a small group of olfactory receptor cells. The dog's response to n-pentyl acetate is vigorous and stable, like that of other air-breathing animals. The dog's response magnitude dependence on the nasal flow rate was noticeable for n-pentyl acetates, but not so great as for n-butyric acid. The response to n-butyric acid strongly depends on the nasal flow. The start of the nasal air flow caused an increase of neural activity, which is called flow response. The results show that the nasal flow rate is a very important factor which determines the response to odors. Methyl p-hydroxybenzoate is known as a dog's pheromone, however, this odor caused the feeble response in the electrical activity of the dog's olfactory receptor cells. The differences may be dependent on several factors.     

Perception of odors by a nonlinear model of the olfactory bulb

The behavior of the olfactory bulb is modeled as a network of interconnected cells with nonlinear dynamics. External inputs from sensory neurons are introduced as perturbations to subsets of cells within the network. We describe the attractors of the system and show how they can be classified and ordered according to their varying degrees of symmetry. By studying networks of attractors in the system's phase space, it is shown how different perturbations may evoke specific switches between various patterns of behavior. This ensures that different odors, even if present at extremely low concentrations, are able to evoke a specific spatio-temporal behavior in the olfactory bulb, permitting their unique perception. The model incorporates many of the processes proposed to mediate perception, such as the topographic organisation of sensory systems, destabilization of cortex by sensory input and synchronisation between neurons. It is also consistent with the character of the olfactory electroencephalogram.     

A large family of divergent Drosophila odorant-binding proteins expressed in gustatory and olfactory sensilla.

We identified a large family of putative odorant-binding protein (OBP) genes in the genome of Drosophila melanogaster. Some of these genes are present in large clusters in the genome. Most members are expressed in various taste organs, including gustatory sensilla in the labellum, the pharyngeal labral sense organ, dorsal and ventral cibarial organs, as well as taste bristles located on the wings and tarsi. Some of the gustatory OBPs are expressed exclusively in taste organs, but most are expressed in both olfactory and gustatory sensilla. Multiple binding proteins can be coexpressed in the same gustatory sensillum. Cells in the tarsi that express OBPs are required for normal chemosensation mediated through the leg, as ablation of these cells dramatically reduces the sensitivity of the proboscis extension reflex to sucrose. Finally, we show that OBP genes expressed in the pharyngeal taste sensilla are still expressed in the poxneuro genetic background while OBPs expressed in the labellum are not. These findings support a broad role for members of the OBP family in gustation and olfaction and suggest that poxneuro is required for cell fate determination of labellar but not pharyngeal taste organs.     

Central projections of olfactory receptor neurons from single antennal and palpal sensilla in mosquitoes

In insects, olfactory receptor neurons (ORNs) are located in cuticular sensilla, that are present on the antennae and on the maxillary palps. Their axons project into spherical neuropil, the glomeruli, which are characteristic structures in the primary olfactory center throughout the animal kingdom. ORNs in insects often respond specifically to single odor compounds. The projection patterns of these neurons within the primary olfactory center, the antennal lobe, are, however, largely unknown.We developed a method to stain central projections of intact receptor neurons known to respond to host odor compounds in the malaria mosquito, Anopheles gambiae. Terminal arborizations from ORNs from antennal sensilla had only a few branches apparently restricted to a single glomerulus. Axonal arborizations of the different neurons originating from the same sensillum did not overlap.ORNs originating from maxillary palp sensilla all projected into a dorso-medial area in both the ipsi- and contralateral antennal lobe, which received in no case axon terminals from antennal receptor neurons. Staining of maxillary palp receptor neurons in a second mosquito species (Aedes aegypti) revealed unilateral arborizations in an area at a similar position as in An. gambiae.     

Atlas of olfactory organs of Drosophila melanogaster: 1. Types,external organization,innervation and distribution of olfactory sensilla

The third antennal segment (funiculus) and the maxillary palp are the main and accessory olfactory sense organs of Drosophila melanogaster. Cryofixed antennae and palps revealed a superior preservation of the sensory dendrites and other cellular details as compared to conventional chemical fixation. Extensive cross-section series through funiculus and palp were studied in order to obtain as complete an evaluation as possible of the sensillar complement on these appendages. About 75% of all sensilla on the male and female funiculus were individually studied and their position on the antennal surface mapped. Dimensions of the cuticular apparatus of the various types of sensilla are provided as well as the number of innervating receptor neurons. Particular attention has been paid to the cuticular pores important for stimulus transport and to the sensory dendrites. On the funiculus surface, all sensilla have multiple wall pores: sensilla (s.) trichodea and s. basiconica are single-walled, s. coeloconica are double-walled. The distribution of s. trichodea and s. basiconica follows opposing gradients along a diagonal axis parallel to the axis of the arista from proximo-medial to disto-lateral. In this disto-lateral direction the density of s. trichodea increases while that of the s. basiconica decreases. S. trichodea occur in three subtypes with one, two or three receptor neurons. Basiconic sensilla can be subdivided into three subtypes of large s. basiconica (with two or four receptor neurons), three subtypes of thin s. basiconica (with mostly two, rarely four neurons), and one subtype of small s. basiconica with two receptor neurons. Large s. basiconica occur only in the most proximal region (the ‘LB-zone’); thin s. basiconica are most common in a belt that borders the LB-zone distally, while small s. basiconica are most numerous even further distally along the mentioned diagonal axis in between the s. trichodea. S. intermedia are single-walled, multiporous sensilla which combine features of s. trichodea and s. basiconica; they are found in two subtypes with two or three receptor neurons, in the same region where s. trichodea abound. The s. coeloconica are irregularly distributed over the funicular surface, and occur in two subtypes with two or three receptor neurons. Sexual dimorphism on the antenna is moderate, the female funiculus is a bit longer and carries a larger number of small s. basiconica and large s. basiconica of the LB-I subtype; the male funiculus, however, has more s. trichodea than the female. On the maxillary palp, besides mechanoreceptive s. chaetica, there are only s. basiconica with two receptor neurons. According to the fine structure of their sensory dendrites, three subtypes can be discriminated which are distributed in a random pattern. The functional significance of the described structures and distribution awaits future immunocytochemical and electrophysiological experiments.     

Morphological and ultrastructural characterization of olfactory sensilla in Drosophila suzukii: Scanning and transmission electron microscopy

Drosophila suzukii is a serious horticultural and quarantine pest, damaging various berry crops. Although the active use of olfactory communication in D. suzukii is well-known, their olfactory sensory system has not been comprehensively reported. Therefore, the present study was carried out to understand the morphology, distribution and ultrastructure of olfactory sensilla present in the antennae and maxillary palps of D. suzukii, through scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The olfactory sensilla on the antennae of D. suzukii in both sexes could be classified into three major morphological types, basiconic, trichoid and coeloconic sensilla, according to their shapes. The antennal basiconic sensilla were further divided into three subtypes and the antennal trichoid sensilla into two subtypes, respectively, according to the size of individual sensillum. In contrast to the antennal olfactory sensilla showing diverse morphology, basiconic sensilla was the only type of olfactory sensilla in the maxillary palps of D. suzukii. The basiconic sensilla in the maxillary palps could be further classified into three subtypes, based on their size. Our SEM and TEM observations indicated that multiple nanoscale pores are present on the surface of all types of olfactory sensilla in the antennae and maxillary palps, except coeloconic sensilla. The difference in the morphological types and the distribution of olfactory sensilla suggests that their olfactory functions are different between antennae and maxillary palps in D. suzukii. The results of this study provide useful information for further studies to determine the function of olfactory sensilla in D. suzukii and to understand their chemical communication system.     

A single exposure to novelty differentially affects the accumbal dopaminergic system of apomorphine-susceptible and apomorphine-unsusceptible rats

Individual differences in responses to mild, acute stressors in laboratory animals have commonly been observed in behavioural tests and at the level of hypothalamic-pituitary-adrenal axis responses. These differences are associated with dopamine transmission in the nucleus accumbens. Although the effect of mild stressors on dopamine transmission has been studied with microdialysis, it has not been studied at the level of the catecholaminergic network in the nucleus accumbens. In this study we have used microdialysis to measure extracellular concentrations of dopamine in vivo and immunocytochemistry for the enzyme tyrosine hydroxylase to assess the effect of a single exposure to novelty on the neurochemistry of the nucleus acc umbens in apomorphine-susceptible and apomorphine-unsusceptible rats. These rats are a valid animal model for studying individual differences in responses to environmental stressors and drugs of abuse. We demonstrated that a mild stressor like novelty increased the extracellular concentration of dopamine in the nucleus accumbens in apomorphine-susceptible rats to a larger and longer-lasting degree than in apomorphine-unsusceptible rats. Furthermore we demonstrated that novelty increased the tyrosine hydroxylase-immunoreactive fibre network in the nucleus accumbens shell of apomorphine-susceptible rats, which are rats that are particularly reactive to stressors, but not in the shell of apomorphine-unsusceptible rats, which are rats that are relatively stress-resistant. In conclusion, we have shown that the accumbal dopaminergic system of apomorphine-susceptible rats is more sensitive to an environmental stressor than that of apomorphine-unsusceptible rats. Combined with the fact that these animals also differ in their sensitivity to drugs of abuse, which are known to affect the dopaminergic system, these data provide a solid basis for further studying the differences in the dopaminergic responsiveness to drugs of abuse between apomorphine-susceptible and apomorphine-unsusceptible rats.     

Responses of olfactory receptor neurons in Utetheisa ornatrix to gender-specific odors

We recorded the electrophysiological responses of individual olfactory receptor neurons in sensilla trichodea on the antennae of adult arctiid moths, Utetheisa ornatrix, to stimulation with volatiles associated with both sexes. All trichoid sensilla contain at least two receptor neurons, each with distinct action potential amplitudes and waveforms, that respond dichotomously to male and female odors. Although, neither female neuron responds to extracts of coremata or the male-produced pheromone hydroxydanaidal, they do respond in a gender-specific manner to the volatiles emanating from whole pupae, hemolymph, thoracic froth, and adult animals of several ages. Thoracic froth, which contains pyrrolizidine alkaloids, is thought to play a role in defense. Froth from moths reared on diets, with or without added pyrrolizidine alkaloids, were equally effective in eliciting gender-specific patterns of response. Male trichoid receptor neurons respond to these same materials with similar patterns of activation. These receptor neurons provide information about substances, which we have termed “gender odors,” that are persistently emitted by nearby animals. These substances do not appear to be the same as those already known to be involved in defense or the sexual dialog between individuals. Accepted: 16 March 2000     

Genetic and developmental factors in the olfactory response of Drosophila melanogaster larvae to alcohols.

Olfactory responses of Drosophila melanogaster larvae to a homologous series of primary alcohols (methanol ... decanol) were tested. Alcohols at either extreme of the chain lengths studied (methanol, ethanol and decanol) evoked no significant responses. Heptanol and nonanol both produced dose-independent responses, larvae being attracted to heptanol and repulsed by nonanol. The remaining alcohols elicited dose-related attractive responses. Responses to hexanol and nonanol decline with increasing larval age. Genetic differences were found for the response to heptanol, with larvae from a Japanese strain, Katsunuma, being indifferent to this substance. Chromosome exchange revealed that a major factor involved in the response to heptanol is located on chromosome II; factors on chromosome III quantitatively modulate this response. Three mutant strains were isolated following EMS mutagenesis of chromosome III. These three strains, IndifferentA, IndifferentB and IndifferentC, show incomplete or total anosmia when stimulated with nonanol. Adult flies from these strains show similar effects. IndifferenB and C strains are dominant over the Canton-S control strain; the IndifferentA strain shows semi-dominance. Results are discussed in the light of the ecology of Drosophila larvae and the relation between olfactory stimulus and receptor conformation and number.