The molecular basis of odor coding in the Drosophila antenna

We have undertaken a functional analysis of the odorant receptor repertoire in the Drosophila antenna. Each receptor was expressed in a mutant olfactory receptor neuron (ORN) used as a "decoder," and the odor response spectrum conferred by the receptor was determined in vivo by electrophysiological recordings. The spectra of these receptors were then matched to those of defined ORNs to establish a receptor-to-neuron map. In addition to the odor response spectrum, the receptors dictate the signaling mode, i.e., excitation or inhibition, and the response dynamics of the neuron. An individual receptor can mediate both excitatory and inhibitory responses to different odorants in the same cell, suggesting a model of odorant receptor transduction. Receptors vary widely in their breadth of tuning, and odorants vary widely in the number of receptors they activate. Together, these properties provide a molecular basis for odor coding by the receptor repertoire of an olfactory organ.     

Effect of motor stimulation and stretching on afferent activity of the neuromuscular spindle isolated from the frog]

The arrangement of muscle spindles in m. ext. long. dig. IV has been examined by microdissection. It is confirmed that spindle systems generally appear to consist of individual receptors. Stimulation effects of fast motor fibres (conduction velocities greater than 12 m/sec) on the spindles of the same muscle were studied. Receptors were isolated with their nerves and the appropriate spinal roots, the latter ones were used for stimulating efferent fibres and recording sensory discharges. Single shocks to the ventral root filaments caused afferent responses ranging from a single action potential to a train of impulses. During repetitive stimulation (train of stimuli at frequency of 10 to 150/sec) a marked increase in afferent activity was found. Afferent activity could be driven by the frequency of stimuli ("driving") and the stimulus/action potentials ratio varied from 1:1 to 1:3 or more. The rate of sensory discharge depended on the frequency of stimuli: the maximum effect, was attained at 30 to 50 stimuli/sec and, in the most responsive receptors, up to 80 stimuli/sec. Slight increases of the initial lengths of the receptors caused facilitation of sensory responses to motor stimulation. Moreover, impairing effects, which appear during sustained or high-frequency stimulation, possibly related to fatigue in intrafusal neuromuscular transmission, could be relieved by increasing the initial length. The repetitive stimulation of fast fusimotor fibres increased both dynamic and static responses and also raised the afferent activity after a period of stretching, when usually a depression occurs; these effects varied according to the preparation, its initial tension and the frequency of stimulation. The main feature of the examined motor fibres, when stimulated, is the constant excitatory action on muscle spindle static response. Results are discussed. It is suggested that the different characteristics of intrafusal muscle fibres, the receptor initial tension and the frequency of motor units discharges, may together affect muscle spindles static or dynamic performance.     

Electrophysiological evidence for a topographical projection of the nasal mucosa onto the olfactory bulb of the frog

Three olfactory nerve branches respectively subserving either a medial, an intermediate, or a lateral region of the dorsal olfactory receptor sheet of the bullfrog Rana catesbeiana were electrically stimulated with bipolar platinum hook electrodes. Extracellular single unit responses from 93 second-order cells in different regions of the olfactory bulb were recorded with metal-filled glass micropipets. The excitatory responsiveness of each unit to the stimulation of each of the three nerve branches (response profile) was determined. Some units were sensitive to stimulation of each of the three nerve branches, thus suggesting a wide projection from the entire receptor sheet. On the other hand, other units were more selective. Of this latter group, units in the lateral bulb were excited by nerve branches subserving the more lateral regions of the receptor sheet; units in the medial bulb were excited by the nerve branches subserving the more medial regions of the receptor sheet. These data provide electrophysiological evidence for a topographical projection of the olfactory receptor sheet onto the olfactory bulb, and further suggest that the projections onto different bulbar cells vary in degree of localization.     

Odorant receptors activated by amino acids in sensory neurons of the channel catfish Ictalurus punctatus

Odorant receptors activated by amino acids were investigated with patch- clamp techniques in olfactory receptor neurons of the channel catfish, Ictalurus punctatus. The L-isomers of alanine, norvaline, arginine, and glutamate, known to act predominantly on different olfactory receptor sites, activated nondesensitizing inward currents with amplitudes of - 2.5 to -280 pA in olfactory neurons voltage-clamped at membrane potentials of -72 or -82 mV. Different amino acids were shown to induce responses in the same sensory neurons; however, the amplitude and the kinetics of the observed whole cell currents differed among the stimuli and may therefore reflect activation of different amino acid receptor types or combinations of receptor types in these cells. Amino acid- induced currents appeared to have diverse voltage dependence and could also be classified according to the amplitude of the spontaneous channel fluctuations underlying the macroscopic currents. A mean single- channel conductance (gamma) of 360 fS was estimated from small noise whole-cell currents evoked by arginine within the same olfactory neuron in which a mean gamma value of 23.6 pS was estimated from ''large noise'' response to norvaline. Quiescent olfactory neurons fired bursts of action potentials in response to either amino acid stimulation or application of 8-Br-cyclic GMP (100 microM), and voltage-gated channels underlying generation of action potentials were similar in these neurons. However, in whole-cell voltage-clamp, 8-Br-cyclic GMP evoked large rectangular current pulses, and single-channel conductances of 275, 220, and 110 pS were obtained from the discrete current levels. These results suggest that in addition to the cyclic nucleotide-gated transduction channels, olfactory neurons of the channel catfish possess a variety of odor receptors coupled to different types of transduction channels.     

Leishmania amazonensis: chemotaxic and osmotaxic responses in promastigotes and their probable role in development in the phlebotomine gut

Taxic responses may play a role in development of Leishmania in their phlebotomine sand fly vectors. They are possibly responsible for movement of the parasites towards the anterior regions of the gut, from where they would be transmitted to the vertebrate host. A methodology capable to distinguish chemotaxic from osmotaxic responses was described and used to characterise taxic responses in Leishmania promastigotes. These were able to respond to chemotaxic as well as to osmotaxic stimuli. Like bacteria, promastigotes were capable to undergo "adaptation," a phenomenon by which they stop responding to a continuos stimulus. A model capable to explain how a relatively small number of different receptors works to perceive gradients in chemotaxic responses was proposed. According to this model, these receptors possess low specificity and a wide range of affinities varying from high to low. A low specificity makes the same receptor able to bind to a large number of different but structurally related molecules and; a wide range of affinities (considering a population of receptors), implies that the number of receptors "occupied" by attractant molecules along a gradient would go growing step by step.     

Olfactory receptor cell responses of pigeon to some odors

A preparation has been developed in the pigeon which allows recording of the electrical activity from an olfactory nerve twig containing the nonmyelinated axons of a small group of olfactory receptor cells. The pigeon's response to n-amylacetate is vigorous and stable, like that of other air-breathing animals. Responses in the olfactory receptor cells in the pigeon increased in magnitude with increase in the odor concentration. An olfactory nerve twig produced a different magnitude of responses to the various odor stimuli. When an odor stimulation was applied to the olfactory mucosa, the two different olfactory nerve twigs which were separated from the same olfactory nerve bundle produced a different magnitude of responses. The differences may be dependent on several factors.     

Ultraviolet and green receptors in principal eyes of jumping spiders

Spectral sensitivities of cells in principal eyes of the jumping spider Phidippus reqius were measured using techniques of intracellular recording. Three types of cells were found. UV cells had peak sensitivities at 370 nm and were over 4 log units less sensitive at wavelengths longer than 460 nm. Green-sensitive cells had spectral sensitivities which were well fit by nomogram curves peaking at 532 nm. UV-green cells had dual peaks of sensitivity at about 370 and 525 nm, but the ratios of UV-to-green sensitivities varied over a 40: 1 range from cell to cell. Moreover, responses of UV-green cells to flashes of UV light were slower than to flashes of green light. Segregation of receptor types into the known layers of receptors in these eyes could not be shown. It is concluded that jumping spiders have the potential for dichromatic color vision.     

Discrimination between enantiomers of linalool by olfactory receptor neurons in the cabbage moth Mamestra brassicae (L.)

Plants emit complex blends of volatiles, including chiral compounds that might be detected by vertebrates and invertebrates. Insects are ideal model organisms for studying the underlying receptor neuron mechanisms involved in olfactory discrimination of enantiomers. In the present study, we have employed two-column gas chromatography linked to recordings from single olfactory receptor neurons of Mamestra brassicae, in which separation of volatiles in a polar and a chiral column was performed. We here present the response properties of olfactory receptor neurons tuned to linalool. The narrow tuning of these receptor neurons was demonstrated by their strong responses to (R)-(-)-linalool, the weaker responses to the (+)-enantiomer as well as a few structurally related compounds, and no responses to the other numerous plant released volatiles. The enantioselectivity was verified by parallel dose-response curves, that of (R)-(-)-linalool shifted 1 log unit to the left of the (S)-(+)-linalool curve. A complete overlap of the temporal response pattern was found when comparing the responses of the same strength. Analysis of the spike amplitude and waveform indicated that the responses to the two enantiomers originated from the same neuron.     

Dose-response characteristics of glomerular activity in the moth antennal lobe

Odours are represented as unique combinations of activated glomeruli in the antennal lobes of insects. Receptor neurons arborizing in the glomeruli are not only qualitatively selective, but in addition respond to variations in stimulus concentration. As each glomerulus likely represents a single receptor neuron type, optical recordings of calcium changes in insect antennal lobes show how concentration variations affect a large population of afferents. We measured the glomerular responses in the moth Spodoptera littoralis to different concentrations of plant-related odorants. Localized calcium responses were shown to correspond to individual glomeruli. We found that the dynamic range of glomerular responses spanned 3-4 log units of concentration and the most strongly responding glomeruli often reached a plateau at high stimulus doses. Further, we showed that the single most active glomerulus was often not the same across concentrations. However, if the principal glomerulus moved, it was generally to an adjacent or proximal glomerulus. As concentration increased, a higher number of glomeruli became activated. Correlations of glomerular representations of the same compound at different doses decreased as the difference in concentration increased. Moreover, representations evoked by different odorants were more correlated at high than at low doses, which means that the uniqueness of activity patterns decreased with increasing concentration. Thus, if odours are coded as spatial patterns of glomerular activity, as has been suggested, these olfactory codes are not persistent across concentrations.     

Temporal patterns and selectivity in the unitary responses of olfactory receptors in the tiger salamander to odor stimulation

Temporal patterns and selectivity in unitary responses of 100 single olfactory receptors in the tiger salamander to odor stimulation were investigated. An olfactometer which permitted control of stimulus concentration, duration, and flow rate was calibrated with a gas chromatograph. Stimulus pulses were monitored by recording the electroolfactogram from the surface of the olfactory epithelium. Both diphasic and triphasic spikes were recorded extracellularly. No discernible differences in types of responses, reproducibility of responses, and cross-unit distribution of spontaneous rates distinguished diphasic from triphasic units. The cross-unit selectivity in responses to the seven olfactory stimulants used and the range of odorant concentrations which effectively evoked these responses suggest variations in types and number of types of receptive sites on each cell. Temporal patterns in the unitary responses were generally less complex than those observed in the olfactory bulb. Phasic stimulations evoked phasic patterns. Tonic stimulations evoked phasic/tonic patterns. Occasionally poststimulus depressions or elevations in firing rates were observed. The nature of these patterns varied somewhat with odorant concentration for a particular unit.     

A computational system for simulating and analyzing arrays of biological and artificial chemical sensors

We have designed an approach for modeling olfactory pathways by which one can explore how the properties of individual receptors affect the information coding capacity of an entire system. The effect of receptor tuning breadth on system performance was explored explicitly. We presented model sensory arrays with sets of stimuli randomly and uniformly distributed in an "olfactory space". Arrays of uniformly sized model receptors responding to 25-35% of the stimuli gave the best performance as measured by the ability to capture the most information about the stimulus set. Arrays of variably sized model receptors that were both more broadly and more narrowly tuned than this optimum could, however, perform better than uniform arrays. This method and the results obtained using it suggest a framework for considering the growing body of evidence on the functional properties of individual olfactory receptor and relay neurons from a systems coding perspective.     

数种蜱类哈氏器感器的电生理反应

本实验用电生理方法研究了中华革蜱(Dermacentor sinicus)、森林革蜱(Derraacentor silvarum),和日本血蜱(Haemaphysalis japonica)成虫对不同化学刺激物(氨、冰醋酸、二氧化碳)和性信息素的成分(2,6-二氯酚、对-甲酚、苯酚和水杨醛),以及对温度变化和机械刺激的反应.从哈氏器感觉毛基部记录到的神经传人冲动用信号处理机进行脉冲密度分析,结果表明:(1)哈氏器上不同部位的感觉毛对各种刺激的敏感性不同;(2)蜱的嗅觉反应神经传入冲动有连续性和间歇性两种类型,并且有慢适宜的特点;(3)性信息素的成分中,2,6-二氯酚刺激引起的反应最强,是它的主要成分;(4)在一定浓度范围内,随着刺激物浓度加大反应增强;(5)吸血五天以后的雄蜱对氨的反应减弱,而对2,6-二氨酚的反成增强,吸血七天后反应最强;(6)两种革蜱对各种刺激反应的区别不大.     

Processing of vibratory and acoustic signals by ventral cord neurones in the cricket Gryllus campestris

The responses of single vibratory receptors and ascending ventral cord interneurones were studied extracellularly in Gryllus campestris L. The physiology of the vibration receptors resembled those found in tettigoniids and locusts. The frequency responses of the subgenual receptors provide two possible cues for central frequency discrimination: differences in mean tuning between groups of receptors in the different leg pairs and a range of receptors tuned to different frequencies within one subgenual organ.Most of the ascending vibratory interneurones were highly sensitive in either the low or high frequency range. Broadbanded neurones were less sensitive. The characteristic sensitivity peaks of these units are due mainly to receptor inputs from a particular leg pair, although most central neurones receive inputs from all 6 legs. Only one neurone type, TN1 received excitatory inputs from both auditory and vibratory receptors; its responses were greatly enhanced by the simultaneous presentation of both stimulus modes. The responses to sound stimuli of AN2, on the other hand, were inhibited by vibration. No other auditory interneurones investigated were influenced by inputs from vibration receptors. Central processing of vibratory information in the cricket is compared with that of tettigoniids and locusts.     

Rezeptorpotential und Nervenimpulse einzelner olfaktorischer Sensillen der Insektenantenne

Summary Using glass-capillary microelectrodes, receptor potentials and nerve impulses were recorded from single olfactory units on the antennae of some coleoptera and lepidoptera. These units proved to be the receptor cell endings of sensilla basiconica. Qualitative as well as quantitative differences of responses from single olfactory units might be sufficient for the central-nervous-system to discriminate a rather large number of different odors.     

Hyperpolarizing receptor potentials in lobster olfactory receptor cells: implications for transduction and mixture suppression

Physiological studies of olfactory receptor cells have focusedon excitatory responses, in part because the evidence for inhibitoryresponses from extracellular recordings, although long-standing,has been equivocal. Intracellular recording from the olfactorycells of two species of lobsters revealed that small but concentrationdependentand repeatable hyperpolarizing receptor potentials could beevoked by a mixture of L-arginine, L-cysteine and L-proline,as well as by histamine. Large, depolarizing receptor potentialswere evoked in the same cells by a complex odor mixture. Simultaneousapplication of depolarizing and hyperpolarizing stimuli reducedthe magnitude of the evoked depolarization. These results implythat multiple, opposing transduction mechanisms are presentin single lobster olfactory receptor cells and reveal a noncompetitivemechanism for peripheral mixture suppression.     

Stimulus-response properties of cricket cereal filiform receptors

The dynamic ranges and stimulus-response properties of a large sample of cereal filiform receptors in Acheta domesticus were investigated electrophysiologically. The relation between receptor response and stimulus velocity was a sigmoid function, the log-linear portion of which spanned 1–1.5 log units of peak air-current velocity. Different receptors responded over different but overlapping velocity ranges, such that the system velocity sensitivity range spanned at least 2.5 log units. Plots of receptor response amplitude vs. stimulus direction were sinusoidal, with a period of 360°. Long-hair receptors responded in phase with air-current velocity, and intermediate-hair receptors responded in phase with air-current acceleration. These results extend those of Shimozawa and Kanou (1984a) and Kämper and Kleindienst (1990), in which the dynamics of receptor responses were shown to depend on hair length. When individual hairs were directly mechanically deflected, their receptors responded in phase with the first derivative of hair deflection. The signal transform between the air-current stimulus and the receptor response is comprised of two processes, one biomechanical/aerodynamic and one membrane biophysical. The results of this study suggest that the parametric sensitivities of receptors are primarily determined by hair biomechanical/aerodynamic properties.Abbreviation IR infrared     

The response properties of retinula cells in the flyCalliphora erythrocephala as a function of the wavelength and polarization properties of visible and ultraviolet light

Detailed functional response properties of the class (1–6) retinula cells in the insectCalliphora erythrocephela have been determined as a function of the wavelength of monochromatic light and polarization conditions over a six log unit range of light adaptation levels. These stimuli were produced as a small parallel beam that could combine various wavelengths in both the ultraviolet and visible regions whose polarization planes could be individually controlled. The wild type and white-eyed mutant were studied and shown to have no significant differences in their response properties. It was shown that previous tests using transient stimuli, large compared to the adaptation levels produced responses of high orders of nonlinearity and rapidly changing transient properties. In contrast, the use of white-noise tests did not cause changes in the basic cell properties as established by a given light adaptation condition. These tests accurately model the visual conditions of a wide and representative range of normal behavioral conditions and the model response data thus obtained can be directly correlated to important physiological tests for the same stabilized adaptation conditions. The simultaneous application of both visible and ultraviolet monochromatic light revealed a complex response interaction which changed the self response contribution of the ultraviolet light from that due only to ultraviolet light but had no effect on the visible light response. It also produced a mutual interaction. This effect was found to be the same for all members of the (1–6) class of cells when the light was unpolarized but varied for different members of the class for polarized light depending upon the relative angles of theE-vectors for the two types of light which in turn vary for different members of the six cell classes.     

Decoding complex chemical mixtures with a physical model of a sensor array

Combinatorial sensor arrays, such as the olfactory system, can detect a large number of analytes using a relatively small number of receptors. However, the complex pattern of receptor responses to even a single analyte, coupled with the non-linearity of responses to mixtures of analytes, makes quantitative prediction of compound concentrations in a mixture a challenging task. Here we develop a physical model that explicitly takes receptor-ligand interactions into account, and apply it to infer concentrations of highly related sugar nucleotides from the output of four engineered G-protein-coupled receptors. We also derive design principles that enable accurate mixture discrimination with cross-specific sensor arrays. The optimal sensor parameters exhibit relatively weak dependence on component concentrations, making a single designed array useful for analyzing a sizable range of mixtures. The maximum number of mixture components that can be successfully discriminated is twice the number of sensors in the array. Finally, antagonistic receptor responses, well-known to play an important role in natural olfactory systems, prove to be essential for the accurate prediction of component concentrations.     

Peripheral regulation by ecdysteroids of olfactory responsiveness in male Egyptian cotton leaf worms, Spodoptera littoralis

Physiological and behavioral plasticity allows animals to adapt to changes in external (environmental) and internal (physiological) factors. In insects, the physiological state modulates adult behavior in response to different odorant stimuli. Hormones have the potential to play a major role in the plasticity of the olfactory responses. To explore if peripheral olfactory processing could be regulated by steroid hormones, we characterized the molecular, electrophysiological, and behavioral response to changes in endogenous hormone levels in adult male Spodoptera littoralis. The expression of the receptor complex (EcR/USP) was localized by in situ hybridization in the olfactory sensilla of antennae. Injections of 20-hydroxyecdysone (20E) induced an ecdysteroid signaling pathway in antennae and increased expression of the nuclear receptors EcR, USP and E75. Diacylglycerol kinase (DGK) and CaM expression were also up-regulated by 20E. Taken together, these molecular, electrophysiological, and behavioral results suggest a hormonal regulation of the peripheral olfactory processing in S. littoralis.     

Random expression of main and vomeronasal olfactory receptor genes in immature and mature olfactory epithelia of Fugu rubripes

Main olfactory receptor genes were isolated from a seawater fish, Fugu rubripes (pufferfish), and characterized. Two subfamilies of genes encoding seven transmembrane receptors were identified; one consists of five or more members, termed FOR1-1 to 5 of FOR1 subfamily, and the other appears to be a single copy gene, termed the FOR2 subfamily. FOR1 members show extremely high amino acid sequence similarities of about 95% to one another, and are distantly related to catfish-1 with the highest similarity of 37%. FOR2 shows 43% similarity to goldfish-A28. Phylogenically, both FOR members are categorized among pedigrees of the fish main olfactory receptor family outside the mammalian receptor family, although similarities between Fugu receptors and those of fresh-water fishes are lower than those among fresh-water fishes. In situ hybridization shows that both subfamilies of receptor genes are expressed randomly over the olfactory epithelium throughout all developmental stages, and no segregation of the signals was found. On the other hand, when three members of a vomeronasal olfactory receptor gene family, related to the Ca(2+)-sensing receptor, were used as probes, they were also randomly expressed over the same epithelium as the main olfactory receptors. This is in contrast to the expression profiles observed for zebrafish and goldfish, where the main or vomeronasal olfactory receptors are expressed in segregated patterns. It is thus suggested that the expression pattern of fish olfactory receptors varies depending on the species, although fish olfactory receptors are highly related to one another in their primary structures, and are phylogenically distinct from those of mammals.