Olfactory discrimination ability for homologous series of aliphatic alcohols and aldehydes.

We tested the ability of human subjects to distinguish between members of homologous series of aliphatic alcohols (ethanol to n-octanol) and aldehydes (n-butanal to n-decanal). In a forced-choice triangular test procedure 20 subjects per series were repeatedly presented with all 21 binary combinations of the seven stimuli and asked to identify the bottle containing the odd stimulus. We found (i) that as a group, the subjects performed significantly above chance level in all tasks but two with the alcohols and all tasks but four with the aldehydes, and thus were clearly able to discriminate between most of the odor pairs presented; (ii) marked interindividual differences in discrimination performance, ranging from subjects who were able to significantly distinguish between all 21 odor pairs of a series to subjects who failed to do so with the majority of tasks; and (iii) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length for both homologous series. This suggests that carbon chain length may be one of presumably several determinants of the interaction between stimulus molecule and receptor, and thus may be a molecular property affecting odor quality of aliphatic alcohols and aldehydes.     

Olfactory discrimination of aliphatic odorants at 1 ppm: too easy for CD-1 mice to show odor structure–activity relationships?

Using an operant conditioning paradigm we tested the ability of CD-1 mice to discriminate between 25 odorants comprising members of five homologous series of aliphatic odorants (C4-C8) presented at a gas phase concentration of 1 ppm. We found (a) that all mice significantly discriminated between all 50 stimulus pairs that involved odorants sharing the same functional group, but differing in carbon chain length, as well as between all 50 stimulus pairs that involved odorants sharing the same carbon chain length but differing in functional group, (b) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with the acetic esters and the 2-ketones, but not with the 1-alcohols, n-aldehydes, and n-carboxylic acids tested, and (c) that odorant pairs differing in functional group were significantly better discriminated than odorant pairs differing in carbon chain length. These findings demonstrate that CD-1 mice have excellent discrimination ability for structurally related aliphatic odorants, that correlations between discrimination performance and structural similarity of odorants are odorant class-specific rather than a general phenomenon, and that both carbon chain length and type of functional group play an important role for odor quality coding in mice.     

Detection thresholds for phenyl ethyl alcohol using serial dilutions in different solvents

Detection thresholds are typically obtained by presenting a subject with serial dilutions of an odorant. Many factors, including the solvent used to dilute the odorant, can influence the measurement of detection thresholds. Differences have been reported in detection thresholds for phenyl ethyl alcohol (PEA) when different solvents are used. In this study we used gas chromatography (GC) to investigate further the effect of solvent on odor detection thresholds. We used a single ascending method and serial dilutions of PEA in four different solvents--liquid paraffin (LP), mineral oil (MO), propylene glycol (PG) and dipropylene glycol (DPG)--to determine the PEA thresholds for 31 adult subjects. For each solvent, we prepared eight serial log base 10 step dilutions (1-8), with corresponding liquid PEA concentrations of 6.3 x 10(1)-6.3 x 10(-6) (% v/v). We found that the threshold concentrations for PEA in LP (step 6.5) and PEA in MO (step 5.5) were significantly lower (P < 0.05) than for PEA in PG (step 4.0) and DPG (step 4.0) We then used GC to measure both the liquid and gas PEA concentrations for the dilution steps prepared with LP and PG. Although there were large threshold differences in the liquid concentrations of PEA in LP and PG, the headspace gas concentrations of PEA were the same. These results demonstrate the importance of determining the gas concentration of odorant stimuli when performing odor threshold measurements, in particular when comparing odor detection thresholds obtained using different solvents.     

Different thresholds for detection and discrimination of odors in the honey bee (Apis mellifera)

Naturally occurring odors used by animals for mate recognition, food identification and other purposes must be detected at concentrations that vary across several orders of magnitude. Olfactory systems must therefore have the capacity to represent odors over a large range of concentrations regardless of dramatic changes in the salience, or perceived intensity, of a stimulus. The stability of the representation of an odor relative to other odors across concentration has not been extensively evaluated. We tested the ability of honey bees to discriminate pure odorants across a range of concentrations at and above their detection threshold. Our study showed that pure odorant compounds became progressively easier for honey bees to discriminate with increasing concentration. Discrimination is, therefore, a function of odorant concentration. We hypothesize that the recruitment of sensory cell populations across a range of concentrations may be important for odor coding, perhaps by changing its perceptual qualities or by increasing its salience against background stimuli, and that this mechanism is a general property of olfactory systems.     

Visual wavelength discrimination by the loggerhead turtle, Caretta caretta

Marine turtles are visual animals, yet we know remarkably little about how they use this sensory capacity. In this study, our purpose was to determine whether loggerhead turtles could discriminate between objects on the basis of color. We used light-adapted hatchlings to determine the minimum intensity of blue (450 nm), green (500 nm), and yellow (580 nm) visual stimuli that evoked a positive phototaxis (the phototaxis "threshold" [pt]). Juvenile turtles were later trained to associate each color (presented at 1 log unit above that color's pt) with food, then to discriminate between two colors (the original rewarded stimulus plus one of the other colors, not rewarded) when both were presented at 1 log unit above their pt. In the crucial test, turtles were trained to choose between the rewarded and unrewarded color when the colors varied in intensity. All turtles learned that task, demonstrating color discrimination. An association between blue and food was acquired in fewer trials than between yellow and food, perhaps because some prey of juvenile loggerheads in oceanic surface waters (jellyfishes, polyps, and pelagic gastropods) are blue or violet in color.     

Olfactory sensitivity for aliphatic aldehydes in squirrel monkeys and pigtail macaques

Using a conditioning paradigm, the olfactory sensitivity of three squirrel monkeys and three pigtail macaques for a homologous series of aliphatic aldehydes ( n-butanal to n-nonanal) was assessed. With only few exceptions, the animals of both species significantly discriminated concentrations below 1 ppm from the odorless solvent, and with n-butanal and n-hexanal individual pigtail macaques even demonstrated thresholds below 1 ppb. The results showed (1). both primate species to have a well-developed olfactory sensitivity for aliphatic aldehydes, (2). pigtail macaques to generally perform better than squirrel monkeys in detecting members of this class of odorants, and (3). no significant correlation between perceptibility in terms of olfactory detection thresholds and carbon chain length of the aliphatic aldehydes in both species tested. These findings lend further support to the growing body of evidence suggesting that between-species comparisons of the number of functional olfactory receptor genes or of neuroanatomical features are poor predictors of olfactory performance. Further, our findings suggest that olfaction may play an important and hitherto underestimated role in the regulation of behavior in the species tested.     

Olfactory sensitivity for enantiomers and their racemic mixtures--a comparative study in CD-1 mice and spider monkeys

Using a conditioning paradigm, the olfactory sensitivity of six CD-1 mice for the enantiomers of carvone and of limonene as well as for their racemic mixtures was investigated. With all six stimuli, the animals significantly discriminated concentrations 相似文献   

Olfactory sensitivity and odor structure-activity relationships for aliphatic carboxylic acids in CD-1 mice

Using a conditioning paradigm, the olfactory sensitivity of CD-1 mice for a homologous series of aliphatic n-carboxylic acids (ethanoic acid to n-octanoic acid) and several of their isomeric forms was investigated. With all 14 odorants, the animals significantly discriminated concentrations as low as 0.03 ppm (parts per million) from the solvent, and with four odorants the best-scoring animals even detected concentrations as low as 3 ppt (parts per trillion). Analysis of odor structure-activity relationships showed that the correlation between olfactory detection thresholds of the mice for the unbranched carboxylic acids and carbon chain length can best be described as a U-shaped function with the lowest threshold values at n-butanoic acid. A significant positive correlation between olfactory detection thresholds and carbon chain length of the carboxylic acids with their branching next to the functional carboxyl group was found. In contrast, no such correlation was found for carboxylic acids with their branching at the distal end of the carbon chain relative to the functional carboxyl group. Finally, a significant correlation was found between olfactory detection thresholds and the position of the branching of the carboxylic acids. Across-species comparisons suggest that mice are more sensitive for short-chained (C(2) to C(4)) aliphatic n-carboxylic acids than other mammalian species, but not for longer-chained ones (C(5) to C(8)). Further comparisons suggest that odor structure-activity relationships are both substance class- and species-specific.     

Influences of feedback and ascending and descending trial presentations on perithreshold odor detection performance

The influences of feedback and ascending and descending trial sequences on the ability of 135 college-aged subjects to detect phenyl ethyl alcohol odorant concentrations ranging from 10(-9) to 10(-5.5) v/v were examined in a two-alternative forced-choice test paradigm. At the highest concentrations, ascending trial sequences produced better performance than descending trial sequences; the reverse was true at the lowest concentrations. There was a tendency for feedback to improve performance marginally at the lowest two odorant concentrations presented. In the region associated with a traditional detection threshold calculation (i.e. at the 75% performance point in a two-choice detection task), no influences of feedback or direction of trial sequence were apparent. These data indicate that the effects of explicit feedback and trial sequence direction depend upon the segment of the peri-threshold stimulus concentration continuum evaluated.     

Olfactory discrimination ability for aliphatic odorants as a function of oxygen moiety

We tested the ability of human subjects to distinguish between aliphatic odorants sharing the same number of carbon atoms but differing in their functional groups. 1-Alcohols, n-aldehydes, 2-ketones and n-carboxylic acids of four, six and eight carbon atoms, respectively, were employed. In a forced-choice triangular test procedure 20 subjects were repeatedly presented with 18 odor pairs and asked to identify the bottle containing the odd stimulus. We found (i) that as a group, the subjects performed significantly above chance level in all tasks and thus were clearly able to discriminate between all odor pairs presented; (ii) marked interindividual differences in discrimination performance, ranging from subjects who were able to significantly distinguish between all 18 odor pairs to subjects who failed to do so with 1/3 of the tasks; (iii) a lack of significant differences in performance between male and female, and between Japanese and German subjects; (iv) that odor pairs that involved 2-ketones and/or n-carboxylic acids were significantly easier to discriminate compared to odor pairs that involved 1-alcohols and/or n-aldehydes, and thus a clear dependence of discriminability on type of functional group; and (v) that aliphatic odorants with eight carbon atoms (irrespective of their oxygen moiety) were significantly more difficult to discriminate from each other compared to substances with four or six carbon atoms. The results suggest that functional groups may be an important determinant of the interaction between stimulus molecule and olfactory receptor in aliphatic substances, and thus may be a molecular property affecting odor quality in a substance class-specific manner.     

Asymmetric suppression of components in binary aldehyde mixtures: behavioral studies in the laboratory rat

The aim of the present study was to assess component interaction in the perception of the 2 aldehydes butanal and heptanal when presented in binary mixtures to rats. A further aim was to develop a behavioral paradigm for testing suppression of components in mixtures using rodent subjects. Thirsty rats were initially trained to discriminate between the 2 aldehydes butanal and heptanal in an olfactometer using a go/no-go discrimination task. This involved rats learning to place their noses in a sniff port where odors were presented and to lick a tube for water reward when one of the aldehydes was presented (S+) while withholding licking at the tube to the other, unrewarded, aldehyde (S-). A mixture condition was then introduced into the task, whereby a proportion of trials involved presentation of a combination of the 2 aldehydes as an additional unrewarded condition. Rats readily learned to withhold licking on trials when the mixture was presented. The concentration of the nonrewarded (S-) aldehyde in the mixture was then systematically decreased, whereas the concentration of the S+ component was held constant. This eventually caused the S+ component in the mixture to suppress detection of the S-, as shown by an increasing number of lick responses (false alarms) on trials when the mixture was presented. These suppressing effects occurred well above the detection threshold for the S- aldehyde presented alone. Results showed asymmetric suppression in the mixture condition such that butanal suppressed detection of heptanal at much lower concentrations than vice versa. A second experiment showed that when both butanal and heptanal were present in a binary mixture at the same concentration (10(-6) volume %), then rats responded to the mixture as if only butanal was present. These findings are discussed in terms of butanal having higher mobility and being able to compete more effectively than heptanal for occupation of shared receptor sites.     

Olfactory discrimination ability for aliphatic c6 alcohols as a function of presence, position, and configuration of a double bond

The ability of human subjects to distinguish between aliphatic C6 alcohols differing in presence, position, or configuration (i.e., cis-trans geometry) of a double bond was tested. In a forced-choice triangular test procedure, 20 subjects were repeatedly presented with all 21 binary combinations of the seven stimuli and asked to identify the bottle containing the odd stimulus. I found (a) that as a group, the subjects performed significantly above chance level in all tasks but two and thus were clearly able to discriminate between most of the odor pairs presented; (b) marked interindividual differences in discrimination performance, ranging from subjects who were able to significantly distinguish between all 21 odor pairs to subjects who failed to do so with 10 of the tasks; (c) that odor pairs involving two hexenols were significantly more difficult to discriminate than odor pairs that involved hexanol and one of the hexenols; (d) that odor pairs involving hexenols sharing the same geometry but differing in the position of the double bond by only one carbon atom were significantly more difficult to distinguish than odor pairs that involved hexenols differing by two carbon atoms; (e) that odor pairs involving 4-hexenols were significantly easier to discriminate than 3-hexenols, which, in turn, were significantly easier to distinguish than 2-hexenols; and (f) that odor pairs involving two cis-hexenols were significantly more difficult to discriminate than odor pairs that involved two trans-hexenols. These findings demonstrate that the presence as well as the position and configuration of a double bond affected discriminability in a systematic manner and suggest that these molecular structural features may be important determinants of the interaction between stimulus molecule and olfactory receptor and thus may affect odor quality of aliphatic alcohols.     

Complex mixture discrimination and the role of contaminants

Rats were trained in a 2-alternative odor choice task to discriminate between a 10-component odor mixture and the same mixture with one component removed and replaced with 1 of 3 concentrations of a different monomolecular odor (contaminant). All stimuli were presented within a training session, thus the rat essentially had to learn to discriminate the 10-component mixture from "not" the 10-component mixture. Rats performed most poorly discriminating the complete mixture from the mixture with one component removed and no contaminant added. As the concentration of the contaminant increased from 10 ppm to a concentration equal to the other components (100 ppm), discrimination improved linearly. In analyses of individual differences, rats that spent more time in the sampling port (sampling and making a decision) were more accurate than rats that spent less time. Together, these results emphasize the balance between perceptual stability and perceptual discrimination expressed by the olfactory system dealing with dynamic mixtures and the robust effects of contamination on those processes. In addition, they provide further support that modification of sampling/decision time is a strategy used by rats to deal with difficult discriminations of complex odors.     

Characterizing psychophysical measures of discrimination thresholds and the effects of concentration on discrimination learning in the moth Manduca sexta

What is the spatial and temporal nature of odor representations within primary olfactory networks at the threshold of an animal's ability to discriminate? Although this question is of central importance to olfactory neuroscience, it can only be answered in model systems where neural representations can be measured and discrimination thresholds between odors can be characterized. Here, we establish these thresholds for a panel of odors using a Pavlovian paradigm in the moth Manduca sexta. Moths were differentially conditioned to respond to one odor (CS+) but not another (CS-) using undiluted odorants to minimize salience-dependent learning effects. At 24 and 48 h postconditioning, moths were tested for the presence of a conditioned response (CR) with a blank, then the CS+ and CS- (pseudorandomly) across a 5-log step series of increasing concentration. Results identified discrimination thresholds and established that differential CRs to the CS+ and CS- increased with stimulus concentration. Next, 3 separate groups of moths were differentially conditioned at either one-log step below, at, or one log step above the identified discrimination threshold. At 24 and 48 h postconditioning, moths were tested sequentially with a blank, the concentration used for conditioning, and then undiluted odor. Conditioning at one log step below the discrimination threshold established a CR, indicating both stimulus detection and learning, but was insufficient to establish evidence of discrimination. Moths conditioned at the discrimination threshold were able to discriminate but only when stimulated with undiluted odors, indicating learning, but discrimination measures were hampered. When conditioned above the discrimination threshold, moths had no difficulty in discriminating. These results establish methods for psychophysical characterization of discrimination and indicate that differential conditioning at lowered concentrations biases threshold measures.     

Unitary Responses in Frog Olfactory Epithelium to Sterically Related Molecules at Low Concentrations

Responses of receptor cells in the frog's olfactory epithelium were recorded using platinum-black metal-filled microelectrodes. Spontaneous activity varied over a wide range from 0.07 to 1.8 spikes/s. Mean interspike intervals ranged from 13.7 to 0.5 s. Excitatory responses to six sterically related compounds at low concentrations were investigated. Stimuli were delivered in an aqueous medium. Thresholds for impulse initiation varied from greater than 1 mM down to the nanomolar concentration range. Thresholds of different olfactory receptors to the same stimulus could vary by several log units. Thresholds of the same receptor cell to different stimuli could be within the same order of magnitude, or could vary by as much as 5 log units. Based upon quantitative measures of stimulus-evoked excitatory responses it appeared that some receptors did not discriminate among sterically related molecules, whereas other receptors clearly discriminated between stimuli which evoke similar odor sensations.     

Human olfactory receptor families and their odorants

The human nose detects volatile chemical stimuli by at least three different receptor families: odorant receptors, trace amine-associated receptors, and vomeronasal type-1 receptors. As G protein-coupled receptors, all of the few functionally characterized olfactory receptors share major functional features: when expressed in heterologous cell systems, they 1) respond to odorants of certain chemical groups, e.g., amines, aliphatic carboxylic acids or aldehydes, floral or fruity odorants, including certain key-food odorants, and putative pheromones, and 2) transduce their signals to intracellular cAMP signaling. However, little is known yet about specific differences in the functional designation of the three olfactory receptor families. Recently, two heterologous cell systems expressing olfactory signaling molecules have been developed. Different screening strategies will shed light on the yet sparsely available odorant specificity profiles and structure-function relationships of olfactory receptors, as well as the structure-activity relationships of their odorants.     

Making sense of olfaction through predictions of the 3-D structure and function of olfactory receptors

We used the MembStruk first principles computational technique to predict the three-dimensional (3-D) structure of six mouse olfactory receptors (S6, S18, S19, S25, S46 and S50) for which experimental odorant recognition profiles are available for a set of 24 odorants (4-9 carbons aliphatic alcohols, acids, bromo-acids and diacids). We used the HierDock method to scan each predicted OR structure for potential odorant binding site(s) and to calculate binding energies of each odorant in these binding sites. The calculated binding affinity profiles are in good agreement with experimental activation profiles, validating the predicted 3-D structures and the predicted binding sites. For each of the six ORs, the binding site is located between trans-membrane domains (TMs) 3-6, with contributions from extracellular loops 2 and 3. In particular, we find six residue positions in TM3 and TM6 to be consistently involved in the binding modes of the odorants. Indeed, the differences in the experimental recognition profiles can be explained on the basis of these critical residues alone. These predictions are also consistent with mutation data on ligand binding for catecholamine receptors and sequence hypervariability studies for ORs. Based on this analysis, we defined amino acid patterns associated with the recognition of short aliphatic alcohols and mono-acids. Using these two sequence fingerprints to probe the alignment of 869 OR sequences from the mouse genome, we identified 34 OR sequences matching the fingerprint for aliphatic mono-acids and 36 corresponding to the recognition pattern for aliphatic alcohols. We suggest that these two sets of ORs might function as basic arrays for uniquely recognizing aliphatic alcohols and acids. We screened a library of 89 additional molecules against the six ORs and found that this set of ORs is likely to respond to aldehydes and esters with longer carbon chains than their currently known agonists. We also find that compounds associated with the flavor in foods are often among the best calculated binding affinities. This suggests that physiologic ligands for these ORs may be found among aldehydes and esters associated with flavor.     

Response patterns of single neurons in the tortoise olfactory epithelium and olfactory bulb

The responses to odor stimulation of 40 single units in the olfactory mucosa and of 18 units in the olfactory bulb of the tortoise (Gopherus polyphemus) were recorded with indium-filled, Pt-black-tipped microelectrodes. The test battery consisted of 27 odorants which were proved effective by recording from small bundles of olfactory nerve. Two concentrations of each odorant were employed. These values were adjusted for response magnitudes equal to those for amyl acetate at –2.5 and –3.5 log concentration in olfactory twig recording. Varying concentrations were generated by an injection-type olfactometer. The mucosal responses were exclusively facilitory with a peak frequency of 16 impulses/sec. 19 mucosal units responded to at least one odorant and each unit was sensitive to a limited number of odorants (1–15). The sensitivity pattern of each unit was highly individual, with no clear-cut types, either chemical or qualitative, emerging. Of the 18 olfactory bulb units sampled, all responded to at least one odorant. The maximum frequency observed during a response was 39 impulses/sec. The bulbar neurons can be classified into two types. There are neurons that respond exclusively with facilitation and others that respond with facilitation to some odorants and with inhibition to others. Qualitatively or chemically similar odorants did not generate similar patterns across bulbar units.     

Relation of spatial and temporal frequencies to vernier acuity

Vernier thresholds were measured with a pair of vertical sinusoidal gratings of one and a half cycles as targets. The amplitude was weighted by a one-dimensional Gaussian and contrast was set one log unit above contrast threshold. The vernier thresholds were estimated with the method of constant stimuli. Temporal frequency effects were introduced by movement of the vernier targets. It was found that vernier thresholds expressed in phase angle were unchanged in the effective range of spatial frequencies provided that the temporal frequency and the visibility were unchanged, and that thresholds deteriorated by increasing the temporal frequency. It is suggested that the detection of relative phase may be involved in the discrimination of vernier offsets and that it may be mediated by a sustained unit. Three possible types of mechanisms, edge-localization processes, orientation-selective units and phase-sensitive units, were considered in relation to vernier acuity.