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.     

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.     

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 ability of human subjects for ten pairs of enantiomers.

We tested the ability of human subjects to distinguish between enantiomers, i.e. odorants which are identical except for chirality. In a forced-choice triangular test procedure 20 subjects were repeatedly presented with 10 enantiomeric odor pairs and asked to identify the bottle containing the odd stimulus. We found (i) that as a group, the subjects were only able to significantly discriminate the optical isomers of alpha-pinene, carvone and limonene, whereas they failed to distinguish between the (+)- and (-)-forms of menthol, fenchone, rose oxide, camphor, alpha-terpineol, beta-citronellol and 2-butanol; (ii) marked individual differences in discrimination performance, ranging from subjects who were able to significantly discriminate between 6 of the 10 odor pairs to subjects who failed to do so with 9 of the 10 tasks; (iii) that with none of the 10 odor pairs were the antipodes reported to differ significantly in subjective intensity when presented at equal concentrations; and (iv) that error rates were quite stable and did not differ significantly between sessions, and thus, we observed a lack of learning or training effects. Additional tests of the degree of trigeminality and threshold measurements of the optical isomers of alpha-pinene, carvone and limonene suggest that the discriminability of these three enantiomeric odor pairs is indeed due to differences in odor quality. These findings support the assumption that enantioselective molecular odor receptors may only exist for some but not all volatile enantiomers and thus that chiral recognition of odorants may not be a general phenomenon but is restricted to some substances.     

Olfactory discrimination ability and odor structure-activity relationships in honeybees.

Using the training procedure introduced by von Frisch in 1919, we tested the ability of free-flying honeybees to discriminate a conditioning odor from an array of 44 simultaneously presented substances. The stimuli included homologous series of aliphatic alcohols, aldehydes and ketones, isomeric forms of some of these substances, as well as several terpenes and odor mixtures, and thus comprised stimuli of varying degrees of structural similarity to any conditioning odor. We found (i) that the honeybees significantly distinguished between 97.0% of the 1848 odor pairs tested, thus showing an excellent discrimination performance when tested in a free-flying situation with an array of structurally related substances; (ii) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with all aliphatic substance classes tested; (iii) that both the position and type of a functional group also affected discriminability of odorants in a substance class-specific manner; and (iv) striking similarities in odor structure-activity relationships between honeybees and human and nonhuman primates tested previously on a subset of substances employed here. Our findings demonstrate that the similarities found in the structural organization of the olfactory systems of insects and vertebrates are paralleled by striking similarities in relative discrimination abilities. This strongly suggests that similar mechanisms of odor coding and discrimination may underlie olfaction in vertebrates and insects.     

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.     

Olfactory perception in honeybees: Concatenated and mixed odorant stimuli,concentration, and exposure effects

Summary Here we present results obtained from 7 different series of experiments, all employing odor conditioning of proboscis extension in worker honeybees and each designed to address a particular question involving olfactory perception. The questions relate to: temporal complexity of odor cues; effects of concentration, suppression, and/or potentiation in mixture perception; acquisition and extinction rates, as well as levels of generalization associated with aliphatic compounds that have the same functional groups or same alkyl radical length; and the effects of continuous exposure to odorants in the first several days of adult life on various learning and discrimination tasks involving olfactory perception. From the data obtained in these experiments we were able to conclude the following: First, worker honeybees have a limited ability to perceive complex temporal odor-quality patterns in olfactory stimuli — they learn to associate the quality of only the last part of the stimulus with a sucrose reward. Second, we confirm that citral is qualitatively different in several perceptual contexts involving odor learning and conditioning and our results help elucidate the nature of these differences as they relate to learning, discrimination, mixture perception, and continuous exposure to particular odorants. Third, we appear to have uncovered some important perceptual differences between functional groups attached to the first as opposed to the second carbon atom of alkyl radicals. Finally, we failed to uncover any significant effects relating to continuous exposure to odorants during the first several days of a worker's adult life, despite evidence that considerable sensory development takes place during this period. Thus ontogenetic changes to the peripheral system due to environmental effects appear to leave basic perceptual systems unaltered.     

Olfactory Discrimination Ability for Aliphatic Esters in Squirrel Monkeys and Humans

Using a behavioral paradigm designed to simulate olfactory-guidedforaging, the ability of five squirrel monkeys to distinguishiso-amyl acetate from n-and iso-forms of other acetic esters(ethyl acetate to decyl acetate) and from other esters carryingthe iso-amyl group (iso-amyl propionate to iso-amyl capronate)was investigated. We found (i) that all five animals were clearlyable to discriminate between all odor pairs tested; (ii) a significantnegative correlation between discrimination performance andstructural similarity of odorants in terms of differences incarbon chain length of both the aliphatic alcohol group andthe aliphatic acid group of the esters; and (iii) that iso-and n-amyl acetate were perceived as qualitatively similar despitedifferent steric conformation. Using a triple-forced choiceprocedure, 20 human subjects were tested on the same tasks inparallel and showed a very similar pattern of discriminationperformance compared with the squirrel monkeys. Thus, the resultsof this study provide evidence of well-developed olfactory discriminationability in squirrel monkeys for aliphatic esters and supportthe assumption that human and non-human primates may share commonprinciples of odor quality perception. Chem. Senses 22: 457–465,1997.     

Olfactory-based discrimination learning in the moth, Manduca sexta

Moths possess highly tuned olfactory capabilities, which can detect very low concentrations of pheromonal odorants. Much is known about the structure and function of the moth olfactory system with respect to detection of pheromones. However, we lack an understanding of the broader olfactory system, in particular, to what degree are moths capable of detecting and discriminating odorants that are not components of pheromone blends. Here we describe a methodology used to investigate the discriminability of nonpheromonal odors in moths. In a series of experiments we show that the moth Manduca sexta can (1) discriminate a number of different odors but (2) that methyl jasmonate, neither readily conditions to a food reward nor is it readily discriminated from another odor. The lack of a response to methyl jasmonate may be related to its role in host plant defense. This work provides a basis for future mapping of physiological and pharmacological studies of nonpheromonal coding in insects onto learned behavioral responses to those odorants.     

Dissociable codes of odor quality and odorant structure in human piriform cortex

The relationship between odorant structure and odor quality has been a focus of olfactory research for 100 years, although no systematic correlations are yet apparent. Animal studies suggest that topographical representations of odorant structure in olfactory bulb form the perceptual basis of odor quality. Whether central olfactory regions are similarly organized is unclear. Using an olfactory version of fMRI cross-adaptation, we measured neural responses in primary olfactory (piriform) cortex as subjects smelled pairs of odorants systematically differing in quality and molecular functional group (as one critical attribute of odorant structure). Our results indicate a double dissociation in piriform cortex, whereby posterior regions encode quality (but not structure) and anterior regions encode structure (but not quality). The presence of structure-based codes suggests fidelity of sensory information arising from olfactory bulb. In turn, quality-based codes are independent of any simple structural configuration, implying that synthetic mechanisms may underlie our experience of smell.     

Olfactory discrimination ability of human subjects for enantiomers with an isopropenyl group at the chiral center

The ability of 20 human subjects to distinguish between nine enantiomeric odor pairs sharing an isopropenyl group at the chiral center was tested in a forced-choice triangular test procedure. I found (i). that as a group, the subjects were only able to significantly discriminate the optical isomers of limonene, carvone, dihydrocarvone, dihydrocarveol and dihydrocarvyl acetate, whereas they failed to distinguish between the (+)- and (-)-forms of perillaalcohol, perillaaldehyde, isopulegol and limonene oxide; (ii). marked interindividual differences in discrimination performance, ranging from subjects who were able to significantly discriminate between eight of the nine odor pairs to subjects who failed to do so with six of the nine tasks; and (iii). that with none of the nine odor pairs the antipodes were reported to differ significantly in subjective intensity when presented at equal concentrations. Additional tests of the chemesthetic potency and threshold measurements of the optical isomers of dihydrocarvone, dihydrocarveol, and dihydrocarvyl acetate suggest that the discriminability of these three enantiomeric odor pairs is indeed due to differences in odor quality. Analysis of structure-activity relationships suggest that the combined presence of (i). an isopropenyl group at the chiral center; (ii). a methyl group at the para-position; and/or (iii). an oxygen-containing group at the meta-position allows for the discrimination of enantiomeric odor pairs.     

Concentration-dependent changes of perceived odor quality

In order to assess the dependence of perceived odor qualityon odorant concentration, we studied 21 subjects. For eightsubjects all possible pairs from a pool of six odorants at threedecimal dilutions were presented, and subjects were requestedto state whether members of the pair were qualitatively ‘similar’or ‘different’ It was found that while pairs withthe same odorant at identical concentrations were judged ‘similar’in >90% of the cases by all subjects, scores went down to10% ‘similar’ judgements in some cases when thesame odorant was presented at a 100-fold concentration difference.Large time-invariable differences were found among subjectsand among odorants. For the additional 13 subjects, all possiblepairs from a pool of four odorants at three decimal dilutionswere presented. Subjects were instructed to state whether membersof the pair were qualitatively ‘same’ or ‘different’,and were also requested to rank the degree of difference ona visual analogue scale. Results for this group were, in general,similar to the results of the former group of subjects and goodagreement between the two tasks was found. The results suggestthat variations in olfactory stimulus magnitude may be perceivedas quality differences, as previously shown for vision and audition.     

Odorant moiety and odor mixture perception in free-flying honey bees (Apis mellifera)

Two experiments are described that employ a Y-tube odor-trainingparadigm to address questions relating to olfactory perceptionin free-flying worker honey bees. The first is designed to evaluatehow easily bees can be conditioned to discriminate between twoodors and how willing they are to generalize between closelyrelated odors. In particular, we demonstrate that individualworker bees have no trouble learning to discriminate betweenalkyl ketones or alcohols that differ by only one carbon atom(e.g. heptanone versus octanone) or between a ketone and alcoholfunctional group attached to the same alkyl radical; but theygeneralize between compounds with the same functional groupmuch more readily than those with the same alkyl radical. Thesecond experiment is designed to explore the relationship betweenthe perception of a mixture of odorants and the perception ofthe individual odorants themselves. Our results suggest thatthere appears to be a stronger relationship between a two-odorantmixture and its constituents than would be suggested by themixture being an odor intermediate between the two constituentodorants. We also include a comprehensive discussion on theproblem of extracting quality and concentration informationfrom an odor stimulus and we explore ideas relating to the perceptionof the constituent odorant components of complex odors.     

Concentration and Membrane Fluidity Dependence of Odor Discrimination in the Turtle Olfactory System

In the present study, we examined the concentration dependenceof odor discrimination in turtle olfactory bulbar responsesusing the cross-adaptation technique. In the odorant pairs withdiverse molecular structures, the degree of discrimination wasunchanged or only slightly decreased with an increase in odorantconcentrations, suggesting that odorants are well discriminatedeven at high concentrations. In the odorant pairs with closelyrelated molecular structures, the degree of discrimination wasdecreased with an increase in odorant concentrations. An increasein the temperature of turtle olfactory epithelium also decreasedthe ability to discriminate these odorants. There was a goodcorrelation between changes in the odor discriminating abilityinduced by an increase in odor concentrations and those inducedby a temperature increase. The liposomes were made of lipidsextracted from the turtle olfactory epithelia and changes oftheir membrane fluidity induced by adsorption of odorants weremonitored with DPH. There was a good correlation between a decreasein odor discriminating ability and the membrane fluidity changesinduced by odorants. We suggest that decreases in odor discriminatingability induced either by an increase in odor concentrationor by a temperature increase are ultimately caused by changesin the membrane fluidity. Chem. Senses 22: 553–563, 1997.     

Intranasal localizability of odorants: influence of stimulus volume

When an odorant is presented to one side of the nose and air to the other, the ability to localize which side received the odorant depends upon trigeminal nerve stimulation. It has been shown that performance on this lateralization task increases as stimulus concentration increases. In this study, we determined the influences of stimulus volume and sex on the ability to localize each of 8 odorants presented at neat concentrations: anethole, geraniol, limonene, linalool, menthol, methyl salicylate, phenyl ethanol, and vanillin. At a low stimulus volume (11 mL), only menthol was localized at an above-chance level. At a high stimulus volume (21 mL), above-chance localization occurred for all odorants except vanillin. Women were significantly better than men in localizing menthol. Stimuli rated as most intense were those that were most readily localized. The detection performance measures, as well as rated intensity values, significantly correlated with earlier findings of the trigeminal detectability of odorants presented to anosmic and normosmic subjects. This study suggests that differences in stimulus volume may explain some discrepant findings within the trigeminal chemosensory literature and supports the concept that vanillin may be a "relatively pure" olfactory stimulus.     

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.     

'Sniffin' Sticks': Olfactory Performance Assessed by the Combined Testing of Odour Identification, Odor Discrimination and Olfactory Threshold

‘Sniffin’ Sticks' is a new test of nasal chemosensoryperformance based on pen-like odor dispensing devices. It comprisesthree tests of olfactory function, namely tests for odor threshold(n-butanol, testing by means of a single staircase), odor discrimination(15 pairs of odorants, triple forced choice) and odor identification(16 common odorants, multiple forced choice from four verbalitems per test odorant). After extensive preliminary investigationsthe tests were applied to a group of 104 healthy volunteers(52 female, 52 male, mean age 49.5 years, range 18–84years) in order to establish test-retest reliability and tocompare them with an established measure of olfactory performance(the Connecticut Chemosensory Clinical Research Center Test,CCCRC). Performance decreased with increasing age of the subjects(     

Developmental regulation of olfactory circuit formation in mice

In mammals, odorants induce various behavioral responses that are critical to the survival of the individual and species. Binding signals of odorants to odorant receptors (ORs) expressed in the olfactory epithelia are converted to an odor map, a pattern of activated glomeruli, in the olfactory bulb (OB). This topographic map is used to identify odorants for memory-based learned decisions. In the embryo, a coarse olfactory map is generated in the OB by a combination of dorsal-ventral and anterior-posterior targeting of olfactory sensory neurons (OSNs), using specific sets of axon-guidance molecules. During the process of OSN projection, odor signals are sorted into distinct odor qualities in separate functional domains in the OB. Odor information is then conveyed by the projection neurons, mitral/tufted cells, to various regions in the olfactory cortex, particularly to the amygdala for innate olfactory decisions. Although the basic architecture of hard-wired circuits is generated by a genetic program, innate olfactory responses are modified by neonatal odor experience in an activity-dependent manner. Stimulus-driven OR activity promotes post-synaptic events and dendrite selection in the responding glomeruli making them larger. As a result, enhanced odor inputs in neonates establish imprinted olfactory memory that induces attractive responses in adults, even when the odor quality is innately aversive. In this paper, I will provide an overview of the recent progress made in the olfactory circuit formation in mice.     

Learning to smell the roses: experience-dependent neural plasticity in human piriform and orbitofrontal cortices

It is widely presumed that odor quality is a direct outcome of odorant structure, but human studies indicate that molecular knowledge of an odorant is not always sufficient to predict odor quality. Indeed, the same olfactory input may generate different odor percepts depending on prior learning and experience. Combining functional magnetic resonance imaging with an olfactory paradigm of perceptual learning, we examined how sensory experience modifies odor perception and odor quality coding in the human brain. Prolonged exposure to a target odorant enhanced perceptual differentiation for odorants related in odor quality or functional group, an effect that was paralleled by learning-induced response increases in piriform cortex and orbitofrontal cortex (OFC). Critically, the magnitude of OFC activation predicted subsequent improvement in behavioral differentiation. Our findings suggest that neural representations of odor quality can be rapidly updated through mere perceptual experience, a mechanism that may underlie the development of odor perception.     

Molecular, anatomical, and functional organization of the Drosophila olfactory system

BACKGROUND: Olfactory receptor neurons (ORNs) convey chemical information into the brain, producing internal representations of odors detected in the periphery. A comprehensive understanding of the molecular and neural mechanisms of odor detection and processing requires complete maps of odorant receptor (Or) expression and ORN connectivity, preferably at single-cell resolution. RESULTS: We have constructed near-complete maps of Or expression and ORN targeting in the Drosophila olfactory system. These maps confirm the general validity of the "one neuron--one receptor" and "one glomerulus--one receptor" principles and reveal several additional features of olfactory organization. ORNs in distinct sensilla types project to distinct regions of the antennal lobe, but neighbor relations are not preserved. ORNs grouped in the same sensilla do not express similar receptors, but similar receptors tend to map to closely appositioned glomeruli in the antennal lobe. This organization may serve to ensure that odor representations are dispersed in the periphery but clustered centrally. Integrated with electrophysiological data, these maps also predict glomerular representations of specific odorants. Representations of aliphatic and aromatic compounds are spatially segregated, with those of aliphatic compounds arranged topographically according to carbon chain length. CONCLUSIONS: These Or expression and ORN connectivity maps provide further insight into the molecular, anatomical, and functional organization of the Drosophila olfactory system. Our maps also provide an essential resource for investigating how internal odor representations are generated and how they are further processed and transmitted to higher brain centers.