Selective and reversible blockage of a fatty acid odour response in the olfactory bulb of the frog (Rana temporaria).

The lectin concanavalin A (ConA) when applied to the olfactory mucosa (OM) of frog and rat, is reported to partially inhibit electro-olfactogram (EOG) responses to fatty acid odours. Control odours like isoamyl acetate were not affected. We have now studied in the frog whether this treatment affects the corresponding olfactory bulb (OB) response. The OB surface was impregnated with a voltage-sensitive dye (RH 414). Spatial and temporal patterns of odour response were measured by changes in dye fluorescence that occur when OB neurons fire. The apparatus, consisted of an epi-fluorescent microscope coupled to a 64 x 64 pixel CCD photodetection camera. This allowed imaging over an 0.9 mm2 area of the OB glomerular layer to high resolution. When the frog OM was bathed with 5 mg ml(-1) ConA in Ringer's solution, the n-butyric acid odour response in the OB largely disappeared while the isoamyl acetate response did not change. When this experiment was repeated in the presence of 20 mM methyl alpha-D mannopyranoside (a ConA inhibitor), ConA failed to inhibit the n-butyric acid response. Moreover the ConA effect was partially reversible. A Ringer's wash of the OM after ConA treatment, partially restored the OB response to n-butyric acid. Thus the olfactory bulb results seem compatible with the EOG results and reinforce the notion that ConA selectively prevents n-butyric acid sensitive olfactory receptor neurons from firing. Chemical modification of the OM and their effect on OB response patterns may provide a useful approach to investigate olfactory quality coding.

Selective and reversible reduction of odor sensitivity in the rat by concanavalin A.

Rats were trained using an olfactometer to detect low concentrations of ethyl acetate (EA) or dimethyl disulphide vapor (DMDS). Concanavalin A (ConA) applied to the olfactory mucosa had no effect on ability of rats to detect low concentrations of EA but produced a marked deficit in detection in DMDS. DMDS detection performance returned to control levels 3 days after the ConA treatment. These results provide the first behavioral evidence for odor-specific inhibition by chemical modification of the olfactory epithelium and support the notion that ConA selectively inactivates one or more types (subclasses) of olfactory receptor proteins.

Concanavalin A reveals olfactory receptors which discriminate between alkane odorants on the basis of size.

For certain odorants, the amplitude of the rat electro-olfactogram is reduced if the olfactory epithelium is treated with the lectin concanavalin A. When normal and cycloalkanes of one to ten carbon atoms are used as odorants at equimolar concentration, the maximum reduction in amplitude is found to correlate with the size of the stimulus molecule. This observation is consistent with the notion that concanavalin A disables an olfactory receptor molecule which normally responds to the alkyl moiety of odorants in a particular size range. That moiety may thus represent a 'primary' quality-determining component in odour discrimination.

Sensory evidence for olfactory receptors with opposite chiral selectivity.

Human odor responses to alpha-ionone enantiomers were measured by psycho-physical methods. Surprisingly, subjects' relative sensitivities for these enantiomers were found to diverge widely, some subjects being much more sensitive to (+) than to (-) and vice versa. In 63 subjects the (+)/(-) threshold concentration ratio varied over 4 orders of magnitude. Nine subjects with extreme (+)/(-) ratios were further tested several times over a 7-month period and retained stable ratios. These same nine subjects, when tested for threshold sensitivity to carvone enantiomers, showed (+)/(-) ratios that varied only in a narrow range and did not correlate with their ionone ratios. It is suggested that odor discrimination of alpha-ionone enantiomers involves at least two receptor types of opposite chiral selectivity and that their distribution varies independently in the human population.

The effect of concanavalin A on the rat electro-olfactogram. Differential inhibition of odorant response.

When the rat olfactory mucosa is treated with concanavalin A, it subsequently shows diminished sensitivity towards 60% of the 112 odorants tested (as judged by the amplitude of the electro-olfactogram response). Odorants containing four to six carbon atoms tend to show the largest (absolute) diminutions, suggesting a receptor for this kind of odorant, although the structural specificity is weak. The receptor seems to be of particular importance in the detection of thiols, carboxylic acids and hydrocarbons of the above size, since these compounds loose the highest proportion of their original signal. The concanavalin A appears to be binding to the glycan of one or more cell-surface proteins. The binding may be at, or close to, at least one odorant receptor.

The effect of concanavalin A on the rat electro-olfactogram at various odorant concentrations.

We have studied the effect of concanavalin A (Con A) on the rat electro-olfactogram response to several odorants. Each odorant was applied over a range of concentrations. For hydrophobic odorants whose response was affected by Con A, the diminution in response was maximal at odorant concentrations of about 1 microM in the olfactory mucus. The (odour) concentration-dependence of the change is compatible with the idea that Con A inactivates one or more types of olfactory receptor that normally bind odorants with dissociation constants of the order of 100 nM. With hydrophilic odorants we had to apply concentrations very much higher than this to elicit any response from the system. At these high concentrations we could observe Con A-induced diminutions in response.

Odor similarity between stress-inducing odorants in wistar rats.

Odor similarity to the odor of 4-mercapto-4-methyl-2-pentanone (I) was measured on eight rats by generalization of a conditioned avoidance response tocis- andtrans-8-mercapto-p-menthan-3-one (II and III), 3-mercapto-3-methyl-2-pentanone (V), andt-amyl mercaptan (VI). Previously, these odorants had been found to induce stress in rats in an open-field situation. In the present experiment, rats generalized the avoidance response learned with I, for V, VI, and to a lesser extenttrans-isomer III, implying odor similarities;cis isomer II was discriminated. Odor similarity between mercapto ketone I and mercaptan VI is surprising since VI lacks the keto group.

Structure-activity relationship of stress-inducing odorants in the rat.

The stress for 12 sulfur-containing synthetized volatiles was evaluated in male Wistar rats and compared to that for fox-dropping extract concentrate. Stress behavior was analyzed by quantifying various stress responses in a standard open field and measuring the increase in plasma corticosterone concentration. Nine compounds induced stress-a dihydrothiazole, two cyclic polysulfides, five mercaptoketones, and a mercaptan. For the mercaptoketones, the following structure-activity relationships were observed. Size can vary considerably; the mercapto group can be either alpha or beta and either secondary or tertiary. The keto group is not essential, since a structurally related mercaptan remains active. The mercapto group is essential for activity in mercaptoketones, since conversion to a methyl sulfide resulted in a neutral response. This type of odorant could function as an allomone and may have potential in rat control as an area repellent.