Real-time monitoring of the metabolic capacity of ex vivo rat olfactory mucosa by proton transfer reaction mass spectrometry (PTR-MS).

Olfactory mucosa (OM) can metabolise odorant volatile organic compounds through various enzymatic mechanisms to produce odorous or non-odorous metabolites. Preliminary ex vivo studies using headspace-gas chromatography (HS-GC) revealed the formation of metabolites when odorant molecules were injected in the headspace above a fresh explant of rat olfactory mucosa. However, this method did not allow accessing the data during the first 5 min of contact between the odorant and the mucosa; thus limiting the olfactory biological significance. Using a direct-injection mass spectrometry technique with a proton transfer reaction instrument (PTR-MS), we have been able, for the first time, to investigate the first moments of the enzymatic process of the metabolic capacity of ex vivo rat olfactory mucosa in real time. Using ethyl acetate as a model volatile odorous substrate, we demonstrated here for the first time that this odorant could be metabolised by an ex vivo olfactory mucosa within seconds, producing ethanol as metabolite.

Investigating the human chemical communication of positive emotions using a virtual reality-based mood induction.

Humans can communicate their emotions to others via volatile emissions from their bodies. Although there is now solid evidence for human chemical communication of fear, stress and anxiety, investigations of positive emotions remain scarce. In a recent study, we found that women's heart rate and performance in creativity tasks were modulated by body odors of men sampled while they were in a positive vs. neutral mood. However, inducing positive emotions in laboratory settings remains challenging. Therefore, an important step to further investigate the human chemical communication of positive emotions is to develop new methods to induce positive moods. Here, we present a new mood induction procedure (MIP) based on virtual reality (VR), that we assumed to be more powerful than videos (used in our previous study) to induce positive emotions. We hypothesized that, consequently, given the more intense emotions created, this VR-based MIP would induce larger differences between the receivers' responses to the positive body odor versus a neutral control body odor, than the Video-based MIP. The results confirmed the higher efficacy of VR to induce positive emotions compared with videos. More specifically, VR had more repeatable effects between individuals. Although positive body odors had similar effects to those found in the previous video study, especially faster problem solving, these effects did not reach statistical significance. These outcomes are discussed as a function of the specificities of VR and of other methodological parameters, that may have prevented the observation of such subtle effects and that should be understood more in-depth for future studies on human chemical communication.

Nasal Odorant Competitive Metabolism Is Involved in the Human Olfactory Process.

Within the peripheral olfactory process, odorant metabolizing enzymes are involved in the active biotransformation of odorants, thus influencing the intensity and quality of the signal, but little evidence exists in humans. Here, we characterized the fast nasal metabolism of the food aroma pentane-2,3-dione in vivo and identified two resulting metabolites in the nasal-exhaled air, supporting the metabolizing role of the dicarbonyl/l-xylulose reductase. We showed in vitro, using the recombinant enzyme, that pentane-2,3-dione metabolism was inhibited by a second odorant (e.g., butanoic acid) according to an odorant-odorant competitive metabolic mechanism. Hypothesizing that such mechanism exists in vivo, pentane-2,3-dione, presented with a competitive odorant, both at subthreshold concentrations, was actually significantly perceived, suggesting an increase in its nasal availability. Our results, suggesting that odorant metabolizing enzymes can balance the relative detection of odorants in a mixture, in turn influencing the intensity of the signal, should be considered to better manage flavor perception in food.

Ex vivo real-time monitoring of volatile metabolites resulting from nasal odorant metabolism.

Odorant-metabolizing enzymes are critically involved in the clearance of odorant molecules from the environment of the nasal neuro-olfactory tissue to maintain the sensitivity of olfactory detection. Odorant metabolism may also generate metabolites in situ, the characterization and function of which in olfaction remain largely unknown. Here, we engineered and validated an ex vivo method to measure odorant metabolism in real-time. Glassware containing an explant of rat olfactory mucosa was continuously flushed with an odorant flow and was coupled to a proton transfer reaction-mass spectrometer for volatile compound analysis. Focusing on carboxylic esters and diketone odorants, we recorded the metabolic uptake of odorants by the mucosa, concomitantly with the release of volatile odorant metabolites in the headspace. These results significantly change the picture of real-time in situ odorant metabolism and represent a new step forward in the investigation of the function of odorant metabolites in the peripheral olfactory process. Our method allows the systematic identification of odorant metabolites using a validated animal model and permits the screening of olfactory endogenously produced chemosensory molecules.

Nasal mucus glutathione transferase activity and impact on olfactory perception and neonatal behavior.

In olfaction, to preserve the sensitivity of the response, the bioavailability of odor molecules is under the control of odorant-metabolizing enzymes (OMEs) expressed in the olfactory neuroepithelium. Although this enzymatic regulation has been shown to be involved in olfactory receptor activation and perceptual responses, it remains widely underestimated in vertebrates. In particular, the possible activity of OMEs in the nasal mucus, i.e. the aqueous layer that lined the nasal epithelium and forms the interface for airborne odorants to reach the olfactory sensory neurons, is poorly known. Here, we used the well-described model of the mammary pheromone (MP) and behavioral response in rabbit neonates to challenge the function of nasal mucus metabolism in an unprecedented way. First, we showed, in the olfactory epithelium, a rapid glutathione transferase activity toward the MP by ex vivo real-time mass spectrometry (PTR-MS) which supported an activity in the closest vicinity of both the odorants and olfactory receptors. Indeed and second, both the presence and activity of glutathione transferases were evidenced in the nasal mucus of neonates using proteomic and HPLC analysis respectively. Finally, we strikingly demonstrated that the deregulation of the MP metabolism by in vivo mucus washing modulates the newborn rabbit behavioral responsiveness to the MP. This is a step forward in the demonstration of the critical function of OMEs especially in the mucus, which is at the nasal front line of interaction with odorants and potentially subjected to physiopathological changes.

Publisher Correction: Nasal mucus glutathione transferase activity and impact on olfactory perception and neonatal behavior.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Publisher Correction: Ex vivo real-time monitoring of volatile metabolites resulting from nasal odorant metabolism.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Odorant-odorant metabolic interaction, a novel actor in olfactory perception and behavioral responsiveness.

In the nasal olfactory epithelium, olfactory metabolic enzymes ensure odorant clearance from the olfactory receptor environment. This biotransformation of odorants into deactivated polar metabolites is critical to maintaining peripheral sensitivity and perception. Olfactory stimuli consist of complex mixtures of odorants, so binding interactions likely occur at the enzyme level and may impact odor processing. Here, we used the well-described model of mammary pheromone-induced sucking-related behavior in rabbit neonates. It allowed to demonstrate how the presence of different aldehydic odorants efficiently affects the olfactory metabolism of this pheromone (an aldehyde too: 2-methylbut-2-enal). Indeed, according to in vitro and ex vivo measures, this metabolic interaction enhances the pheromone availability in the epithelium. Furthermore, in vivo presentation of the mammary pheromone at subthreshold concentrations efficiently triggers behavioral responsiveness in neonates when the pheromone is in mixture with a metabolic challenger odorant. These findings reveal that the periphery of the olfactory system is the place of metabolic interaction between odorants that may lead, in the context of odor mixture processing, to pertinent signal detection and corresponding behavioral effect.