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Detection and discrimination of odorants by the olfactory system plays a pivotal role in animal survival. Olfactory-based behaviors must be adapted to an ever-changing environment. Part of these adaptations includes changes of odorant detection by olfactory sensory neurons localized in the olfactory epithelium. It is now well established that internal signals such as hormones, neurotransmitters, or paracrine signals directly affect the electric activity of olfactory neurons. Furthermore, recent data have shown that activity-dependent survival of olfactory neurons is important in the olfactory epithelium. Finally, as olfactory neurons are directly exposed to environmental toxicants and pathogens, the olfactory epithelium also interacts closely with the immune system leading to neuroimmune modulations. Here, we review how detection of odorants can be modulated in the vertebrate olfactory epithelium. We choose to focus on three cellular types of the olfactory epithelium (the olfactory sensory neuron, the sustentacular and microvillar cells) to present the diversity of modulation of the detection of odorant in the olfactory epithelium. We also present some of the growing literature on the importance of immune cells in the functioning of the olfactory epithelium, although their impact on odorant detection is only just beginning to be unravelled.

Neuropeptide Y (NPY) plays an important role in regulating appetite and hunger in vertebrates. In the hypothalamus, NPY stimulates food intake under the control of the nutritional status. Previous studies have shown the presence of NPY and receptors in rodent olfactory system, and suggested a neuroproliferative role. Interestingly, NPY was also shown to directly modulate olfactory responses evoked by a food-related odorant in hungry axolotls. We have recently demonstrated that another nutritional cue, insulin, modulates the odorant responses of the rat olfactory mucosa (OM). Therefore, the aim of the present study was to investigate the potential effect of NPY on rat OM responses to odorants, in relation to the animal's nutritional state. We measured the potential NPY modulation of OM responses to odorant, using electro-olfactogram (EOG) recordings, in fed and fasted adult rats. NPY application significantly and transiently increased EOG amplitudes in fasted but not in fed rats. The effects of specific NPY-receptor agonists were similarly quantified, showing that NPY operated mainly through Y1 receptors. These receptors appeared as heterogeneously expressed by olfactory neurons in the OM, and western blot analysis showed that they were overexpressed in fasted rats. These data provide the first evidence that NPY modulates the initial events of odorant detection in the rat OM. Because this modulation depends on the nutritional status of the animal, and is ascribed to NPY, the most potent orexigenic peptide in the central nervous system, it evidences a strong supplementary physiological link between olfaction and nutritional processes.

All olfactory epithelium cells, including rapidly self-renewing olfactory sensory neurons (OSN), are continuously subjected to external airborne aggressions. We hypothesized that the apical part of rat olfactory epithelia (AOE) could be the site of a local translation to be able to respond rapidly to external stimuli. We purified significant amounts of mRNAs from AOE. Sequencing of the cDNA library identified 348 mRNA species. Of these, the 220 AOE transcripts encoding proteins with known biological functions were classified in functional groups. The main functional class (40%) coded for defense, detoxification, anti-oxidant stress and innate immunity. Other classes comprised mRNAs encoding functions for neuronal metabolism and life (19%), nuclear transcription control (15%), cell survival and proliferation (13%), RNA processing and translation (12%). They did not contain any known members of the olfactory transduction pathway. The expression of a sub-set of AOE transcripts was investigated in sub-cellular AOE fractions highly enriched in ciliated dendrites and in AOE fractions after forced hemilateral OSN-specific degeneration. All the mRNAs tested were found to be: i) present in enriched ciliated dendrite preparations ii) down-regulated after OSN degeneration iii) co-purified with polysomal fractions, suggesting their commitment to local translation. We provide strong evidence that the extreme apical side of the olfactory epithelium expresses a unique transcriptome, whose function is not related to olfaction but mainly to defense and survival. The possible local translation of this transcriptome is demonstrated, in supporting cells as well as in olfactory neuron ciliated dendrites., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The olfactory system is regulated by several nervous and hormonal factors, and there is a growing body of evidence that some of these modulations already take place in the olfactory mucosa (OM). We recently suggested that, among others, vasoactive peptides might play multifaceted roles in different OM cells. Here we studied the effect of the vasoconstrictive peptide endothelin (ET) in the rat OM. We identified different components of the ET system both in the olfactory mucosa and in long-term primary culture of OM cells, composed of olfactory sensory neurons (OSNs) lying on a blend of non-neuronal OM cells (nNCs). We demonstrated that ET receptors are differentially expressed on OM cells, and that ET might be locally matured by the endothelin-converting enzyme ECE-1 located in OSNs. Using calcium imaging, we showed that ET triggers robust dose-dependent Ca(2+) responses in most OM cells, which consist of a transient phase, followed, in nNCs, by a sustained plateau phase. All transient responses depended on intracellular calcium release, while the sustained plateau phase also depended on subsequent external calcium entry. Using both pharmacology and spotting lethal (sl/sl) mutant rats, lacking functional ET(B) receptors, we finally demonstrated that these effects of ET are mediated through ET(B) receptors in OSNs and ET(A) receptors in nNCs.The present study therefore identifies endothelin as a potent endogenous modulator of the olfactory mucosa; specific endothelin-mediated Ca(2+) signals may serve distinct signaling functions, and thereby suggest differential functional roles of endothelin in both neuronal and non-neuronal OM cells.

Although odorant-binding proteins (OBP) are one of the most abundant classes of proteins in the mammalian olfactory mucus, they have only recently been ascribed a functional role in the detection of odorants by olfactory neurons. Among the three OBPs described in the rat, OBP-1f is mainly secreted by the lateral nasal glands (LNG) and Bowman's glands, and its expression is transcriptionally regulated by food deprivation in the olfactory mucosa, but not in LNG. Therefore, mucus composition might be locally regulated by hormones or molecules relevant to nutritional status. Our aim has been to investigate the mechanisms of such physiological regulation at the cellular level, through both the examination of OBP-1f synthesis sites in the olfactory mucosa and their putative regulation by leptin, a locally acting satiety hormone. Immunohistochemical observations have allowed the identification of a novel population of OBP-1f-secreting cells displaying morphological and functional characteristics similar to those of epithelial mucous cells. Ultrastructural analyses by both transmission and scanning electron microscopy has enabled a more complete cytoarchitectural characterization of these specialized olfactory mucous cells in their tissue environment. These globular cells are localized in discrete zones of the olfactory epithelium, mainly in the fourth turbinate, and are often scattered from the basal to the apical surface of the epithelium. They contain numerous small droplets of mucosubstances. Using an in-vitro-derived model of olfactory mucosa primary culture, we have been able to demonstrate that leptin increases the production of mucus by these cells, so that they constitute potential targets for the physiological modulation of mucus composition by nutritional cues.

Food odours are major determinants for food choice; their detection is influenced by nutritional status. Among different metabolic signals, insulin plays a major role in food intake regulation. The aim of the present study was to investigate a potential role of insulin in the olfactory mucosa (OM), using ex vivo tissues and in vitro primary cultures. We first established the expression of insulin receptor (IR) in rat olfactory mucosa. Transcripts of IR-A and IR-B isoforms, as well as IRS-1 and IRS-2, were detected in OM extracts. Using immunocytochemistry, IR protein was located in olfactory receptor neurones, sustentacular and basal cells and in endothelium of the lamina propria vessels. Moreover, the insulin binding capacity of OM was quite high compared to that of olfactory bulb or liver. Besides the main pancreatic insulin source, we demonstrated insulin synthesis at a low level in the OM. Interestingly 48 h of fasting, leading to a decreased plasmatic insulin, increased the number of IR in the OM. Local insulin concentration was also enhanced. These data suggest a control of OM insulin system by nutritional status. Finally, an application of insulin on OM, aiming to mimic postprandial insulin increase, reversibly decreased the amplitude of electro-olfactogramme responses to odorants by approximately 30%. These data provide the first evidence that insulin modulates the most peripheral step of odour detection at the olfactory mucosa level.

Neuroanatomical data show that olfactory mucosa (OM) is a possible place for interactions between nutrition and smell. A combination of differential display mRNA analysis together with a macroarray screening was developed to identify transcripts that are differentially expressed in rat OM following food deprivation. Using this method, backed on a stringent statistical analysis, we identified molecules that fell into several Gene Ontology terms including cellular and physiological process, signal transduction, and binding. Among the 15 most differentially expressed molecules, only one was upregulated, but 14 were downregulated in the fasted state among which was, unexpectedly, odorant-binding protein 1F (OBP-1F). Because of its potential relevance to olfactory physiology, we focused our further analysis on OBP-1F using in situ hybridization, quantitative polymerase chain reaction, and western blot analysis. OBP-1F was highlighted in the lateral nasal glands, but its expression (mRNA and protein) did not change following food deprivation. Only the minor fraction of OBP-1F mRNA expressed by the OM itself was downregulated following 48 h fasting. Altogether, our results suggest that the fine transcriptional control of OBP-1F in the OM following food deprivation could be efficient only at the local level, close to its site of secretion to participate in the perireceptor events of the olfactory signal reception.

Leptin is an adipocyte-derived cytokine that regulates body weight mainly via the long form of the leptin receptor (Ob-Rb). Leptin and its receptors are expressed in several tissues, suggesting that leptin might also be effective peripherally. We hypothesized that, as shown in taste cells, leptin and its receptors isoforms (Ob-Rs) could be present in the rat olfactory mucosa (OM). Using RT-PCR, light and electron microscopy immunohistochemistry (ICC), we found that different isoforms of the receptor were expressed in OM and localized in sustentacular cells and in a subpopulation of maturating neurons; in addition, immunoreactivity was also present in differentiated neurons and enriched at the cilia membranes, where the odorants bind to their receptors. Moreover, using RT-PCR, ICC and RIA measurements, we showed that leptin is synthesized locally in the olfactory mucosa. In addition, we demonstrate that fasting causes a significant enhanced transcription of both leptin and Ob-Rs in rat OM by quantitative RT-PCR data. Altogether, these results strongly suggested that leptin, acting as an endocrine or a paracrine factor, could be an important regulator of olfactory function, as a neuromodulator of the olfactory message in cilia of mature olfactory receptors neurons (ORN), but also for the homeostasis of this complex tissue, acting on differentiating neurons and on sustentacular cells.

The olfactory epithelium (OE) is composed of olfactory sensory neurons (OSNs) and sustentacular cells; it lies in the nasal cavity where it is protected by a thin mucus layer. The finely regulated composition of this mucus provides OSN with a suitable ionic environment. To maintain the functional integrity of the epithelium despite permanent physical, chemical and microbial aggressions, both OSNs and surrounding sustentacular cells are continuously renewed from globose basal cells. Moreover, the sense of smell is involved in so numerous behaviours (feeding, reproduction, etc.) that it has to cross-talk with the endocrine and neuroendocrine systems. Thus, besides its sensory function, the olfactory epithelium is thought to undergo a lot of complex regulatory processes. We therefore studied the effects of various neuropeptides on primary cultures of Sprague-Dawley rat olfactory epithelium cells. We found that arginine-vasopressin (AVP) triggered a robust, dose-dependent calcium increase in these cells. The cell response was essentially ascribed to the V1a AVP receptor, whose presence was confirmed by RT-PCR and immunolabelling. In the culture, V1a but not V1b receptors were present, mainly localized in neurons. In the epithelium, both subtypes were found differentially distributed. V1a-R were localized mainly in globose basal cells and at the apical side of the epithelium, in the area of the dendritic knobs of OSNs. V1b-R were strongly associated with Bowman's gland cells and globose basal cells. These localizations suggested potential multifaceted roles of a hormone, AVP, in the olfactory epithelium.