Olfactory Desensitization Research Articles

Optogenetic Manipulation of Olfactory Responses in Transgenic Zebrafish: A Neurobiological and Behavioral Study.

Olfaction is an important neural system for survival and fundamental behaviors such as predator avoidance, food finding, memory formation, reproduction, and social communication. However, the neural circuits and pathways associated with the olfactory system in various behaviors are not fully understood. Recent advances in optogenetics, high-resolution in vivo imaging, and reconstructions of neuronal circuits have created new opportunities to understand such neural circuits. Here, we generated a transgenic zebrafish to manipulate olfactory signal optically, expressing the Channelrhodopsin (ChR2) under the control of the olfactory specific promoter, omp. We observed light-induced neuronal activity of olfactory system in the transgenic fish by examining c-fos expression, and a calcium indicator suggesting that blue light stimulation caused activation of olfactory neurons in a non-invasive manner. To examine whether the photo-activation of olfactory sensory neurons affect behavior of zebrafish larvae, we devised a behavioral choice paradigm and tested how zebrafish larvae choose between two conflicting sensory cues, an aversive odor or the naturally preferred phototaxis. We found that when the conflicting cues (the preferred light and aversive odor) were presented together simultaneously, zebrafish larvae swam away from the aversive odor. However, the transgenic fish with photo-activation were insensitive to the aversive odor and exhibited olfactory desensitization upon optical stimulation of ChR2. These results show that an aversive olfactory stimulus can override phototaxis, and that olfaction is important in decision making in zebrafish. This new transgenic model will be useful for the analysis of olfaction related behaviors and for the dissection of underlying neural circuits.

Attentional modulation of desensitization to odor

Subjective and behavioral responsiveness to odor diminishes during prolonged exposure. The precise mechanisms underlying olfactory desensitization are not fully understood, but previous studies indicate that the phenomenon may be modulated by central-cognitive processes. The present study investigated the effect of attention on perceived intensity during exposure to a pleasant odor. A within-subjects design was utilized with 19 participants attending 2 sessions. During each session, participants continuously rated their perceived intensity of a 10-minute exposure to a pleasant fragrance administered using an olfactometer. An auditory oddball task was implemented to manipulate the focus of attention in each session. Participants were instructed to either direct their attention toward the sounds, but still to rate odor, or to focus entirely on rating the odor. Analysis revealed three 50-second time windows with significantly lower mean intensity ratings during the distraction condition. Curve fitting of the data disclosed a linear function of desensitization in the focused attention condition compared with an exponential decay function during distraction condition, indicating an increased rate of initial desensitization when attention is distracted away from the odor. In the focused-attention condition, perceived intensity demonstrated a regular pattern of odor sensitivity occurring at approximately 1−2 minutes intervals following initial desensitization. Spectral analysis of low-frequency oscillations confirmed the presence of augmented spectral power in this frequency range during focused relative to distracted conditions. The findings demonstrate for the first time modulation of odor desensitization specifically by attentional factors, exemplifying the relevance of top-down control for ongoing perception of odor.

Subjective Olfactory Desensitization and Recovery in Humans

Adaptation to smells is a well-known phenomenon and appears to be one of the major characteristics of olfaction. However, no standardized protocols for the psychophysical measurement of olfactory adaptation and recovery are available to date. Twenty normosmic participants were included. Hydrogen sulfide (H2S) and phenylethyl alcohol were used in different concentrations based on air dilution olfactometry. Volunteers were exposed to a constant flow of odorous air until perception disappeared completely. For testing recovery, the volunteers were exposed to a reference stimulus following complete self-desensitization with the same odor. The subjects were then again exposed to the same odorant after recovery periods of different lengths and instructed to rate stimulus intensity. The time to complete desensitization increased with increasing stimulus concentration for both odorants. Subjects desensitized more rapidly using H2S. Olfactory adaptation led to a reduction in stimulus intensity for the subsequent identical stimulation. Longer recovery periods resulted in increased intensity of the subsequent stimulus independent of the stimulus used. The results confirm current knowledge regarding the dynamics of olfactory adaptation and demonstrate differences in olfactory desensitization between the 2 odorants used. Olfactory recovery was independent of the odorant used, indicating that olfactory recovery after complete desensitization may be a uniform process.

Odorant-specific requirements for arrestin function inDrosophilaolfaction

The ability to modulate olfactory sensitivity is necessary to detect chemical gradients and discriminate among a multitude of odor stimuli. Desensitization of odorant receptors has been postulated to occur when arrestins prevent the activation of downstream second messengers. A paucity of in vivo data on olfactory desensitization prompts use of Drosophila melanogaster genetics to investigate arrestins' role in regulating olfactory signaling pathways. Physiological analysis of peripheral olfactory sensitivity reveals decreased responsiveness to a host of chemically distinct odorants in flies deficient for arrestin1 (arr1), arrestin2 (arr2), or both. These phenotypes are manifest in odorant- and dose- dependent fashions. Additionally, mutants display altered adaptive properties under a prolonged exposure paradigm. Behaviorally, arr1 mutants are impaired in olfactory-based orientation towards attractive odor sources. As the olfactory deficits vary according to chemical identity and concentration, they indicate that a spectrum of arrestin activity is essential for odor processing depending upon the particular olfactory pathway involved. Arrestin mutant phenotypes are hypothesized to be a consequence of down-regulation of olfactory signaling to avoid cellular excitotoxicity. Importantly, phenotypic rescue of olfactory defects in arr1(1) mutants is achieved through transgenic expression of wild-type arr1. Taken together, these data clearly indicate that arrestins are required in a stimulus-specific manner for wild type olfactory function and add another level of complexity to peripheral odor coding mechanisms that ultimately impact olfactory behavior.

Recording of the human electro-olfactogram

Electro-olfactograms (EOG) are electrical potentials of the olfactory epithelium that occur in response to olfactory stimulation. The EOG represents the sum of generator potentials of olfactory receptor neurons. While this response has been used extensively in animal research, there are only a handful of papers describing the properties of the human EOG. In addition to a discussion of methodological issues related to the EOG, this review summarizes the characteristics and uses of these recordings. Among other results, EOGs have been used to provide evidence for the dominant role of the central nervous system in olfactory desensitization, for the functional characterization of the olfactory epithelium, the specific topographical distribution of olfactory receptors, or the expression of olfactory receptor neurons in response to exposure to odorants, and the characterization of certain odorants as olfactory receptor antagonists. In conclusion, in combination with nasal endoscopy and air-dilution olfactometry, the EOG is a unique part of a large array of techniques used to provide a complete picture of the processing of olfactory information in humans.

Recording of the human electro-olfactogram

Electro-olfactograms (EOG) are electrical potentials of the olfactory epithelium that occur in response to olfactory stimulation. The EOG represents the sum of generator potentials of olfactory receptor neurons. While this response has been used extensively in animal research, there are only a handful of papers describing the properties of the human EOG. In addition to a discussion of methodological issues related to the EOG, this review summarizes the characteristics and uses of these recordings. Among other results, EOGs have been used to provide evidence for the dominant role of the central nervous system in olfactory desensitization, for the functional characterization of the olfactory epithelium, the specific topographical distribution of olfactory receptors, or the expression of olfactory receptor neurons in response to exposure to odorants, and the characterization of certain odorants as olfactory receptor antagonists. In conclusion, in combination with nasal endoscopy and air-dilution olfactometry, the EOG is a unique part of a large array of techniques used to provide a complete picture of the processing of olfactory information in humans.

Odorant Receptors and Desensitization Proteins Colocalize in Mammalian Sperm

The identification of transcripts encoding putative olfactory receptors in mammalian germ cells (1) has generated the hypothesis that olfactory receptors may serve a chemosensory role in sperm chemotaxis during fertilization. We have sought to identify and localize these receptors and their regulatory machinery in rat sperm in order to gain further insight into mammalian sperm chemotaxis and odorant receptor physiology. We conducted reverse transcription-polymerase chain reaction (RT-PCR) using degenerate primers directed against sequences conserved across members of the known odorant receptor family to identify transcripts from testis and round spermatids. Western analysis and immunohistochemistry were performed using antibodies raised against two peptide sequences conserved among odorant receptors and using fusion protein antibodies to G-protein receptor kinase 3 (GRK3/beta ARK2) and beta-arrestin2. We detected transcripts encoding putative odorant receptors in both testis and round spermatids of the adult rat. Restriction digests of the PCR products demonstrated the existence of multiple gene products. Two anti-odorant receptor antibodies specifically recognized a 64 kD band in rat sperm preparations by Western blot. The proteins GRK3 and beta-arrestin2, implicated in olfactory desensitization, were detected in sperm cytosolic extracts using Western analysis. Immunohistochemistry colocalized putative odorant receptors, GRK3 and beta-arrestin2 to elongating spermatids in the testis and to the midpiece of mature sperm. The specific localization of odorant receptors to the respiratory center of mature sperm is consistent with a role for these proteins in transducing chemotactic signals. Based on the colocalization, it is plausible that GRK3 and beta-arrestin2 function in sperm to regulate putative chemoreceptor responses.

Olfactory desensitization requires membrane targeting of receptor kinase mediated by beta gamma-subunits of heterotrimeric G proteins.

Olfaction is mediated by G protein-coupled receptors. In isolated rat olfactory cilia, odorants such as citralva stimulate a burst of cAMP, which peaks in 50 ms and returns almost to base-line level within 150 ms in the continuing presence of odorant. This desensitization is mediated by the cAMP dependent protein kinase and a specialized G protein-coupled receptor kinase originally termed beta ARK2 (GRK3). In vitro experiments suggest that the prenylated beta gamma-subunits of heterotrimeric G proteins target the cytosolic beta ARK1 (GRK2) enzyme to its membrane bound receptor substrate by binding to sites in its carboxyl terminus. Here we demonstrate that odorants stimulate translocation of GRK3 from cytosol to membranes in isolated rat olfactory cilia. We introduced a glutathione S-transferase-GRK3ct fusion protein, containing the carboxyl-terminal 222 amino acid residues of GRK3, which includes the beta gamma binding site, or a 28-amino acid peptide derived therefrom, into permeabilized cilia preparations. These reagents block odorant-mediated enzyme translocation and desensitization while markedly attenuating odorant-stimulated phosphorylation of olfactory proteins. These findings suggest that beta gamma-subunits may physiologically regulate a G protein-coupled receptor kinase and that enzyme translocation may be a general and required feature of the activity of some members of this enzyme family.

A beta-adrenergic receptor kinase-like enzyme is involved in olfactory signal termination.

We have previously shown that second-messenger-dependent kinases (cAMP-dependent kinase, protein kinase C) in the olfactory system are essential in terminating second-messenger signaling in response to odorants. We now document that subtype 2 of the beta-adrenergic receptor kinase (beta ARK) is also involved in this process. By using subtype-specific antibodies to beta ARK-1 and beta ARK-2, we show that beta ARK-2 is preferentially expressed in the olfactory epithelium in contrast to findings in most other tissues. Heparin, an inhibitor of beta ARK, as well as anti-beta ARK-2 antibodies, (i) completely prevents the rapid decline of second-messenger signals (desensitization) that follows odorant stimulation and (ii) strongly inhibits odorant-induced phosphorylation of olfactory ciliary proteins. In contrast, beta ARK-1 antibodies are without effect. Inhibitors of protein kinase A and protein kinase C also block odorant-induced desensitization and phosphorylation. These data suggest that a sequential interplay of second-messenger-dependent and receptor-specific kinases is functionally involved in olfactory desensitization.

Learning in a carabid beetle ( Pterostichus melanarius)

This study reports on learning in a predacious ground beetle ( Prerostichus melanarius). The first observations showed an apparent inability to learn spatial information in a maze but indicated the possibility that the beetles could learn to reduce a strong thigmotactic response in an open-field and hence locate a centrally placed water source. The second experiment confirmed the presence of open-field learning and showed that this information is only retained for a period of between 100 and 200 min. Control groups supported the presence of learning but showed that it was only manifested when cued by odour. The final experiment showed that the decline of the effects of the learning experience was not due to a gradual olfactory desensitization to the relevant odour, and that it presumably is the result of actual memory loss.