Olfactory Receptor Proteins Research Articles

Odor Biosensors Based on Cell Expressing Olfactory Receptor: Recent Advances

AbstractInspired by the powerful biological olfaction, scientists extracted numerous materials such as olfactory sensory neuron, olfactory receptor (OR) protein, and odorant binding protein from animal olfactory systems, then combined them with transducers to form multiple odor biosensors. These biosensors, despite well inheriting the sensing ability of creatures, have several drawbacks, such as complex preparation process, unstable sensing material characteristics, and high cost. Unlike the biological materials listed above, cell expressing heterologous OR maintains a stable sensing performance after passaging for multiple generations, also its experimental operation is simple, and cost is low. Therefore, odor biosensors based on cell expressing OR have been well developed in recent years. In this review, we first listed several odor biosensors based on cell expressing OR, mainly focusing on fluorescent and electrophysiological measurement methods. Furthermore, we illustrated the techniques to improve the biosensor performance, e. g., wider detection range, longer lifetime, more OR types, and higher quantification efficiency. In addition, we explained the possible prospects such as big sensor array and predicting odor response.

Odorant-binding proteins as recognition elements for smell exploration

The sense of smell in animals enables them to detect and discriminate among many volatile odor molecules in complex environments. This task is accomplished by the family of G-proteins (olfactory receptor proteins, odorant-binding proteins, and chemosensory proteins). Here, we mechanistically elucidate how odors bind to odorant-binding proteins (OBPs) by studying the binding properties of 50 odorant ligands to six OBPs. We confirmed that OBP7 from Ailuropoda melanoleuca and OBP9 from Anopheles gambiae have binding affinities for 16 odors, and further analyzed the structures and protein-ligand interactions by homology modeling and molecular docking. This study found that OBP7 had an affinity for nonanol and camphor, and the docking results indicated that Glu120 was a residue that played a key role in the binding process of OBP7 to odorant ligands. We also explored the binding properties between OBP9 and ligands as well as the binding mechanism, and found that OBP9 bound well to some odorant molecules with repellent efficacy, such as nonanol, camphor, and citral. These findings not only contributed to our understanding of how OBPs were sensitively bound to odorant ligands but also promoted them as suitable targets for applied research in various industrial fields such as pesticides, cosmetics, and pharmaceuticals.

High-throughput ligand profile characterization in novel cell lines expressing seven heterologous insect olfactory receptors for the detection of volatile plant biomarkers

Agriculturally important crop plants emit a multitude of volatile organic compounds (VOCs), which are excellent indicators of their health status and their interactions with pathogens and pests. In this study, we have developed a novel cellular olfactory panel for detecting fungal pathogen-related VOCs we had identified in the field, as well as during controlled inoculations of several crop plants. The olfactory panel consists of seven stable HEK293 cell lines each expressing a functional Drosophila olfactory receptor as a biosensing element along with GCaMP6, a fluorescent calcium indicator protein. An automated 384-well microplate reader was used to characterize the olfactory receptor cell lines for their sensitivity to reference VOCs. Subsequently, we profiled a set of 66 VOCs on all cell lines, covering a concentration range from 1 to 100 μM. Results showed that 49 VOCs (74.2%) elicited a response in at least one olfactory receptor cell line. Some VOCs activated the cell lines even at nanomolar (ppb) concentrations. The interaction profiles obtained here will support the development of biosensors for agricultural applications. Additionally, the olfactory receptor proteins can be purified from these cell lines with sufficient yields for further processing, such as structure determination or integration with sensor devices.

Mutagenesis of the odorant receptor co-receptor (Orco) reveals severe olfactory defects in the crop pest moth Helicoverpa armigera

BackgroundOdorant receptors (ORs) as odorant-gated ion channels play a crucial role in insect olfaction. They are formed by a heteromultimeric complex of the odorant receptor co-receptor (Orco) and a ligand-selective Or. Other types of olfactory receptor proteins, such as ionotropic receptors (IRs) and some gustatory receptors (GRs), are also involved in the olfactory system of insects. Orco as an obligatory subunit of ORs is highly conserved, providing an opportunity to systematically evaluate OR-dependent olfactory responses.ResultsHerein, we successfully established a homozygous mutant (Orco−/−) of Helicoverpa armigera, a notorious crop pest, using the CRISPR/Cas9 gene editing technique. We then compared the olfactory response characteristics of wild type (WT) and Orco−/− adults and larvae. Orco−/− males were infertile, while Orco−/− females were fertile. The lifespan of Orco−/− females was longer than that of WT females. The expressions of most Ors, Irs, and other olfaction-related genes in adult antennae of Orco−/− moths were not obviously affected, but some of them were up- or down-regulated. In addition, there was no change in the neuroanatomical phenotype of Orco−/− moths at the level of the antennal lobe (including the macroglomerular complex region of the male). Using EAG and SSR techniques, we discovered that electrophysiological responses of Orco−/− moths to sex pheromone components and many host plant odorants were absent. The upwind flight behaviors toward sex pheromones of Orco−/− males were severely reduced in a wind tunnel experiment. The oviposition selectivity of Orco−/− females to the host plant (green pepper) has completely disappeared, and the chemotaxis toward green pepper was also lost in Orco−/− larvae.ConclusionsOur study indicates that OR-mediated olfaction is essential for pheromone communication, oviposition selection, and larval chemotaxis of H. armigera, suggesting a strategy in which mate searching and host-seeking behaviors of moth pests could be disrupted by inhibiting or silencing Orco expression.

Olfactory dysfunction in COVID-19: pathology and long-term implications for brain health.

Decreased smell function is related to brain health, future mortality, and quality of life. Most people inflicted with the SARS-CoV-2 virus evidence some measurable smell dysfunction during its acute phase, although many are unaware of the loss. Long-term deficits occur in up to 30% of COVID-19 cases, although total anosmia is relatively rare. This review explores what is presently known about the nature and pathophysiology of olfactory dysfunction due to the SARS-CoV-2 virus, including reversible inflammation within the olfactory cleft, downregulation of olfactory receptor proteins, and long-lasting peripheral and central damage to olfactory structures. It also addresses the question as to whether long-term smell loss might increase the likelihood of future development of cognitive and neurological deficits.

Signatures of conserved and unique molecular features in Afrotheria

Afrotheria is a clade of African-origin species with striking dissimilarities in appearance and habitat. In this study, we compared whole proteome sequences of six Afrotherian species to obtain a broad viewpoint of their underlying molecular make-up, to recognize potentially unique proteomic signatures. We find that 62% of the proteomes studied here, predominantly involved in metabolism, are orthologous, while the number of homologous proteins between individual species is as high as 99.5%. Further, we find that among Afrotheria, L. africana has several orphan proteins with 112 proteins showing < 30% sequence identity with their homologues. Rigorous sequence searches and complementary approaches were employed to annotate 156 uncharacterized protein sequences and 28 species-specific proteins. For 122 proteins we predicted potential functional roles, 43 of which we associated with protein- and nucleic-acid binding roles. Further, we analysed domain content and variations in their combinations within Afrotheria and identified 141 unique functional domain architectures, highlighting proteins with potential for specialized functions. Finally, we discuss the potential relevance of highly represented protein families such as MAGE-B2, olfactory receptor and ribosomal proteins in L. africana and E. edwardii, respectively. Taken together, our study reports the first comparative study of the Afrotherian proteomes and highlights salient molecular features.

Genetic mutation makes fermented fish less stinky

Fermented fish is notorious for its foul odor, but some people can easily stomach the stench of trimethylamine . That may be thanks to a genetic mutation. A genome-wide association study led by Rósa Gísladóttir and Kári Stefánsson from deCODE Genetics turned up a variant of a trace amine olfactory receptor protein called TAAR5. The team asked more than 11,000 Icelanders to smell a sample of trimethylamine and describe its aroma. The reserachers found that people with a single missense mutation in the TAAR5 gene were less likely to associate a waft of trimethylamine with fermented shark or skate—if they could detect the rotten-smelling amine at all ( Curr. Biol. 2020, DOI: 10.1016/j.cub.2020.09.012 ). Though the mutation is found in only a small subset of the population, further analysis shows that the TAAR5 variant is more common in Icelanders than in people from Sweden, Southern Europe, and Africa. Fermented fish

Comparative Evaluation of Sensitivity to Hexanal Between Human and Canine Olfactory Receptors

It is known that a dog possesses much better sense of smell than a human. It has been reported that a dog has advantages (compared to a human) in the number of olfactory receptor (OR) genes, proteins and cells, and also nasal structure. However, a definitive reason for superior sensitivity of the canine olfactory system remains controversial. In this study, we compared sensitivity of human and canine olfactory receptors relative to the same condition. Human OR (hOR2W1) and canine OR (cfOR0312), previously identified to recognize hexanal, were inserted to the pcDNA3 vector. This vector was transfected to HEK293 cells. Hexanal-discriminating ability of ORs was confirmed using Fura-2 AM, a dye which illuminates when binding to calcium ions flowing into cells once olfactory signaling occurs. Consequently, cfOR0312 was more sensitive to hexanal than hOR2W1. Comparison of OR sensitivity would be one of the major factors in clarifying difference of sensitivity between human and canine olfactory systems.

Ectopic expression of olfactory receptors and associated G-protein subunits in the head integument of the amphihaline migratory fish hilsa Tenualosa ilisha.

The chemosensory nature of the tissue from the dorsal surface of the head (also termed sensory pad; SP) of the amphihaline diadromous fish hilsa Tenualosa ilisha was investigated for odorant receptor (OR), olfactory marker protein (OMP) and G-protein subunits (Gαs-olf, Gαq, Gαo, Gαi3) through immunolocalization and immunoblotting techniques. The immunolocalization of OR, OMP and G-protein subunits showed clear expression of these proteins in the tissues of the SP. Robust expressions of these proteins in the SP were detected with immunoblot analysis. The strong expression of these proteins in the SP indicates that the tissues from this area in riverine T. ilisha may play significant role in chemosensing and signalling through ectopic expression of olfactory receptor proteins which are otherwise reported in olfactory organs in vertebrates. Being migratory in nature, ectopic expression of these receptors in T. ilisha probably helps them to prevent damage to epidermal tissues of the SP, or they may also utilize them as a chemo and mechanosensory tool to optimize chemo-communications during migration.

Identification of Missing Proteins in Human Olfactory Epithelial Tissue by Liquid Chromatography-Tandem Mass Spectrometry.

We performed proteomic analyses of human olfactory epithelial tissue to identify missing proteins using liquid chromatography-tandem mass spectrometry. Using a next-generation proteomic pipeline with a < 1.0% false discovery rate at the peptide and protein levels, we identified 3731 proteins, among which five were missing proteins (P0C7M7, P46721, P59826, Q658L1, and Q8N434). We validated the identified missing proteins using the corresponding synthetic peptides. No olfactory receptor (OR) proteins were detected in olfactory tissue, suggesting that detection of ORs would be very difficult. We also identified 49 and 50 alternative splicing variants mapped at the neXtProt and GENCODE databases, respectively, and 2000 additional single amino acid variants. This data set is available at the ProteomeXchange consortium via PRIDE repository (PXD010025).

Expression differences in Aphidius ervi (Hymenoptera: Braconidae) females reared on different aphid host species.

The molecular mechanisms that allow generalist parasitoids to exploit many, often very distinct hosts are practically unknown. The wasp Aphidius ervi, a generalist koinobiont parasitoid of aphids, was introduced from Europe into Chile in the late 1970s to control agriculturally important aphid species. A recent study showed significant differences in host preference and host acceptance (infectivity) depending on the host A. ervi were reared on. In contrast, no genetic differentiation between A. ervi populations parasitizing different aphid species and aphids of the same species reared on different host plants was found in Chile. Additionally, the same study did not find any fitness effects in A. ervi if offspring were reared on a different host as their mothers. Here, we determined the effect of aphid host species (Sitobion avenae versus Acyrthosiphon pisum reared on two different host plants alfalfa and pea) on the transcriptome of adult A. ervi females. We found a large number of differentially expressed genes (between host species: head: 2,765; body: 1,216; within the same aphid host species reared on different host plants: alfalfa versus pea: head 593; body 222). As expected, the transcriptomes from parasitoids reared on the same host species (pea aphid) but originating from different host plants (pea versus alfalfa) were more similar to each other than the transcriptomes of parasitoids reared on a different aphid host and host plant (head: 648 and 1,524 transcripts; body: 566 and 428 transcripts). We found several differentially expressed odorant binding proteins and olfactory receptor proteins in particular, when we compared parasitoids from different host species. Additionally, we found differentially expressed genes involved in neuronal growth and development as well as signaling pathways. These results point towards a significant rewiring of the transcriptome of A. ervi depending on aphid-plant complex where parasitoids develop, even if different biotypes of a certain aphid host species (A. pisum) are reared on the same host plant. This difference seems to persist even after the different wasp populations were reared on the same aphid host in the laboratory for more than 50 generations. This indicates that either the imprinting process is very persistent or there is enough genetic/allelic variation between A. ervi populations. The role of distinct molecular mechanisms is discussed in terms of the formation of host fidelity.

Analysis of Differentially Expressed Genes in Gastrocnemius Muscle between DGAT1 Transgenic Mice and Wild-Type Mice

Adipose tissue was the major energy deposition site of the mammals and provided the energy for the body and released the external pressure to the internal organs. In animal production, fat deposition in muscle can affect the meat quality, especially the intramuscular fat (IMF) content. Diacylglycerol acyltransferase-1 (DGAT1) was the key enzyme to control the synthesis of the triacylglycerol in adipose tissue. In order to better understand the regulation mechanism of the DGAT1 in the intramuscular fat deposition, the global gene expression profiling was performed in gastrocnemius muscle between DGAT1 transgenic mice and wild-type mice by microarray. 281 differentially expressed transcripts were identified with at least 1.5-fold change and the p value < 0.05. 169 transcripts were upregulated and 112 transcripts were downregulated. Ten genes (SREBF1, DUSP1, PLAGL1, FKBP5, ZBTB16, PPP1R3C, CDC14A, GLUL, PDK4, and UCP3) were selected to validate the reliability of the chip's results by the real-time PCR. The finding of RT-PCR was consistent with the gene chip. Seventeen signal pathways were analyzed using KEGG pathway database and the pathways concentrated mainly on the G-protein coupled receptor protein signaling pathway, signal transduction, oxidation-reduction reaction, olfactory receptor activity, protein binding, and zinc ion binding. This study implied a function role of DGAT1 in the synthesis of TAG, insulin resistance, and IMF deposition.

Novel Paradigm for Odorant-Olfactory Receptor Interactions

This opinion paper recommends a novel methodology to mechanistically elucidate the interactions between olfactory receptors and odorant molecules. Preliminary results presented here show diffusion of odorant-ligand molecules from outside the olfactory receptor protein into the receptor binding region. This notion marks a sharp contrast with computational studies related to olfactory receptor-odorant interactions, which have drawn conclusions from static or dynamic studies that have started from the ligand already docked in a preferred configuration in the receptor’s binding region. Dynamic simulations studies that begin by tracking the approach of the odorant from outside the protein and its transit to the receptor binding region followed by tracking the rest of the receptor-odorant interactions will allow the determination of protein structural changes: 1) that facilitate the odorant path; 2) that facilitate specific interactions between amino acids and the odorant; 3) that identify the activation state of the protein and its transition to the inactive state. The novel methodology will not only improve comparisons of hypotheses generated from these computational studies with experimental studies, it will also provide novel mechanistic insights into these interactions. The primary goal is to establish a predictive paradigm for olfactory receptor-odorant interactions.

Immunochemical strategy for quantification of G-coupled olfactory receptor proteins on natural nanovesicles

Cell membrane proteins are involved in a variety of biochemical pathways and therefore constitute important targets for therapy and development of new drugs. Bioanalytical platforms and binding assays using these membrane protein receptors for drug screening or diagnostic require the construction of well-characterized liposome and lipid bilayer arrays that act as support to prevent protein denaturation during biochip processing. Quantification of the protein receptors in the lipid membrane arrays is a key issue in order to produce reproducible and well-characterized chips. Herein, we report a novel immunochemical analytical approach for the quantification of membrane proteins (i.e., G-protein-coupled receptor, GPCR) in nanovesicles (NVs). The procedure allows direct determination of tagged receptors (i.e., c-myc tag) without any previous protein purification or extraction steps. The immunochemical method is based on a microplate ELISA format and quantifies this tag on proteins embedded in NVs with detectability in the picomolar range, using protein bioconjugates as reference standards. The applicability of the method is demonstrated through the quantification of the c-myc-olfactory receptor (OR, c-myc-OR1740) in the cell membrane NVs. The reported method opens the possibility to develop well-characterized drug-screening platforms based on G-coupled proteins embedded on membranes.

Sequestration within nuclear chromocenters is not a requirement for silencing olfactory receptor transcription in a placode-derived cell line

Mouse olfaction depends on specialized olfactory sensory neurons (OSNs) that each express only one olfactory receptor protein from among a family of >1000 olfactory receptor (OR) genes encoded in the genome. To investigate epigenetic mechanisms underlying monogenic OR expression, we characterized the nuclear organization of OR loci in an olfactory placode-derived cell line (OP6) derived from a pre-neuronal cell along the OSN lineage. OR loci are significantly enriched within nuclear chromocenters in these cells as compared with control loci tested. However, we observe variability in chromocenter occupancy among different OR loci and from cell-to-cell, suggesting that these associations are transient or context dependent. The lamin B receptor (LBR), whose downregulation is necessary for aggregation of chromocenters and OR genes in mature OSNs, exhibits an unusual non-peripheral expression pattern in OP6 nuclei; upon further OP6 cell differentiation, LBR expression is lost and chromocenters begin to aggregate. However, neither undifferentiated nor differentiated OP6 cells sequester OR genes within the chromocenters, despite the establishment of monogenic OR expression in these cells. These results indicate that sequestration of competing OR loci is not a requirement for monogenic OR expression in OP6 cells, and could indicate that the initial establishment of monogenic OR expression during OSN differentiation in vivo occurs prior to recruitment of OR genes into chromocenters.

Narrow tuning of an odorant receptor to plant volatiles in Spodoptera exigua (Hübner).

Olfaction plays an important role in insects in recognizing volatile compounds, which are used to find food and mates, as well as avoid danger, predators and pathogens. The key players in the detection of volatile compounds are olfactory receptor (OR) proteins, which are located within the dendritic membrane of sensory neurons and extend into the lymph of sensilla on insect antennae. In the present study, we identify an OR gene, named SexiOR3, which is expressed in adult antenna in both sexes, but with female bias, in the beet armyworm moth Spodoptera exigua. Further in situ hybridization analysis revealed that SexiOR3 was mainly located in short trichoid sensilla. In in vitro heterologous expression experiments, SexiOR3 was narrowly tuned to E-β-farnesene and several compounds of related structure, among 62 different compounds tested in this study. Furthermore, SexiOR3 responds to E-β-farnesene at a low concentration of 10(-9) M, comparable to that of pheromone receptors (PRs) responding to the pheromones. This is a very interesting finding for a general OR, indicating that high specificity is a feature of at least one general OR and not only the PRs. The results suggest that the OR3 gene may play an important role in the moth olfactory system, and underpins important insect behaviour.

The membrane proteome of sensory cilia to the depth of olfactory receptors.

In the nasal cavity, the nonmotile cilium of olfactory sensory neurons (OSNs) constitutes the chemosensory interface between the ambient environment and the brain. The unique sensory organelle facilitates odor detection for which it includes all necessary components of initial and downstream olfactory signal transduction. In addition to its function in olfaction, a more universal role in modulating different signaling pathways is implicated, for example, in neurogenesis, apoptosis, and neural regeneration. To further extend our knowledge about this multifunctional signaling organelle, it is of high importance to establish a most detailed proteome map of the ciliary membrane compartment down to the level of transmembrane receptors. We detached cilia from mouse olfactory epithelia via Ca(2+)/K(+) shock followed by the enrichment of ciliary membrane proteins at alkaline pH, and we identified a total of 4,403 proteins by gel-based and gel-free methods in conjunction with high resolution LC/MS. This study is the first to report the detection of 62 native olfactory receptor proteins and to provide evidence for their heterogeneous expression at the protein level. Quantitative data evaluation revealed four ciliary membrane-associated candidate proteins (the annexins ANXA1, ANXA2, ANXA5, and S100A5) with a suggested function in the regulation of olfactory signal transduction, and their presence in ciliary structures was confirmed by immunohistochemistry. Moreover, we corroborated the ciliary localization of the potassium-dependent Na(+)/Ca(2+) exchanger (NCKX) 4 and the plasma membrane Ca(2+)-ATPase 1 (PMCA1) involved in olfactory signal termination, and we detected for the first time NCKX2 in olfactory cilia. Through comparison with transcriptome data specific for mature, ciliated OSNs, we finally delineated the membrane ciliome of OSNs. The membrane proteome of olfactory cilia established here is the most complete today, thus allowing us to pave new avenues for the study of diverse molecular functions and signaling pathways in and out of olfactory cilia and thus to advance our understanding of the biology of sensory organelles in general.

Learning olfactory codes using matrix factorization on 2DG uptake patterns from rats

Recent studies based on both the analysis of odor descriptors [1] and psychophysical studies [2] have suggested that olfactory perception is mainly dominated by odor pleasantness. In contrast, findings on the sensory side [3] as well as on the perceptual side [4] indicate an odor perception space that is spanned by about 30 dimensions. On the way to understand the basic principles in the decoding of olfactory information from the single molecular compounds, we are focusing in this study on the first level of information processing in the sense of smell: The olfactory bulb (OB). We know that the olfactory epithelium is separated into four regions [5] of olfactory receptor protein expression and we expect a similar organization in the olfactory bulb [6]. With 2-DG-uptake-images, Johnson et al. [7] measured odorant-specific activity patterns, covering 98% of the bulbar surface with a resolution of one glomerulus per pixel. Using a database of 143 such bulbar activity fingerprints, we propose the so called non-negative matrix factorization [8] to decompose the data into a set of common features. These features can be best described as the coding alphabet while the uptake images of single molecular compounds resemble natural occurring code words. Interestingly, we can achieve three meaningful alphabets with a different size: 4, 6, and 9 digits. Only the last one is suitable for actually coding the compounds, as a single compound is evoking activity of only about 1/8 of the bulbar surface, with an average code word length of two to three digits. The other two decompositions might reflect the superglomerular organization of the receptor neurons in the bulb, like e.g. the above mentioned expression patterns and neuronal subnetworks that directly result from the targeting mechanisms guiding the neurons from the OE to the OB. We argue that the proposed decompositions are not only reflecting the chemotopic organization of the bulbus but rather giving a first hint on the coding scheme underlying olfactory perception.

Genetic basis of olfactory cognition: extremely high level of DNA sequence polymorphism in promoter regions of the human olfactory receptor genes revealed using the 1000 Genomes Project dataset

The molecular mechanism of olfactory cognition is very complicated. Olfactory cognition is initiated by olfactory receptor proteins (odorant receptors), which are activated by olfactory stimuli (ligands). Olfactory receptors are the initial player in the signal transduction cascade producing a nerve impulse, which is transmitted to the brain. The sensitivity to a particular ligand depends on the expression level of multiple proteins involved in the process of olfactory cognition: olfactory receptor proteins, proteins that participate in signal transduction cascade, etc. The expression level of each gene is controlled by its regulatory regions, and especially, by the promoter [a region of DNA about 100–1000 base pairs long located upstream of the transcription start site (TSS)]. We analyzed single nucleotide polymorphisms using human whole-genome data from the 1000 Genomes Project and revealed an extremely high level of single nucleotide polymorphisms in promoter regions of olfactory receptor genes and HLA genes. We hypothesized that the high level of polymorphisms in olfactory receptor promoters was responsible for the diversity in regulatory mechanisms controlling the expression levels of olfactory receptor proteins. Such diversity of regulatory mechanisms may cause the great variability of olfactory cognition of numerous environmental olfactory stimuli perceived by human beings (air pollutants, human body odors, odors in culinary etc.). In turn, this variability may provide a wide range of emotional and behavioral reactions related to the vast variety of olfactory stimuli.

Two distinct types of neuronal asymmetries are controlled by the Caenorhabditis elegans zinc finger transcription factor die-1

Left/right asymmetric features of animals are either randomly distributed on either the left or right side within a population ("antisymmetries") or found stereotypically on one particular side of an animal ("directional asymmetries"). Both types of asymmetries can be found in nervous systems, but whether the regulatory programs that establish these asymmetries share any mechanistic features is not known. We describe here an unprecedented molecular link between these two types of asymmetries in Caenorhabditis elegans. The zinc finger transcription factor die-1 is expressed in a directionally asymmetric manner in the gustatory neuron pair ASE left (ASEL) and ASE right (ASER), while it is expressed in an antisymmetric manner in the olfactory neuron pair AWC left (AWCL) and AWC right (AWCR). Asymmetric die-1 expression is controlled in a fundamentally distinct manner in these two neuron pairs. Importantly, asymmetric die-1 expression controls the directionally asymmetric expression of gustatory receptor proteins in the ASE neurons and the antisymmetric expression of olfactory receptor proteins in the AWC neurons. These asymmetries serve to increase the ability of the animal to discriminate distinct chemosensory inputs.