SHORT COMMUNICATION: Olfactory Receptor Database (ORDB): A Resource for Sharing and Analyzing Published and Unpublished Data

An olfactory receptor database (ORDB) is being developed tofacilitate analysis of this large gene family. ORDB currentlycontains over 400 olfactory receptor sequences and related information,and is available via the World Wide Web. We plan to incorporatefunctional data, structural models, spatial localization andother categories of information, toward an integrated modelof olfactory receptor function. Chem. Senses 22: 321–326,1997.     

Olfactory Receptor Database: a database of the largest eukaryotic gene family.

The Olfactory Receptor Database (ORDB) is a WWW-accessible database that stores data on Olfactory Receptor-like molecules (ORs) and has been open to the public since June 1996. It contains a public and a private area. The public area includes published DNA and protein sequence data for ORs, links to OR models and data on their expression, chromosomal localization and source organism, as well as (i) links to bibliography through PubMed and (ii) interactive WWW-based tools, such as BLAST homology searching. The private area functions as a service to laboratories that are actively cloning receptors. Source laboratories enter the sequences of the receptor clones they have characterized to the private database and can search for identical or near identical OR sequences in both public and private databases. If another laboratory has cloned and deposited an identical or closely matching sequence there are means for communication between the laboratories to help avoid duplication of work. ORDB is available via the WWW at http://crepe.med.yale.edu/ORDB/HTML     

RNA-Seq Analysis of Human Trigeminal and Dorsal Root Ganglia with a Focus on Chemoreceptors

The chemosensory capacity of the somatosensory system relies on the appropriate expression of chemoreceptors, which detect chemical stimuli and transduce sensory information into cellular signals. Knowledge of the complete repertoire of the chemoreceptors expressed in human sensory ganglia is lacking. This study employed the next-generation sequencing technique (RNA-Seq) to conduct the first expression analysis of human trigeminal ganglia (TG) and dorsal root ganglia (DRG). We analyzed the data with a focus on G-protein coupled receptors (GPCRs) and ion channels, which are (potentially) involved in chemosensation by somatosensory neurons in the human TG and DRG. For years, transient receptor potential (TRP) channels have been considered the main group of receptors for chemosensation in the trigeminal system. Interestingly, we could show that sensory ganglia also express a panel of different olfactory receptors (ORs) with putative chemosensory function. To characterize OR expression in more detail, we performed microarray, semi-quantitative RT-PCR experiments, and immunohistochemical staining. Additionally, we analyzed the expression data to identify further known or putative classes of chemoreceptors in the human TG and DRG. Our results give an overview of the major classes of chemoreceptors expressed in the human TG and DRG and provide the basis for a broader understanding of the reception of chemical cues.     

Evolution of odorant receptors

Odorant receptors (ORs) located in the nasal epithelium, at the ciliated surface of olfactory sensory neurons, represent the initial step of a transduction cascade that leads to odor detection. ORs form the largest and most diverse family of G-protein-coupled receptors (GPCRs). They are encoded by a multigene family that has been partially characterized in cyclostomes, teleosts, amphibia, birds and mammals, as well as in Drosophila melanogaster and the nematode Caenorhabditis elegans. As new sequence data emerge, it is increasingly clear that OR primary structure can vary dramatically across phyla. Some chemoreceptors are encoded by genes with little sequence similarity to the prototypical ORs originally isolated in mammals. A large number of sequences are now available allowing a detailed study of the evolutionary implications of OR diversity across species. This review discusses the evolutionary implications of the divergent primary structures of chemoreceptors with identical functions.     

Combining dissimilar senses: central processing of hydrodynamic and chemosensory inputs in aquatic crustaceans

Aquatic environments are by their nature dynamic and dominated by fluid movements driven by lunar tides, temperature and salinity density gradients, wind-driven currents, and currents generated by the earth's rotation. Accordingly, animals within the aquatic realm must be able to sense and respond to both large-scale (advection) and small-scale (eddy turbulence) fluid dynamics, for chemical signals critically important for their survival are embedded within such movements. Aquatic crustaceans possess many types of near-field fluid-flow detectors and two general classes of chemoreceptors on their body appendages: high-threshold, near-field receptors that may be somewhat equated with the sense of taste, and low-threshold far-field receptors that can be considered as olfactory. This review briefly summarizes the distribution of hydrodynamic and high-threshold chemoreceptors in aquatic crustaceans and the physiological characteristics of olfactory receptors in lobsters; it also examines recent physiological evidence for the central nervous integration of inputs from olfactory receptors and hydrodynamic detectors, two dissimilar senses that must be combined within the brain for survival. Marine crustaceans have provided valuable insights about mechanisms of primary olfactory sensory physiology; their additional sensitivity to hydrodynamic stimulation makes them a potentially useful model for examining how these two critical sensory inputs are combined within the brain to enhance foraging behavior. Multimodal sensory processing is critically important to all animals, and the principles and concepts derived from these crustacean studies may provide generalities about neuronal processing across taxa.     

Integrated olfactory receptor and microarray gene expression databases

Background  

Gene expression patterns of olfactory receptors (ORs) are an important component of the signal encoding mechanism in the olfactory system since they determine the interactions between odorant ligands and sensory neurons. We have developed the Olfactory Receptor Microarray Database (ORMD) to house OR gene expression data. ORMD is integrated with the Olfactory Receptor Database (ORDB), which is a key repository of OR gene information. Both databases aim to aid experimental research related to olfaction.     

Distinct evolutionary patterns between chemoreceptors of 2 vertebrate olfactory systems and the differential tuning hypothesis

Most tetrapod vertebrates have 2 olfactory systems, the main olfactory system (MOS) and the vomeronasal system (VNS). According to the dual olfactory hypothesis, the MOS detects environmental odorants, whereas the VNS recognizes intraspecific pheromonal cues. However, this strict functional distinction has been blurred by recent reports that both systems can perceive both types of signals. Studies of a limited number of receptors suggest that MOS receptors are broadly tuned generalists, whereas VNS receptors are narrowly tuned specialists. However, whether this distinction applies to all MOS and VNS receptors remains unknown. The differential tuning hypothesis predicts that generalist MOS receptors detect an overlapping set of ligands and thus are more likely to be conserved over evolutionary time than specialist VNS receptors, which would evolve in a more lineage-specific manner. Here we test this prediction for all olfactory chemoreceptors by examining the evolutionary patterns of MOS-expressed odorant receptors (ORs) and trace amine-associated receptors (TAARs) and VNS-expressed vomeronasal type 1 receptors (V1Rs) and vomeronasal type 2 receptors (V2Rs) in 7 tetrapods (mouse, rat, dog, opossum, platypus, chicken, and frog). The phylogenies of V1Rs and V2Rs show abundant lineage-specific gene gains/losses and virtually no one-to-one orthologs between species. Opposite patterns are found for ORs and TAARs. Analysis of functional data and ligand-binding sites of ORs confirms that paralogous chemoreceptors are more likely than orthologs to have different ligands and that functional divergence between paralogous chemoreceptors is established relatively quickly following gene duplication. Together, these results strongly suggest that the functional profile of the VNS chemoreceptor repertoire evolves much faster than that of the MOS chemoreceptor repertoire and that the differential tuning hypothesis applies to the majority, if not all, of MOS and VNS receptors.     

Fine structure of tarsal sensory organs in the whip spider Admetus pumilio (Amblypygi, Arachnida).

The sensory organs on the tarsi of the antenniform first legs of the whip spider Admetus pumilio C. L. Koch (Amblypygi, Arachnida) were examined with the scanning and transmission electron microscope. At least four different types of hair sensilla were found: (1) thick-walled bristles, which have the characteristics of contact chemoreceptors (several chemoreceptive dendrites in the lumen plus two mechanoreceptors at the base); (2) short club sensilla, innervated by 4-6 neurons which terminate in a pore on the tip; they are possibly humidity receptors; (3) porous sensilla, which are either innervated by 20-25 neurons and have typical pore tubules, or they have 40-45 neurons but no pore tubules; both types are considered to be olfactory; (4) rod sensilla occur in clusters near segmental borders; they are innervated by only one large dendrite which branches inside the lumen. Other tarsal receptors are the claws, which correspond to contact chemoreceptors, and the pit organ which resembles the tarsal organ of spiders. Compared to other arthropod sensilla, the contact chemoreceptors are very similar to those of spiders, while the porous sensilla correspond structurally to olfactory receptors in insects; the club and rod sensilla seem to be typical for amblypygids.     

SenseLab: new developments in disseminating neuroscience information

This article presents the latest developments in neuroscience information dissemination through the SenseLab suite of databases: NeuronDB, CellPropDB, ORDB, OdorDB, OdorMapDB, ModelDB and BrainPharm. These databases include information related to: (i) neuronal membrane properties and neuronal models, and (ii) genetics, genomics, proteomics and imaging studies of the olfactory system. We describe here: the new features for each database, the evolution of SenseLab's unifying database architecture and instances of SenseLab database interoperation with other neuroscience online resources.     

Roles of odorant receptors in projecting axons in the mouse olfactory system

In the mouse olfactory epithelium, there are about ten million olfactory sensory neurons, each expressing a single type of odorant receptor out of approximately 1000. Olfactory sensory neurons expressing the same odorant receptor converge their axons to a specific set of glomeruli on the olfactory bulb. How odorant receptors play an instructive role in the projection of axons to the olfactory bulb has been one of the major issues of developmental neurobiology. Recent studies revealed previously overlooked roles of odorant receptor-derived cAMP signals in the axonal projection of olfactory sensory neurons; the levels of cAMP and neuronal activity appear to determine the expression levels of axon guidance/sorting molecules and thereby direct the axonal projection of olfactory sensory neurons. These findings provide new insights as to how peripheral inputs instruct neuronal circuit formation in the mammalian brain.     

The mysterious multi-modal repellency of DEET

DEET is the most effective insect repellent available and has been widely used for more than half a century. Here, I review what is known about the olfactory and contact mechanisms of DEET repellency. For mosquitoes, DEET has at least two molecular targets: Odorant Receptors (ORs) mediate the effect of DEET at a distance, while unknown chemoreceptors mediate repellency upon contact. Additionally, the ionotropic receptor Ir40a has recently been identified as a putative DEET chemosensor in Drosophila. The mechanism of how DEET manipulates these molecular targets to induce insect avoidance in the vapor phase is also contested. Two hypotheses are the most likely: DEET activates an innate olfactory neural circuit leading to avoidance of hosts (smell and avoid hypothesis) or DEET has no behavioral effect on its own, but instead acts cooperatively with host odors to drive repellency (confusant hypothesis). Resolving this mystery will inform the search for a new generation of insect repellents.     

The mysterious multi-modal repellency of DEET

DEET is the most effective insect repellent available and has been widely used for more than half a century. Here, I review what is known about the olfactory and contact mechanisms of DEET repellency. For mosquitoes, DEET has at least two molecular targets: Odorant Receptors (ORs) mediate the effect of DEET at a distance, while unknown chemoreceptors mediate repellency upon contact. Additionally, the ionotropic receptor Ir40a has recently been identified as a putative DEET chemosensor in Drosophila. The mechanism of how DEET manipulates these molecular targets to induce insect avoidance in the vapor phase is also contested. Two hypotheses are the most likely: DEET activates an innate olfactory neural circuit leading to avoidance of hosts (smell and avoid hypothesis) or DEET has no behavioral effect on its own, but instead acts cooperatively with host odors to drive repellency (confusant hypothesis). Resolving this mystery will inform the search for a new generation of insect repellents.     

A theory of taste stimulation

The treatment in this paper of available quantitative data on the response of taste receptors to sodium salt stimulation clearly indicates that the ions of the chemical stimulus are loosely bound to some substance of the taste receptor. This can be thought of as an initial reaction which ultimately leads to stimulation of the receptor and an eventual depolarization of the associated sensory neuron. The speed of the total reaction suggests that the receptor substance is located on or near the surface of the receptor. The recently proposed (7) enzymatic reactions for chemoreceptors do not appear plausible for sodium salt stimulation of the taste receptors of the rat.     

Identification of olfactory receptor genes in Atlantic salmon Salmo salar

It has been hypothesized that salmonids use olfactory cues to return to their natal rivers and streams. The key components of the molecular pathways involved in imprinting and homing, however, are still unknown. Aquatic chemical cues are received through the nares and into the nasal cavity that contains a single olfactory organ, the olfactory rosette. The olfactory rosette contains sensory neurons, each of which is thought to express only one olfactory receptor. If odorants are involved in salmonid homing migration then olfactory receptors should play a critical role in the dissipation of information from the environment to the fish. Therefore, to understand the molecular basis for imprinting and homing in Atlantic salmon Salmo salar it is important to identify and characterize the repertoire of olfactory receptors in this species. The first public assembly of the S. salar genome was searched for genes encoding three of the superfamilies of fish olfactory receptors: V2R-like (olfc), V1R-like (ora) and main olfactory receptor (mor). A further six ora genes were added to ora1 and ora2, which had been described previously. In addition, 48 putative mors were identified, 24 of which appear to be functional based on their gene structures and predicted amino-acid sequences. Phylogenetic analyses were then used to compare these S. salar olfactory receptor genes with those of zebrafish Danio rerio, two pufferfish species Takifugu rubripes and Tetraodon nigroviridis, medaka Oryzias latipes and three-spined stickleback Gasterosteus aculeatus.     

Chemoreceptors in the cibario-pharyngeal pump of the cabbage looper moth,Trichoplusia ni (Lepidoptera: Noctuidae)

The structure of contact chemoreceptors in the cibariopharyngeal pump of the moth Trichoplusia ni (Lepidoptera: Noctuidae) is described. Two types of receptors designated A and B are located on the floor of the pump. Two groups of 9-12 A receptors are located in the anterior part of the pump, and two groups of two B receptors are in the posterior part of the pump. Five sensory dendrites extend to the tip of each A receptor and four to each B receptors. Available evidence indicates that these receptors are contact chemoreceptors and do not serve as mechanoreceptors. The receptors are compared to those of other insects.     

Intracellular trafficking of a tagged and functional mammalian olfactory receptor.

Tagged G-protein-coupled receptors (GPCRs) have been used to facilitate intracellular visualization of these receptors. We have used a combination of adenoviral vector gene transfer and tagged olfactory receptors to help visualize mammalian olfactory receptor proteins in the normal olfactory epithelium of rats, and in cell culture. Three recombinant adenoviral vectors were generated carrying variously tagged versions of rat olfactory receptor I7. The constructs include an N-terminal Flag epitope tag (Flag:I7), enhanced green fluorescent protein (EGFP) fusion protein (EGFP:I7), and a C-terminal EGFP fusion (I7:EGFP). These receptor constructs were assayed in rat olfactory sensory neurons (OSNs) and in a heterologous system (HEK 293 cell line) for protein localization and functional expression. Functional expression of the tagged receptor proteins was tested by electroolfactogram (EOG) recordings in the infected rat olfactory epithelium, and by calcium imaging in single cells. Our results demonstrate that the I7:EGFP fusion protein and Flag:I7 are functionally expressed in OSNs while the EGFP:I7 fusion is not, probably due to inappropriate processing of the protein in the cells. These data suggest that a small epitope tag (Flag) at the N-terminus, or EGFP located at the C-terminus of the receptor, does not affect ligand binding or downstream signaling. In addition, both functional fusion proteins (Flag:I7 and I7:EGFP) are properly targeted to the plasma membrane of HEK 293 cells.     

Olfactory receptors in Bactrocera species for sustainable fruit fly management: A review and future perspectives

Molecular studies on odorant receptors (ORs), odorant-binding proteins (OBPs) and the functioning of the receptor and pheromone signal transduction in fruit fly Bactrocera species have expanded exponentially during the past few decades. OBPs contribute to the sensing of the olfactory system (OS) via the transduction of odorants through the sensillum lymph. However, ORs, a family of G-protein-coupled receptors in Bactrocera and various other species, exhibit heightened responsiveness to multiple chemical odours such as hormones, sensory stimuli and neurotransmitters. The apparent mechanism involves a combinatorial code encompassing both peripheral and antennal lobe processing, facilitating the reception of sexual pheromones and environmental cues. The OS is specifically designed to recognize and process information from volatile chemical signals, and these chemical signals play an important function in various flies. Insects rely on these chemicals to navigate and comprehend their surroundings. A mature insect OS is composed of two pairs of sensillae-covered palps, antennae and two primary pairs of olfactory appendages on the anterior head. It has been shown that chemosensory gene families contribute in odour perception. These include various neuroreceptor families, such as OBPs, chemosensory proteins and sensory neuron membrane proteins. Additionally, there are three divergent chemoreceptors, namely ORs, ionotropic receptors and gustatory receptors. Methods based on systematic biology, molecular biology and bioinformatics tools have rapidly emerged to investigate the insect communication systems and provide new insights for the management of many agricultural pest. Several aromatic compounds, including semiochemicals and pheromones, have been employed to defend crops and animals from destructive fruit flies and other invasive and frugivorous species. To promote the expansion of the cropping system, the utilization of phytochemical lures can be convenient for sustainable agriculture production and enhance food security. Hence, this review examined the state of the art in chemical communication of insects with a focus on fruit fly pest species to identify OS and their semiochemical receptors, protein receptors and chemosensory receptors (CSRs), as well as their practical applications for biological control and integrated pest management are highlighted.     

Intracellular trafficking of a tagged and functional mammalian olfactory receptor

Tagged G‐protein‐coupled receptors (GPCRs) have been used to facilitate intracellular visualization of these receptors. We have used a combination of adenoviral vector gene transfer and tagged olfactory receptors to help visualize mammalian olfactory receptor proteins in the normal olfactory epithelium of rats, and in cell culture. Three recombinant adenoviral vectors were generated carrying variously tagged versions of rat olfactory receptor I7. The constructs include an N‐terminal Flag epitope tag (Flag:I7), enhanced green fluorescent protein (EGFP) fusion protein (EGFP:I7), and a C‐terminal EGFP fusion (I7:EGFP). These receptor constructs were assayed in rat olfactory sensory neurons (OSNs) and in a heterologous system (HEK 293 cell line) for protein localization and functional expression. Functional expression of the tagged receptor proteins was tested by electroolfactogram (EOG) recordings in the infected rat olfactory epithelium, and by calcium imaging in single cells. Our results demonstrate that the I7:EGFP fusion protein and Flag:I7 are functionally expressed in OSNs while the EGFP:I7 fusion is not, probably due to inappropriate processing of the protein in the cells. These data suggest that a small epitope tag (Flag) at the N‐terminus, or EGFP located at the C‐terminus of the receptor, does not affect ligand binding or downstream signaling. In addition, both functional fusion proteins (Flag:I7 and I7:EGFP) are properly targeted to the plasma membrane of HEK 293 cells. © 2002 Wiley Periodicals, Inc. J Neurobiol 50: 56–68, 2002     

Chemoreception and feeding behaviour in a caterpillar: towards a model of brain functioning in insects

The quantitative relationship between chemoreceptor activity during food intake and total amount of food ingested per unit of time has been studied in last instar larvae of Pieris brassicae. Data on sensory responses and feeding intensity, derived from a study by Blom (1978), show a strong correlation between receptor activity during 1 sec periods of stimulation and food intake during 24 h. It is concluded that nerve impulses from chemoreceptors, which signal the presence of different phagostimulants, are summated algebraically in the central nervous system. Impulses from feeding deterrent receptors counteract the effects of phagostimulants. One impulse from a deterrent receptor neutralizes 2.5 impulses from phagostimulant receptors. Based on the relationships observed a simple model of a feeding centre in the central nervous system has been constructed (Fig. 8).     

Lamellated outer dendritic segments of a chemoreceptor within wall-pore sensilla in the labial palp-pit organ of the butterfly,Pieris rapae L. (Insecta,Lepidoptera)

Summary The structure of the sensilla in the apical pit of the third segment of the labial palps in Pieris rapae was investigated in cryofixed and chemically fixed specimens. There is a field of about 80 club-shaped sensilla, 94% of which house a single sensory cell; 6% contain two sensory cells. All sensory cells are of the same type and are characterized by the structure of the dendritic outer segment. This consists of a proximal cylindrical and a distal lamellated section. The lamellae contain a lattice of longitudinally arranged microtubules. Filamentous strands connect the microtubules with the surface membrane of the lamellae. The surface area of the lamellated section amounts to about 40 m². Pores and pore tubules are present in the cuticular wall of the peg. Electrophysiological recordings show that the sensory cells are olfactory receptors, which react to a variety of complex plant odors and to the odor of conspecifics. It is shown that (a) the usual modality-specific characteristics of insect olfactory sensilla apply here also; (b) lamellation is not only a characteristic of thermoreceptors, but also of olfactory chemoreceptors; (c) there are pore tubules that are separated from the dendritic membranes by an extended dendritic sheath; and (d) in the labial palppit sensilla only the lamellated dendritic tip region may be involved in sensory transduction.Supported by the Deutsche Forschungsgemeinschaft (SFB 4/G1)