Primary Olfactory Neurons Research Articles

Dynamic Characterization ofDrosophilaAntennal Olfactory Neurons Indicates Multiple Opponent Signaling Pathways in Odor Discrimination

Opponent signaling refers to processes in which antagonistic signals are produced by different, but closely related, stimuli. It allows enhanced discrimination and more accurate behavioral responses. We explored opponent signaling in the Drosophila melanogaster olfactory system by measuring frequency response functions between odorant concentrations and primary olfactory neuron responses. Random fluctuations in concentration of two aliphatic and two aromatic fruit odorants were used to modulate action potentials from basiconic antennal sensilla. We separated action potentials by two-dimensional cluster analysis using amplitude and cross-correlation with a median action-potential template. Frequency response functions were fitted with either bandpass or second-order low-pass functions and then divided into two polarity groups, excitatory and inhibitory, by fitting the frequency response functions. Cluster analysis gave two, three, or four action potential clusters for each sensillum recording. Sensilla were then grouped by the patterns of response polarities of the individual neurons into four sensillum types, one with four neurons and three with two neurons. All four odorant compounds produced a mixture of excitatory, inhibitory, and null responses in different neurons. Statistical analysis of frequency response parameters for individual odorants gave only weak correlation between dynamics and some neuron types, even when comparing the dynamics of excitatory and inhibitory responses to the same odorant. However, response dynamics were significantly different between aliphatic and aromatic compounds, and between the two aliphatic compounds. Each odorant caused opposing excitatory and inhibitory signals to be sent to the antennal lobe along at least two pairs of axonal pathways.

Olfactory system and emotion: Common substrates

THE AIM OF THE REVIEW: A large number of studies suggest a close relationship between olfactory and affective information processing. Odors can modulate mood, cognition, and behavior. The aim of this article is to summarize the comparative anatomy of central olfactory pathways and centers involved in emotional analysis, in order to shed light on the relationship between the two systems. ANATOMY OF THE OLFACTORY SYSTEM: Odorant contact with the primary olfactory neurons is the starting point of olfactory transduction. The glomerulus of the olfactory bulb is the only relay between the peripheral and central olfactory system. Olfactory information is conducted to the secondary olfactory structures, notably the piriform cortex. The tertiary olfactory structures are the thalamus, hypothalamus, amygdala, hippocampus, orbitofrontal cortex and insular cortex. THE IMPACT OF ODORS ON AFFECTIVE STATES: Quality of life is commonly impaired in dysosmic patients. There have, however, been few publications on this topic. EMOTION AND OLFACTION: COMMON BRAIN PATHWAYS: There are brain structures common to emotion and odor processing. The present review focuses on such structures: amygdala, hippocampus, insula, anterior cingulate cortex and orbitofrontal cortex. The physiology and anatomy of each of these systems is described and discussed.

Microvillous cells expressing IP3 receptor type 3 in the olfactory epithelium of mice

Microvillous cells of the main olfactory epithelium have been described variously as primary olfactory neurons, secondary chemosensory cells or non-sensory cells. Here we generated an IP3R3(tm1(tauGFP)) mouse in which the coding region for a fusion protein of tau and green fluorescent protein replaces the first exon of the Itpr3 gene. We provide immunohistochemical and functional characterization of the cells expressing IP3 receptor type 3 in the olfactory epithelium. These cells bear microvilli at their apex, and we therefore termed them IP3R3 MV cells. The cell body of these IP3R3 MV cells lies in the upper third of the main olfactory epithelium; a long thick basal process projects towards the base of the epithelium without penetrating the basal lamina. Retrograde labeling and unilateral bulbectomy corroborated that these IP3R3 MV cells do not extend axons to the olfactory bulb and therefore are not olfactory sensory neurons. The immunohistochemical features of IP3R3 MV cells varied, suggesting either developmental stages or the existence of subsets of these cells. Thus, for example, subsets of the IP3R3 MV cells make contact with substance P fibers or express the purinergic receptor P2X3. In addition, in recordings of intracellular calcium, these cells respond to ATP and substance P as well as to a variety of odors. The characterization of IP3R3 MV cells as non-neuronal chemoresponsive cells helps to explain the differing descriptions of microvillous cells in the literature.

EphrinA5 protein distribution in the developing mouse brain

BackgroundEphrinA5 is one of the best-studied members of the Eph-ephrin family of guidance molecules, known to be involved in brain developmental processes. Using in situ hybridization, ephrinA5 mRNA expression has been detected in the retinotectal, the thalamocortical, and the olfactory systems; however, no study focused on the distribution of the protein. Considering that this membrane-anchored molecule may act far from the neuron soma expressing the transcript, it is of a crucial interest to localize ephrinA5 protein to better understand its function.ResultsUsing immunohistochemistry, we found that ephrinA5 protein is highly expressed in the developing mouse brain from E12.5 to E16.5. The olfactory bulb, the cortex, the striatum, the thalamus, and the colliculi showed high intensity of labelling, suggesting its implication in topographic mapping of olfactory, retinocollicular, thalamocortical, corticothalamic and mesostriatal systems. In the olfactory nerve, we found an early ephrinA5 protein expression at E12.5 suggesting its implication in the guidance of primary olfactory neurons into the olfactory bulb. In the thalamus, we detected a dynamic graduated protein expression, suggesting its role in the corticothalamic patterning, whereas ephrinA5 protein expression in the target region of mesencephalic dopaminergic neurones indicated its involvement in the mesostriatal topographic mapping. Following E16.5, the signal faded gradually and was barely detectable at P0, suggesting a main role for ephrinA5 in primary molecular events in topographic map formation.ConclusionOur work shows that ephrinA5 protein is expressed in restrictive regions of the developing mouse brain. This expression pattern points out the potential sites of action of this molecule in the olfactory, retinotectal, thalamocortical, corticothalamic and mesostriatal systems, during development. This study is essential to better understand the role of ephrinA5 during developmental topographic mapping of connections and to further characterise the mechanisms involved in pathway restoration following cell transplantation in the damaged brain.

Neuropeptide feedback modifies odor-evoked dynamics in Caenorhabditis elegans olfactory neurons.

Many neurons release classical transmitters together with neuropeptide cotransmitters whose functions are incompletely understood. Here we define the relationship between two transmitters in the olfactory system of Caenorhabditis elegans, showing that a neuropeptide-to-neuropeptide feedback loop alters sensory dynamics in primary olfactory neurons. The AWC olfactory neuron is glutamatergic and also expresses the peptide NLP-1. nlp-1 mutants have increased AWC-dependent behaviors, suggesting that NLP-1 limits the normal response. The receptor for NLP-1 is the G protein-coupled receptor NPR-11, which acts in postsynaptic AIA interneurons. Feedback from AIA interneurons modulates odor-evoked calcium dynamics in AWC olfactory neurons and requires INS-1, a neuropeptide released from AIA. The neuropeptide feedback loop dampens behavioral responses to odors on short and long timescales. Our results point to neuronal dynamics as a site of behavioral regulation and reveal the ability of neuropeptide feedback to remodel sensory networks on multiple timescales.

PHR1 is a vesicle‐bound protein abundantly expressed in mature olfactory neurons

To characterize the role of Phr1, a gene highly expressed in primary sensory neurons where it encodes an integral membrane protein with an N-terminal pleckstrin homology domain and a C-terminal transmembrane domain, in the olfactory system. We studied the immunelocalization of the PHR1 protein in mouse olfactory epithelium both at steady state and during regeneration following methyl bromide (MeBr) exposure using scanning confocal microscopy. Additionally, we examined the electrophysiologic role of Phr1 in olfaction and short-term olfactory adaptation. We found that PHR1 is abundantly and specifically expressed in olfactory neurons. It is widely distributed in punctate, vesiculated organelles throughout the cell bodies, axons, and glomeruli of primary olfactory neurons but is specifically excluded from the olfactory cilia. In the regenerating olfactory epithelium, PHR1 expression appears at 14 days following MeBr ablation coinciding with the onset of olfactory neuron maturity. Despite the abundant and specific expression throughout the olfactory neurons, mice lacking Phr1 did not exhibit differences in the distribution of the components of olfactory signal transduction system, the rate of olfactory regeneration following MeBr exposure, olfactory function, or short-term adaptation to odors. Phr1 is widely and abundantly expressed throughout mature olfactory neurons and other primary sensory neurons, but its absence does not appear to affect olfactory morphology, regeneration, sensory function, or adaptation. The exact function of Phr1 remains to be discovered.

The Alarm Reaction in Fishes—Odorants, Modulations of Responses, Neural Pathways

In 1938, von Frisch discovered the alarm reaction ("schreck reaktion") in fishes. He found that substances stored in the skin are released upon injury and detected by the olfactory organ. Several agonists that induce alarm reactions have been identified since then, but the true nature of the alarm substances remains veiled. Behavioral experiments have demonstrated that skin extract of conspecifics is more potent than that of heterospecifics. During development, the alarm reaction is seen in the larval stages in several cyprinid species; the threshold for eliciting the reaction decreases with age. Further, in crucian carp the alarm reaction was shown to be suppressed in the majority of females during the spawning period. The anatomy of the olfactory organ of this species is advantageous for studying underlying mechanisms of olfactory-induced behaviors. Crucian carp have been applied as a model organism in our lab for identification of the neural pathway of the alarm reaction, mainly in primary and secondary olfactory neurons. In the olfactory bulb, there is a distinct region that responds specifically to skin extract. The neural basis for discrimination between extracts of skin from different species has been investigated, suggesting that identification of conspecifics may be based on activation of specific subsets of neurons.

Role of IGF Signaling in Olfactory Sensory Map Formation and Axon Guidance

Olfactory neurons project their axons to spatially invariant glomeruli in the olfactory bulb, forming an ordered pattern of innervation comprising the olfactory sensory map. A mirror symmetry exists within this map, such that neurons expressing a given receptor typically project to one glomerulus on the medial face and one glomerulus on the lateral face of the bulb. The mechanisms underlying an olfactory neuron's choice to project medially versus laterally remain largely unknown, however. Here we demonstrate that insulin-like growth factor (IGF) signaling is required for sensory innervation of the lateral olfactory bulb. Mutations that eliminate IGF signaling cause axons destined for targets in the lateral bulb to shift to ectopic sites on the ventral-medial surface. Using primary cultures of olfactory and cerebellar neurons, we further show that IGF is a chemoattractant for axon growth cones. Together these observations reveal a role of IGF signaling in sensory map formation and axon guidance.

The shape of the olfactory bulb influences axon targeting

Each primary olfactory neuron in the mouse expresses a single type of odorant receptor. All neurons expressing the same odorant receptor gene typically project to two topographically fixed glomeruli, one each on the medial and lateral surfaces of the olfactory bulb. While topographic gradients of guidance receptors and their ligands help to establish the retinotectal projection, similar orthogonal distributions of cues have not yet been detected within the olfactory system. While odorant receptors are crucial for the final targeting of axons to glomeruli, it is unclear whether the olfactory bulb itself provides instructive cues for the establishment of the topographic map. To begin to understand the role of the olfactory bulb in the formation of the olfactory nerve pathway, we developed a model whereby the gross shape of the bulb in the P2-IRES-tau-LacZ line of mice was radically altered during postnatal development. We have shown here that the topography of axons expressing the P2 odorant receptor is dependent on the shape of the olfactory bulb. When the dorsoventral axis of the olfactory bulb was compressed during the early postnatal period, newly developing P2 axons projected to multiple inappropriate glomeruli surrounding their normal target site. These results suggest that the distribution of local guidance cues within the olfactory bulb is influenced by the shape of the olfactory bulb and that these cues contribute to the topographic positioning of glomeruli.

Predicting olfactory receptor neuron responses from odorant structure.

BackgroundOlfactory receptors work at the interface between the chemical world of volatile molecules and the perception of scent in the brain. Their main purpose is to translate chemical space into information that can be processed by neural circuits. Assuming that these receptors have evolved to cope with this task, the analysis of their coding strategy promises to yield valuable insight in how to encode chemical information in an efficient way.ResultsWe mimicked olfactory coding by modeling responses of primary olfactory neurons to small molecules using a large set of physicochemical molecular descriptors and artificial neural networks. We then tested these models by recording in vivo receptor neuron responses to a new set of odorants and successfully predicted the responses of five out of seven receptor neurons. Correlation coefficients ranged from 0.66 to 0.85, demonstrating the applicability of our approach for the analysis of olfactory receptor activation data. The molecular descriptors that are best-suited for response prediction vary for different receptor neurons, implying that each receptor neuron detects a different aspect of chemical space. Finally, we demonstrate that receptor responses themselves can be used as descriptors in a predictive model of neuron activation.ConclusionThe chemical meaning of molecular descriptors helps understand structure-response relationships for olfactory receptors and their "receptive fields". Moreover, it is possible to predict receptor neuron activation from chemical structure using machine-learning techniques, although this is still complicated by a lack of training data.

Intrinsically Bursting Olfactory Receptor Neurons

Rhythmically bursting neurons are fundamental to neuronal network function but typically are not considered in the context of primary sensory signaling. We now report intrinsically bursting lobster primary olfactory receptor neurons that respond to odors with a phase-dependent burst of action potentials. Rhythmic odor input as might be generated by sniffing entrains the intrinsic bursting rhythm in a concentration-dependent manner and presumably synchronizes the ensemble of bursting cells. We suggest such intrinsically bursting olfactory receptor cells provide a novel way for encoding odor information.

EARLY EVENTS IN OLFACTORY PROCESSING

Olfactory space has a higher dimensionality than does any other class of sensory stimuli, and the olfactory system receives input from an unusually large number of unique information channels. This suggests that aspects of olfactory processing may differ fundamentally from processing in other sensory modalities. This review summarizes current understanding of early events in olfactory processing. We focus on how odors are encoded by the activity of primary olfactory receptor neurons, how odor codes may be transformed in the olfactory bulb, and what relevance these codes may have for downstream neurons in higher brain centers. Recent findings in synaptic physiology, neural coding, and psychophysics are discussed, with reference to both vertebrate and insect model systems.

Olfactory experience accelerates glomerular refinement in the mammalian olfactory bulb

Primary olfactory sensory neurons (OSNs) impart both molecular and functional organization to the olfactory bulb. Because OSNs can be selectively activated by odorants in vivo, we sought to determine whether odorant experience alters the cellular dynamics of specific OSNs or their axonal projections. Using mice, we found that odorant stimulation associated with behavioral conditioning influenced OSN wiring by accelerating glomerular refinement independent of OSN number; furthermore, this wiring was strongly associated with olfactory learning.

The sorting behaviour of olfactory and vomeronasal axons during regeneration

In order to begin to understand how primary olfactory and vomeronasal organ (VNO) axons target specific regions of the olfactory bulb, we examined the sorting behaviour of these axons following neonatal unilateral olfactory bulbectomy. Bulbectomy induced widespread ipsilateral death of the primary olfactory and VNO neurons. After 4 weeks, many new sensory axons had re-grown into the cranial cavity and established a prominent plexus with evidence of dense tufts that were similar in gross appearance to glomeruli. Axons expressing the cell adhesion molecule OCAM, which normally innervate the ventrolateral and rostral halves of the main and accessory olfactory bulbs, respectively, sorted out and segregated from those axons not expressing this molecule within the plexus. In addition, VNO axons formed large discrete bundles that segregated from main olfactory axons within the plexus. Thus, VNO and primary olfactory axons as well as discrete subpopulations of both are able to sort out and remain segregated in the absence of the olfactory bulb. Sorting and convergence of axons therefore occur independently of the olfactory bulb and are probably attributable either to inherent properties of the axons themselves or to interactions between the axons and accompanying glial ensheathing cells.

Odorant receptors directly activate phospholipase C/inositol‐1,4,5‐trisphosphate coupled to calcium influx in Odora cells

Mechanisms by which odorants activate signaling pathways in addition to cAMP are hard to evaluate in heterogeneous mixtures of primary olfactory neurons. We used single cell calcium imaging to analyze the response to odorant through odorant receptor (OR) U131 in the olfactory epithelial cell line Odora (Murrell and Hunter 1999), a model system with endogenous olfactory signaling pathways. Because adenylyl cyclase levels are low, agents activating cAMP formation do not elevate calcium, thus unmasking independent signaling mediated by OR via phospholipase C (PLC), inositol-1,4,5-trisphosphate (IP(3)), and its receptor. Unexpectedly, we found that extracellular calcium is required for odor-induced calcium elevation without the release of intracellular calcium, even though the latter pathway is intact and can be stimulated by ATP. Relevant signaling components of the PLC pathway and G protein isoforms are identified by western blot in Odora cells as well as in olfactory sensory neurons (OSNs), where they are localized to the ciliary zone or cell bodies and axons of OSNs by immunohistochemistry. Biotinylation studies establish that IP(3) receptors type 2 and 3 are at the cell surface in Odora cells. Thus, individual ORs are capable of elevating calcium through pathways not directly mediated by cAMP and this may provide another avenue for odorant signaling in the olfactory system.

Brain and sensory organ morphology in Antarctic eelpouts (perciformes: Zoarcidae: Lycodinae)

Eelpouts of the family Zoarcidae comprise a monophyletic group of marine fishes with a worldwide distribution. Centers of high zoarcid diversity occur in the North Atlantic and North Pacific, with important radiations into the Arctic, along southern South America, and into the Southern Ocean around Antarctica. Along with snailfishes (Liparidae), zoarcids form an important component of the non-notothenioid fauna in the subzero shelf waters of Antarctica. We document the anatomy and histology of the brains, cranial nerves, olfactory apparatus, cephalic lateral lines, taste buds, and retinas of three Antarctic zoarcid species, living at depths of 310-939 m, representing three of the nine genera from this region. The primary emphasis is on Ophthalmolycus amberensis, and we provide a detailed drawing of the brain and cranial nerves of this species. Although this brain reflects general perciform neural morphology, it exhibits a reduction of the (optic) tecta and the eminentia granulares and crista cerebellares of the lateral line system. Interspecific differences among the three species are slight. The olfactory rosette consists of three to four lamellae and the nasal sac, contrary to the claim of Fanta et al. ([2001] Antarct Rec, Natl Inst Polar Res, Tokyo 45:27-42), is not in communication with the cephalic lateral line system. Primary olfactory neurons are abundant and converge on branches of the olfactory nerve. Numerous taste buds are located in the lips. All three species lack an ocular choroid rete and have relatively thin retinas with a low cell density and a single bank of rods as the only type of photoreceptor. Neural diversification among Antarctic zoarcids has not involved the evolution of sensory specialists; brain and sensory organ morphologies do not approach the condition seen in primary deep-sea fishes, or even that of some sympatric non-perciform secondary deep-sea fishes, including liparids and muraenolepidids (eel cods). There may be phylogenetic constraints on brain morphology in perciforms such that we do not see extreme specialization in sensory and neural systems for deep habitats. We suggest that the brains and sensory organs of Antarctic zoarcids reflect habitation of 500-2,000-m depths and likely reflect morphologies seen in zoarcids living on continental slopes elsewhere in the world. This balance among the sensory modalities makes zoarcids relatively generalized among secondary deep-sea fishes and may be one of the reasons this opportunistic and adaptable group has been successful in colonizing a variety of emergent and ephemeral habitats.

Isolation and characterization of the laure olfactory behavioral mutant in the zebrafish, Danio rerio

To initiate a genetic analysis of olfactory development and function in the zebrafish, Danio rerio, we developed a behavioral genetic screen for mutations affecting the olfactory sensory system. First, we characterized olfactory responses of wild-type zebrafish to various odors. We found that 3-day-old juvenile zebrafish reacted to the amino acid L-cysteine with an aversive behavioral response. We isolated one mutant, laure (lre), which showed no aversive behavioral response to L-cysteine at 3 days of development, and carried out a preliminary characterization of this mutant's defects. We found that lre mutant fish were also defective in their response to L-serine and L-alanine, but not to taurocholic acid, as young adults. In addition, lre mutant fish had significantly fewer primary olfactory sensory neurons than normal, and the axons of these neurons did not form the characteristic axon termination pattern in the developing olfactory bulb. Nevertheless, the olfactory epithelium of lre mutant fish showed normal or near normal electrophysiological responses to several odorants. Our data suggest that the behavioral defects observed in the lre mutant result from the disruption of the developing olfactory sensory neurons and their axonal connections within the olfactory bulb. The isolation of the lre mutant shows that our behavior-based screen represents a viable approach for carrying out a genetic dissection of olfactory behaviors in this vertebrate model system.

Olfactory marker protein modulates primary olfactory axon overshooting in the olfactory bulb

Olfactory marker protein (OMP) is expressed by mature primary olfactory sensory neurons during development and in adult mice. In mice that lack OMP, olfactory sensory neurons have perturbed electrophysiological activity, and the mice exhibit altered responses and behavior to odor stimulation. To date, defects in axon guidance in mice that lack OMP have not been investigated. During development of the olfactory system in mouse, primary olfactory axons often overshoot their target glomerular layer and project into the deeper external plexiform layer. These aberrant axonal projections are normally detected within the external plexiform layer up to postnatal day 12. We have examined the projections of primary olfactory axons in OMP-tau:LacZ mice and OMP-GFP mice, two independent lines in which the OMP coding region has been replaced by reporter molecules. We found that axons overshoot their target layer and grow into the external plexiform layer in these OMP null mice as they do in wild-type animals. However, in the absence of OMP, overshooting axons are more persistent and remain prominent until 5 weeks postnatally, after which their numbers decrease. Overshooting axons are still present in these mice even at 8 months of age. In heterozygous mice, axons also overshoot into the external plexiform layer; however, there are fewer axons, and they project for shorter distances, compared with those in a homozygous environment. Our results suggest that perturbed electrophysiological responses, caused by loss of OMP in primary olfactory neurons, reduce the ability of primary olfactory axons to recognize their glomerular target.

Immunolocalisation of P2X and P2Y nucleotide receptors in the rat nasal mucosa

Purinoceptor subtypes were localised to various tissue types present within the nasal cavity of the rat, using immunohistochemical methods. P2X3 receptor immunoreactivity was localised in the primary olfactory neurones located both in the olfactory epithelium and vomeronasal organs (VNO) and also on subepithelial nerve fibres in the respiratory region. P2X5 receptor immunoreactivity was found in the squamous, respiratory and olfactory epithelial cells of the rat nasal mucosa. P2X7 receptor immunoreactivity was also expressed in epithelial cells and colocalised with caspase 9 (an apoptotic marker), suggesting an association with apoptosis and epithelial turnover. P2Y1 receptor immunoreactivity was found within the respiratory epithelium and submucosal glandular tissue. P2Y2 receptor immunoreactivity was localised to the mucus-secreting cells within the VNO. The possible functional roles of these receptors are discussed.

Extrabulbar olfactory system and nervus terminalis FMRFamide immunoreactive components in Xenopus laevis ontogenesis

The extrabulbar olfactory system (EBOS) is a collection of nerve fibers which originate from primary olfactory receptor-like neurons and penetrate into the brain bypassing the olfactory bulbs. Our description is based upon the application of two neuronal tracers (biocytin, carbocyanine DiI) in the olfactory sac, at the cut end of the olfactory nerve and in the telencephalon of the developing clawed frog. The extrabulbar olfactory system was observed already at stage 45, which is the first developmental stage compatible with our techniques; at this stage, the extrabulbar olfactory system fibers terminated diffusely in the preoptic area. A little later in development, i.e. at stage 50, the extrabulbar olfactory system was maximally developed, extending as far caudally as the rhombencephalon. In the metamorphosing specimens, the extrabulbar olfactory system appeared reduced in extension; caudally, the fiber terminals did not extend beyond the diencephalon. While a substantial overlapping of biocytin/FMRFamide immunoreactivity was observed along the olfactory pathways as well as in the telencephalon, FMRFamide immunoreactivity was never observed to be colocalized in the same cellular or fiber components visualized by tracer molecules. The question whether the extrabulbar olfactory system and the nervus terminalis (NT) are separate anatomical entities or represent an integrated system is discussed.