Rat Olfactory System Research Articles

Machine learning-based feature combination analysis for odor-dependent hemodynamic responses of rat olfactory bulb

Rodents have a well-developed sense of smell and are used to detect explosives, mines, illegal substances, hidden currency, and contraband, but it is impossible to keep their concentration constantly. Therefore, there is an ongoing effort to infer odors detected by animals without behavioral readings with brain-computer interface (BCI) technology. However, the invasive BCI technique has the disadvantage that long-term studies are limited by the immune response and electrode movement. On the other hand, near-infrared spectroscopy (NIRS)-based BCI technology is a non-invasive method that can measure neuronal activity without worrying about the immune response or electrode movement. This study confirmed that the NIRS-based BCI technology can be used as an odor detection and identification from the rat olfactory system. In addition, we tried to present features optimized for machine learning models by extracting six features, such as slopes, peak, variance, mean, kurtosis, and skewness, from the hemodynamic response, and analyzing the importance of individuals or combinations. As a result, the feature with the highest F1-Score was indicated as slopes, and it was investigated that the combination of the features including slopes and mean was the most important for odor inference. On the other hand, the inclusion of other features with a low correlation with slopes had a positive effect on the odor inference, but most of them resulted in insignificant or rather poor performance. The results presented in this paper are expected to serve as a basis for suggesting the development direction of the hemodynamic response-based bionic nose in the future.

Effects of lateral olfactory tract stimulation on Fos immunoreactivity in vasopressin neurones of the rat piriform cortex.

In the main olfactory system, odours are registered at the main olfactory epithelium and are then processed at the main olfactory bulb (MOB) and, subsequently, by the anterior olfactory nucleus (AON), the piriform cortex (PC) and the cortical amygdala. Previously, we reported populations of vasopressin neurones in different areas of the rat olfactory system, including the MOB, accessory olfactory bulb (AOB) and the AON and showed that these are involved in the coding of social odour information. Utilising immunohistochemistry and a transgenic rat in which an enhanced green fluorescent protein reporter gene is expressed in vasopressin neurones (eGFP-vasopressin), we now show a population of vasopressin neurones in the PC. The vasopressin neurones are predominantly located in the layer II of the PC and the majority co-express the excitatory transmitter glutamate. Furthermore, there is no sex difference in the number of neurones expressing vasopressin. Electrical stimulation of the lateral olfactory tract leads to a significant increase in the number of Fos-positive nuclei in the PC, MOB, AOB, dorsal AON and supraoptic nucleus (SON). However, there was only a significant increase in Fos expression in vasopressin cells of the PC and SON. Thus, functionally distinct populations of vasopressin cells are implicated in olfactory processing at multiple stages of the olfactory pathway.

Detection and classification of natural odors with an in vivo bioelectronic nose.

The mammalian olfactory system is recognized as one of the most effective chemosensing systems. We thus investigated the potential of utilizing the rat's olfactory system to detect odors. By chronically coupling multiple microelectrodes to olfactory bulb of behaving rats, we extract an array of mitral/tufted cells (M/Ts) which could generate odor-specific temporal patterns of neural discharge. We performed multidimensional analysis of recorded M/Ts, finding that natural odors released from different fruit lead to distinct odor response patterns. Thus an array of M/Ts carried sufficient information to discriminate odors. This novel brain-machine interface using rat's olfaction presents a promising method for odor detection and discrimination, and it is the first step towards in vivo bioelectronic nose equipped with biological olfaction and artificial devices.

The olfactory bulb theta rhythm follows all frequencies of diaphragmatic respiration in the freely behaving rat.

Sensory-motor relationships are part of the normal operation of sensory systems. Sensing occurs in the context of active sensor movement, which in turn influences sensory processing. We address such a process in the rat olfactory system. Through recordings of the diaphragm electromyogram (EMG), we monitored the motor output of the respiratory circuit involved in sniffing behavior, simultaneously with the local field potential (LFP) of the olfactory bulb (OB) in rats moving freely in a familiar environment, where they display a wide range of respiratory frequencies. We show that the OB LFP represents the sniff cycle with high reliability at every sniff frequency and can therefore be used to study the neural representation of motor drive in a sensory cortex.

Sex hormone binding globulin in the rat olfactory system

Ovarian steroids are known to act on the olfactory system. Their mode of action, however, is mostly unclear to date since nuclear receptors are lacking in sensory neurons. Here we used immunocytochemistry and RT-PCR to study expression and distribution of sex hormone binding globulin (SHBG) in the rat olfactory system. Single sensory cells in the olfactory mucosa and their projections in the olfactory bulb showed specific SHBG immunostaining as determined by double immunofluorescence with olfactory marker protein OMP. Larger groups of SHBG stained sensory cells occurred in the vomeronasal organ (VNO). A portion of the olfactory glomeruli in the accessory olfactory bulb showed large networks of SHBG positive nerve fibres. Some of the mitral cells showed SHBG immune fluorescence. RT-PCR revealed SHBG encoding mRNA in the olfactory mucosa, in the VNO and in the olfactory bulbs indicating intrinsic expression of the binding globulin. The VNO and its related projections within the limbic system are known to be sensitive to gonadal steroid hormones. We conclude that SHBG may be of functional importance for rapid effects of olfactory steroids on limbic functions including the control of reproductive behaviours through pheromones.

Laterality and Symmetry in Rat Olfactory Behavior and in Physiology of Olfactory Input

Many species use bilateral sampling for odor-guided navigation. Bilateral localization strategies typically involve balanced and lateralized sensory input and early neuronal processing. For example, if gradient direction is estimated by differential sampling, then any asymmetry could bias the perceived direction. Subsequent neuronal processing can compensate for this asymmetry but requires the presence of mechanisms to track changes in asymmetry. A high degree of laterality is also important for differential sampling because spillover of signals will dilute the perceived odor gradient. In apparent contradiction to this model, both symmetry and laterality of nasal air flow have been reported to be incomplete in rats. Here, we measured symmetry and laterality in early olfactory processing in the rat. We first established behavioral readouts of precisely controlled bilateral odorant stimuli. We found that rats could rapidly and accurately report the direction of a wide range of odor gradients, presented in random sequence. We then showed that nasal air flow was symmetric over an entire day in awake rats. Furthermore, odor sampling from the two nostrils in the behavioral task was highly lateralized. This lateralization extended to the receptor epithelium responses as measured by electro-olfactograms. We finally observed strong lateralization of intrinsic signal responses from the glomerular layer of the olfactory bulb. We confirmed that a differential comparison of glomerular responses was sufficient to localize odorants. Together, these results suggest that the rat olfactory system is symmetric, with highly lateralized odor flow and neuronal responses. In combination, these attributes support odor localization by differential comparison.

Acute toxicity of zinc oxide nanoparticles to the rat olfactory system after intranasal instillation

With the increased applications of zinc oxide (ZnO) nanoparticles (NPs), the toxicity of ZnO NPs arouses great concerns from the nano community and the general public. In this study, we report the toxicity of ZnO NPs (30 nm) to the rat olfactory system after intranasal instillation revealed by non-invasive magnetic resonance imaging (MRI). MRI scans were performed on a 4.7-T scanner at 1, 2, 3 and 7 days post-exposure, and the histological changes of the rat olfactory epithelium were evaluated. The influences of chemical component and dispersant of the NPs were also investigated. In addition, an olfactory behavior test was performed. The MRI and histological results indicated that ZnO NPs induced significant damages to the olfactory epithelium, including disruption of the olfactory epithelial structures and inflammation. The destruction of mitochondria in epithelial cells was observed under transmission electron microscopy (TEM), suggesting that the possible toxicological mechanism might involve cellular energy metabolic dysfunction. Further, the lesion of the olfactory epithelium disturbed sniffing behaviors of the treated animals. The results suggest that MRI is potentially useful as a screening tool to assess the consequence of occupational exposure of ZnO NPs. Caution should therefore be taken during the use and disposal of ZnO NPs to prevent the unintended public health impacts.

Expression of corticosteroid binding globulin in the rat olfactory system

Glucocorticoids are known to act on the olfactory system although their mode of action is still unclear since nuclear glucocorticoid receptors are mostly absent in the olfactory mucosa. In this study we used immunocytochemistry, in situ hybridization, and RT-PCR to study the expression and distribution of corticosteroid binding globulin (CBG) in the rat olfactory system. Mucosal goblet cells could be immunostained for CBG. Nasal secretion contained measurable amounts of CBG suggesting that CBG is liberated. CBG immunoreactivity was localized in many of the basal cells of the olfactory mucosa, while mature sensory cells contained CBG only in processes as determined by double immunostaining with the olfactory marker protein OMP. This staining was most pronounced in the vomeronasal organ (VNO). The appearance of CBG in the non-sensory and sensory parts of the VNO and in nerve terminals in the accessory bulb indicated axonal transport. Portions of the periglomerular cells, the mitral cells and the tufted cells were also CBG positive. CBG encoding transcripts were confirmed by RT-PCR in homogenates of the olfactory mucosa and VNO. Olfactory CBG may be significant for uptake, accumulation and transport of glucocorticoids, including aerosolic cortisol.

Localization of Nav1.7 in the normal and injured rodent olfactory system indicates a critical role in olfaction, pheromone sensing and immune function

Loss-of-function mutations in the pore-forming α subunit of the voltage-gated sodium channel 1.7 (Nav1.7) cause congenital indifference to pain and anosmia. We used immunohistochemical techniques to study Nav1.7 localization in the rat olfactory system in order to better understand its role in olfaction. We confirm that Nav1.7 is expressed on olfactory sensory axons and report its presence on vomeronasal axons, indicating an important role for Nav1.7 in transmission of pheromonal cues. Following neuroepithelial injury, Nav1.7 was transiently expressed by cells of monocytic lineage. These findings support an emerging role for Nav1.7 in immune function. This sodium channel may provide an important pharmacological target for treatment of inflammatory injury and inflammatory pain syndromes.

Temporally distinct expression of vesicular glutamate transporters 1 and 2 during embryonic development of the rat olfactory system

To study the development of glutamatergic neurons during the main olfactory bulb morphogenesis in rats, we examined the expression of vesicular glutamate transporters 1 (VGLUT1) and 2 (VGLUT2). On VGLUT1, expressions of mRNA and immunoreactivity were first detected in the mitral cell layer on embryonic day (E) 17.5 and E18.5, respectively, and persisted in the E20.5 olfactory bulb. Much earlier (on E12.5) than VGLUT1, expressions of VGLUT2 mRNA and/or immunoreactivity were found in the olfactory epithelium, migratory cells and telencephalon. On E14.5, the mRNA expression was also observed in the prospective bulbar region and vomeronasal organ, while immunoreactivity existed in migratory cells and growing fibers. Some fibers were observed in the deep telencephalic wall. From E16.5 onward, mRNA expression became gradually detectable in cells of the mitral cell layer with development. On E17.5, immunoreactivity was first found in fibers of the developing olfactory bulb and in some immature mitral cells from E18.5 to E20.5. The present study clarifies the expression of VGLUT2 precedent to VGLUT1 during olfactory bulb morphogenesis, suggesting differential contribution of the two VGLUT subtypes to glutamate-mediated embryonic events.

State-dependent functional connectivity of rat olfactory system assessed by fMRI

Functional connectivity between the piriform cortex and limbic and neocortical areas was assessed using functional magnetic resonance imaging (fMRI) of urethane anesthetized rats that spontaneously cycled between slow-wave and fast-wave states. Slow-wave and fast-wave states were determined indirectly through monitoring of respiration rate, which was confirmed to co-vary with state as determined by electrophysiological recordings. Previous electrophysiological data have suggested that the piriform cortex shifts between responsiveness to afferent odor input during fast-wave states and enhanced functional connectivity with limbic areas during slow-wave state. The present results demonstrate that fMRI-based resting state functional connectivity between the piriform cortex and both limbic and neocortical areas is enhanced during slow-wave state compared to fast-wave state using respiration as an indirect measure of state in urethane anesthetized rats. This state-dependent shift in functional connectivity may be important for sleep-dependent odor memory consolidation.

Associative Cortex Features in the First Olfactory Brain Relay Station

Synchronized firing of mitral cells (MCs) in the olfactory bulb (OB) has been hypothesized to help bind information together in olfactory cortex (OC). In this survey of synchronized firing by suspected MCs in awake, behaving vertebrates, we find the surprising result that synchronized firing conveys information on odor value ("Is it rewarded?") rather than odor identity ("What is the odor?"). We observed that asmice learned to discriminate between odors, synchronous firing responses to the rewarded and unrewarded odors became divergent. Furthermore, adrenergic blockage decreases the magnitude of odor divergence of synchronous trains, suggesting that MCs contribute to decision-making through adrenergic-modulated synchronized firing. Thus, in the olfactory system information on stimulus reward is found in MCs one synapse away from the sensory neuron.

Spike Timing Improves Olfactory Capabilities in Mammals

An issue that has puzzled neuroscientists for decades is what role, if any, temporal patterning of action potentials has in determining behavior. A study in this issue of Neuron by Cury and Uchida in the rat olfactory system provides evidence that such patterns could help mammals to identify and discriminate odors.

Sleep-Like States Modulate Functional Connectivity in the Rat Olfactory System

The present study was an examination of state-dependent functional connectivity during spontaneous activity between the piriform cortex and its upstream and downstream connections. Rats were anesthetized with urethan and allowed to spontaneously cycle between fast- and slow-wave states similar to fast- and slow-wave sleep states. Local field potential recordings were made from the olfactory bulb, piriform cortex, dorsal hippocampus, amygdala, and primary visual cortex. The results demonstrate that during slow-wave sleep-like states, when the piriform cortex shows reduced sensitivity to odor input via the olfactory bulb, there is enhanced coherence with other forebrain structures. Granger causality analyses suggest that the link between piriform cortical and hippocampal activity during slow-wave state is in the direction of the hippocampus to the piriform cortex rather than the reverse. The results suggest that slow-wave sleep-like states may provide an opportunity for the transfer and/or consolidation of information related to odor memories, specifically at a time when the piriform cortex is less sensitive to sensory input.

A three-compartment conductance-based model of the rat olfactory receptor neuron

Reduced compartmental models of single neurons, i.e. models with few Hodgkin-Huxley (HH) type compartments, provide an interesting balance between highly detailed, morphologically realistic models, and abstract, point-like models [1]. Simoes-de-Souza and Roque [2] constructed a four-compartment model of the vertebrate olfactory receptor neuron (ORN), which was used in a large-scale simulation of the olfactory system [3]. Their model contains a biochemical compartment, which allows the study of the role of molecular pathways in odor representation. However, the use of this compartment slows down computer simulations of models with many receptor cells and makes the analysis of network behavior more complicated – because of the large number of variables and parameters. In this work, we present a simpler ORN model which does not include biochemical pathways and contains only three HH type compartments. Our model is different from [2] not only because it has one compartment less but also because its passive and active parameters have new values adjusted by a combination of “hand-fitting” with automatic fitting procedures so as to reproduce well experimental results for the rat [5]. The model was constructed in GENESIS [4]. Its three compartments kept the same names used in [2]: soma, dendrite and dendritic knob. The latter two are passive compartments while soma has four voltage-gated ionic currents: high-voltage-activated calcium current, fast voltage- and calcium-dependent potassium current, slow calcium-dependent potassium current, and delayed rectifier potassium current. These ionic currents were modeled according to the standard HH formalism with kinetic parameters adjusted as mentioned above. Input stimuli were constant current steps injected directly in dendritic knob to simulate experimental results [5]. Some simulation results are shown in Figure ​Figure1.1. Our reduced three-compartment model may allow faster and more efficient large-scale network simulations of the rat olfactory system. Figure 1 Some simulation results. A. Spike train generated by a 10 pA step current (to be compared with Figure 2B of [5]); B. Action potential (to be compared with the inset of Figure 2A of [5]).

A Beta Oscillation Network in the Rat Olfactory System During a 2-Alternative Choice Odor Discrimination Task

We previously showed that in a two-alternative choice (2AC) task, olfactory bulb (OB) gamma oscillations (approximately 70 Hz in rats) were enhanced during discrimination of structurally similar odorants (fine discrimination) versus discrimination of dissimilar odorants (coarse discrimination). In other studies (mostly employing go/no-go tasks) in multiple labs, beta oscillations (15-35 Hz) dominate the local field potential (LFP) signal in olfactory areas during odor sampling. Here we analyzed the beta frequency band power and pairwise coherence in the 2AC task. We show that in a task dominated by gamma in the OB, beta oscillations are also present in three interconnected olfactory areas (OB and anterior and posterior pyriform cortex). Only the beta band showed consistently elevated coherence during odor sniffing across all odor pairs, classes (alcohols and ketones), and discrimination types (fine and coarse), with stronger effects in first than in final criterion sessions (>70% correct). In the first sessions for fine discrimination odor pairs, beta power for incorrect trials was the same as that for correct trials for the other odor in the pair. This pattern was not repeated in coarse discrimination, in which beta power was elevated for correct relative to incorrect trials. This difference between fine and coarse odor discriminations may relate to different behavioral strategies for learning to differentiate similar versus dissimilar odors. Phase analysis showed that the OB led both pyriform areas in the beta frequency band during odor sniffing. We conclude that the beta band may be the means by which information is transmitted from the OB to higher order areas, even though task specifics modify dominance of one frequency band over another within the OB.

Bestrophin 2: An anion channel associated with neurogenesis in chemosensory systems

The chemosensory neuroepithelia of the vertebrate olfactory system share a life-long ability to regenerate. Novel neurons proliferate from basal stem cells that continuously replace old or damaged sensory neurons. The sensory neurons of the mouse and rat olfactory system specifically express bestrophin 2, a member of the bestrophin family of calcium-activated chloride channels. This channel was recently proposed to operate as a transduction channel in olfactory sensory cilia. We raised a polyclonal antibody against bestrophin 2 and characterized the expression pattern of this protein in the mouse main olfactory epithelium, septal organ of Masera, and vomeronasal organ. Comparison with the maturation markers growth-associated protein 43 and olfactory marker protein revealed that bestrophin 2 was expressed in developing sensory neurons of all chemosensory neuroepithelia, but was restricted to proximal cilia in mature sensory neurons. Our results suggest that bestrophin 2 plays a critical role during differentiation and growth of axons and cilia. In mature olfactory receptor neurons, it appears to support growth and function of sensory cilia.

Spatio-temporal expression pattern of receptors for myelin-associated inhibitors in the developing rat olfactory system

The myelin-associated inhibitory proteins (Nogo-A, MAG and OMgp) that prevent axon regeneration in adult CNS, mediate their effects via a receptor referred as NgR1. Beside their inhibitory role in the adult CNS, Nogo-A and NgR1 might also be functionally involved in the developing nervous system. At the present time, no detailed study is available regarding either the onset of NgR1 expression during development or its spatio-temporal pattern of expression relative to the presence of Nogo-A. Two homologs of NgR1, NgR2 and NgR3, have been recently identified, but their function in the nervous system is still unknown in adult as well as during development. We have examined the spatio-temporal expression pattern of both NgR1, NgR2 and NgR3 mRNAs and corresponding proteins in the developing rat olfactory system using in situ hybridization and immunohistochemistry. From E15–E16 onwards, NgR1 mRNA was expressed by differentiating neurons in both the olfactory epithelium and the olfactory bulb. At all developmental stages, including adult animals, NgR1 protein was preferentially targeted to olfactory axons emerging from the olfactory epithelium. Using double-immunostainings in the postnatal olfactory mucosa, we confirm the neuronal localization of NgR1 and its preferential distribution along the olfactory axons. The NgR2 and NgR3 transcripts and their proteins display similar expression profiles in the olfactory system. Together, our data suggest that, in non-pathological conditions, NgR1 and its homologs may play a role in axon outgrowth in the rat olfactory system and may be relevant for the confinement of neural projections within the developing olfactory bulb.

Epithelial and endothelial barriers in the olfactory region of the nasal cavity of the rat

The olfactory ensheathing (glial) cells (OECs) have been identified to be useful candidate cells to support regeneration after being transplanted into injured fiber tracts of the central nervous system. We investigated by means of immunocytochemistry and freeze-fracturing the morphology and molecular composition of OEC tight junctions in the rat olfactory system. In addition, we tested the hypothesis whether tight junctions and orthogonal arrays of particles (OAPs) which contain the water channel protein aquaporin-4 (AQP4), are mutually exclusive as suggested in previous studies. In OECs, we found neither OAPs nor AQP4, but tight junctions immunoreactive for ZO-1, occludin, and claudin-5, but immunonegative for ZO-2 and claudin-3. To shed more light on the function of OEC tight junctions, we tested the permeability and tight junction composition of blood vessels and fila olfactoria. We found them both, permeable for infused lanthanum nitrate, and to be immunopositive for ZO-1 and claudin-5. The tight junctions of the OECs are discussed to be responsible for micro-compartmentalization within the olfactory fiber tract providing a benefit for axonal growth.

An Olfacto-Hippocampal Network Is Dynamically Involved in Odor-Discrimination Learning

Several studies have shown that memory consolidation relies partly on interactions between sensory and limbic areas. The functional loop formed by the olfactory system and the hippocampus represents an experimentally tractable model that can provide insight into this question. It had been shown previously that odor-learning associated beta band oscillations (15-30 Hz) of the local field potential in the rat olfactory system are enhanced with criterion performance, but it was unknown if these involve networks beyond the olfactory system. We recorded local field potentials from the olfactory bulb (OB) and dorsal and ventral hippocampus during acquisition of odor discriminations in a go/no-go task. These regions showed increased beta oscillation power during odor sampling, accompanied by a coherence increase in this frequency band between the OB and both hippocampal subfields. This coherence between the OB and the hippocampus increased with the onset of the first rule transfer to a new odor set and remained high for all learning phases and subsequent odor sets. However, coherence between the two hippocampal fields reset to baseline levels with each new odor set and increased again with criterion performance. These data support hippocampal involvement in the network underlying odor-discrimination learning and also suggest that cooperation between the dorsal and ventral hippocampus varies with learning progress. Oscillatory activity in the beta range may thus provide a mechanism by which these areas are linked during memory consolidation, similar to proposed roles of beta oscillations in other systems with long-range connections.