Robust odor coding via inhalation-coupled transient activity in the mammalian olfactory bulb

It has been proposed that a single sniff generates a "snapshot" of the olfactory world. However, odor coding on this timescale is poorly understood, and it is not known whether coding is invariant to changes in respiration frequency. We investigated this by recording spike trains from the olfactory bulb in awake, behaving rats. During rapid sniffing, odor inhalation triggered rapid and reliable cell- and odor-specific temporal spike patterns. These fine temporal responses conveyed substantial odor information within the first ～100 ms, and correlated with behavioral discrimination time on a trial-by-trial basis. Surprisingly, the initial transient portions of responses were highly conserved between rapid sniffing and slow breathing. Firing rates over the entire respiration cycle carried less odor information, did not correlate with behavior, and were poorly conserved across respiration frequency. These results suggest that inhalation-coupled transient activity forms a robust neural code that is invariant to changes in respiration behavior.     

Sniffing behavior of mice during performance in odor-guided tasks

Sniffing, a rhythmic inhalation and exhalation of air through the nose, is a behavior thought to play a critical role in shaping how odor information is represented and processed by the nervous system. Although the mouse has become a prominent model for studying olfaction, little is known about sniffing behavior in mice. Here, we characterized mouse sniffing behavior by measuring intranasal pressure transients in behaving mice. Sniffing was monitored during unstructured exploratory behavior and during performance of 3 commonly used olfactory paradigms: a habituation/dishabituation task, a sand digging-based discrimination task, and a nose poke-based discrimination task. We found that respiration frequencies in quiescent mice ranged from 3 to 5 Hz--higher than that reported for rats. During exploration, sniff frequency increased up to approximately 12 Hz and was highly dynamic, with rapid changes in frequency, amplitude, and waveform. Sniffing behavior varied strongly between tasks as well as for different behavioral epochs of each task. For example, mice performing the digging-based task showed little increase in sniff frequency prior to digging, whereas mice performing a nose poke-based task showed robust increases. Mice showed large increases in sniff frequency prior to reward delivery in all tasks. Mice also showed increases in sniff frequency when nose poking in a nonodor-guided task. These results show that mouse sniffing behavior is highly dynamic, varies with behavioral context, and is strongly modulated by olfactory as well as nonolfactory events.     

Anatomical contributions to odorant sampling and representation in rodents: zoning in on sniffing behavior

Odorant sampling behaviors such as sniffing bring odorant molecules into contact with olfactory receptor neurons (ORNs) to initiate the sensory mechanisms of olfaction. In rodents, inspiratory airflow through the nose is structured and laminar; consequently, the spatial distribution of adsorbed odorant molecules during inspiration is predictable. Physicochemical properties such as water solubility and volatility, collectively called sorptiveness, interact with behaviorally regulable variables such as inspiratory flow rate to determine the pattern of odorant deposition along the inspiratory path. Populations of ORNs expressing the same odorant receptor are distributed in strictly delimited regions along this inspiratory path, enabling different deposition patterns of the same odorant to evoke different patterns of neuronal activation across the olfactory epithelium and in the olfactory bulb. We propose that both odorant sorptive properties and the regulation of sniffing behavior may contribute to rodents' olfactory capacities by this mechanism. In particular, we suggest that the motor regulation of sniffing behavior is substantially utilized for purposes of "zonation" or the direction of odorant molecules to defined intranasal regions and hence toward distinct populations of receptor neurons, pursuant to animals' sensory goals.     

Alarm pheromones with different functions are released from different regions of the body surface of male rats

Our previous study suggested that the alarm pheromones in rats could be divided into at least two functionally different categories: one evoking autonomic responses and the other evoking behavioral responses, and the present study was conducted to test this hypothesis. Four regions of the body surface, i.e. the whisker pad, neck, rump and perianal region, of an anesthetized male Wistar rat were electrically stimulated (donor) and, after removal of the donor, the recipient rat was introduced into the same box and its behavioral and autonomic changes were recorded. Electrical stimulation of the perianal region of anesthetized donor rats provoked the release of odor that subsequently augmented core body temperature in other awake male rats. By contrast, electrical stimulation of the whisker pad of anesthetized donor males provoked the release of odor that augmented sniffing, rearing and locomotor activity in other awake male subjects. These results suggest that the alarm pheromone released from the face modifies behavior and that from the anal area induces autonomic stress responses in recipients.     

Multiday Recordings from Olfactory Bulb Neurons in Awake Freely Moving Rats: Spatially and Temporally Organized Variability in Odorant Response Properties

Chronic single-unit recordings were obtained from the mitral celllayer of the olfactory bulbs of awake freely moving rats placed in anodorant stream. Over periods up to five days, 618 recordings from 186single neurons were obtained. Responses of individual neurons werefound to be quite variable over time, although this variability wasbelow chance and was not incremental. The responses of nearbyneurons were more similar than expected by chance but less similarthan individual neurons recorded at different times. However,responses of spatially well-separated neurons were more differentthan chance over short time periods. During rapid sniffing,single-unit responses became more variable, and the spatialorganization of responses became less apparent. These results suggestthat neuronal responses in the olfactory bulb are generally quitevariable over time, with this variability increasing during periodsof rapid sniffing. These results are interpreted in the context of adistributed, centrally modulated model of olfactoryprocessing.     

Airflow behavior changes in upper airway caused by different head and neck positions: Comparison by computational fluid dynamics

The feasibility of computational fluid dynamics (CFD) to evaluate airflow characteristics in different head and neck positions has not been established. This study compared the changes in volume and airflow behavior of the upper airway by CFD simulation to predict the influence of anatomical and physiological airway changes due to different head–neck positions on mechanical ventilation. One awake volunteer with no risk of difficult airway underwent computed tomography in neutral position, extension position (both head and neck extended), and sniffing position (head extended and neck flexed). Three-dimensional airway models of the upper airway were reconstructed. The total volume (V) and narrowest area (A_min) of the airway models were measured. CFD simulation with an Spalart–Allmaras model was performed to characterize airflow behavior in neutral, extension, and sniffing positions of closed-mouth and open-mouth ventilation. The comparison result for V was neutral <extension≈sniffing, and for A_min was neutral<extension<sniffing. A_min in sniffing position was nearly 3.0 times that in neutral position and 1.7 times that in extension position. The pressure drop and velocity increasing were more obvious in neutral than sniffing or extension position at the same airflow rate. In sniffing position, pressure differences decreased and velocity remained almost constant. Recirculation airflow was generated near the subglottic region in neutral and extension positions. Sniffing position improves airway patency by increasing airway volume and decreasing airway resistance, suggesting that sniffing position may be the optimal choice for mask ventilation.     

Differences in the responses of identified neurons to chemical stimuli in satiated and hungry edible snails

Reactions of command neurones for avoidance behaviour to food were investigated in hungry and satiated snails in "CNS-chemoreceptor" preparations, in which hemolymph was washed out by saline. Neuronal responses in hungry animal preparations differed significantly from responses in satiated animals preparations. Perfusion of hungry animal preparations with hemolymph of satiated animals changed significantly the responses of command neurones for avoidance behaviour. These responses resembled the reactions of the same neurones to food after aversive conditioning to food of hungry snails. The role of humoral factor in learning is discussed.     

Distinct neuronal types contribute to hybrid temporal encoding strategies in primate auditory cortex

Studies of the encoding of sensory stimuli by the brain often consider recorded neurons as a pool of identical units. Here, we report divergence in stimulus-encoding properties between subpopulations of cortical neurons that are classified based on spike timing and waveform features. Neurons in auditory cortex of the awake marmoset (Callithrix jacchus) encode temporal information with either stimulus-synchronized or nonsynchronized responses. When we classified single-unit recordings using either a criteria-based or an unsupervised classification method into regular-spiking, fast-spiking, and bursting units, a subset of intrinsically bursting neurons formed the most highly synchronized group, with strong phase-locking to sinusoidal amplitude modulation (SAM) that extended well above 20 Hz. In contrast with other unit types, these bursting neurons fired primarily on the rising phase of SAM or the onset of unmodulated stimuli, and preferred rapid stimulus onset rates. Such differentiating behavior has been previously reported in bursting neuron models and may reflect specializations for detection of acoustic edges. These units responded to natural stimuli (vocalizations) with brief and precise spiking at particular time points that could be decoded with high temporal stringency. Regular-spiking units better reflected the shape of slow modulations and responded more selectively to vocalizations with overall firing rate increases. Population decoding using time-binned neural activity found that decoding behavior differed substantially between regular-spiking and bursting units. A relatively small pool of bursting units was sufficient to identify the stimulus with high accuracy in a manner that relied on the temporal pattern of responses. These unit type differences may contribute to parallel and complementary neural codes.

Neurons in auditory cortex show highly diverse responses to sounds. This study suggests that neuronal type inferred from baseline firing properties accounts for much of this diversity, with a subpopulation of bursting units being specialized for precise temporal encoding.     

Sensory-Motor Interactions for Vocal Pitch Monitoring in Non-Primary Human Auditory Cortex

The neural mechanisms underlying processing of auditory feedback during self-vocalization are poorly understood. One technique used to study the role of auditory feedback involves shifting the pitch of the feedback that a speaker receives, known as pitch-shifted feedback. We utilized a pitch shift self-vocalization and playback paradigm to investigate the underlying neural mechanisms of audio-vocal interaction. High-resolution electrocorticography (ECoG) signals were recorded directly from auditory cortex of 10 human subjects while they vocalized and received brief downward (−100 cents) pitch perturbations in their voice auditory feedback (speaking task). ECoG was also recorded when subjects passively listened to playback of their own pitch-shifted vocalizations. Feedback pitch perturbations elicited average evoked potential (AEP) and event-related band power (ERBP) responses, primarily in the high gamma (70–150 Hz) range, in focal areas of non-primary auditory cortex on superior temporal gyrus (STG). The AEPs and high gamma responses were both modulated by speaking compared with playback in a subset of STG contacts. From these contacts, a majority showed significant enhancement of high gamma power and AEP responses during speaking while the remaining contacts showed attenuated response amplitudes. The speaking-induced enhancement effect suggests that engaging the vocal motor system can modulate auditory cortical processing of self-produced sounds in such a way as to increase neural sensitivity for feedback pitch error detection. It is likely that mechanisms such as efference copies may be involved in this process, and modulation of AEP and high gamma responses imply that such modulatory effects may affect different cortical generators within distinctive functional networks that drive voice production and control.     

Simulated Birdwatchers’ Playback Affects the Behavior of Two Tropical Birds

Although recreational birdwatchers may benefit conservation by generating interest in birds, they may also have negative effects. One such potentially negative impact is the widespread use of recorded vocalizations, or “playback,” to attract birds of interest, including range-restricted and threatened species. Although playback has been widely used to test hypotheses about the evolution of behavior, no peer-reviewed study has examined the impacts of playback in a birdwatching context on avian behavior. We studied the effects of simulated birdwatchers’ playback on the vocal behavior of Plain-tailed Wrens Thryothorus euophrys and Rufous Antpittas Grallaria rufula in Ecuador. Study species’ vocal behavior was monitored for an hour after playing either a single bout of five minutes of song or a control treatment of background noise. We also studied the effects of daily five minute playback on five groups of wrens over 20 days. In single bout experiments, antpittas made more vocalizations of all types, except for trills, after playback compared to controls. Wrens sang more duets after playback, but did not produce more contact calls. In repeated playback experiments, wren responses were strong at first, but hardly detectable by day 12. During the study, one study group built a nest, apparently unperturbed, near a playback site. The playback-induced habituation and changes in vocal behavior we observed suggest that scientists should consider birdwatching activity when selecting research sites so that results are not biased by birdwatchers’ playback. Increased vocalizations after playback could be interpreted as a negative effect of playback if birds expend energy, become stressed, or divert time from other activities. In contrast, the habituation we documented suggests that frequent, regular birdwatchers’ playback may have minor effects on wren behavior.     

Classifying Response Correctness across Different Task Sets: A Machine Learning Approach

Erroneous behavior usually elicits a distinct pattern in neural waveforms. In particular, inspection of the concurrent recorded electroencephalograms (EEG) typically reveals a negative potential at fronto-central electrodes shortly following a response error (Ne or ERN) as well as an error-awareness-related positivity (Pe). Seemingly, the brain signal contains information about the occurrence of an error. Assuming a general error evaluation system, the question arises whether this information can be utilized in order to classify behavioral performance within or even across different cognitive tasks. In the present study, a machine learning approach was employed to investigate the outlined issue. Ne as well as Pe were extracted from the single-trial EEG signals of participants conducting a flanker and a mental rotation task and subjected to a machine learning classification scheme (via a support vector machine, SVM). Overall, individual performance in the flanker task was classified more accurately, with accuracy rates of above 85%. Most importantly, it was even feasible to classify responses across both tasks. In particular, an SVM trained on the flanker task could identify erroneous behavior with almost 70% accuracy in the EEG data recorded during the rotation task, and vice versa. Summed up, we replicate that the response-related EEG signal can be used to identify erroneous behavior within a particular task. Going beyond this, it was possible to classify response types across functionally different tasks. Therefore, the outlined methodological approach appears promising with respect to future applications.     

Maturation of sexual behavior in laboratory male mice: a role of genotype

Sexual behaviour and testosterone output in response to a receptive female were investigated in male mice of three inbred strains BALB/cLac, CBA/Lac and PT at puberty (45 days of age) and in adulthood (90 days of age). The animals were exposed for 10 min to a receptive female separated by a plastic grill, which would not allow contact between male and female. Male and female behaviour was recorded by measuring the time the male or female spent at the grill and the number of approaches to it (sexual motivation). The grill was then removed and the number of mounts and chemoinvestigatory behavior towards a female (nasal and anogenital sniffing) was recorded for each male. An increase in serum concentration and testicular content of testosterone was used as an endocrine index of the sensitivity to female pheromones. It has been shown the significant genotype and developmental effects on sexual behaviour and the hormonal response to sexual stimuli. The pubertal BALB/cLac males were characterised by the adult pattern of sexual motivation, chemoinvestigatory behaviour and the evident testosterone respond to a female. Males of the strain PT showed the lowest sexual motivation, chemoinvestigatory behavior towards a receptive female and no testosterone responses at both ages. This is a very different situation with the CBA/Lac's who showed the developmental increase in the sexual motivation, sniffing behaviour and the endocrine reflex, and the highest level of sexual behaviour but the moderate testosterone respond to a female at adulthood. The data obtained suggest genotype related asynchrony in maturation of the olfactory system, pituitary-gonadal axis and neural circuits of sexual behavior, and their independent genetic control. So, the set of mice strains investigated represents a useful tool for genetic and endocrine study of sexual behavior and the chemosensory control of testicular steroidogenesis.     

Variation in social relationships relates to song preferences and EGR1 expression in a female songbird

Social experiences can profoundly shape social behavior and the underlying neural circuits. Across species, the formation of enduring social relationships is associated with both neural and behavioral changes. However, it remains unclear how longer‐term relationships between individuals influence brain and behavior. Here, we investigated how variation in social relationships relates to variation in female preferences for and neural responses to song in a pair‐bonding songbird. We assessed variation in the interactions between individuals in male‐female zebra finch pairs and found that female preferences for their mate's song were correlated with the degree of affiliation and amount of socially modulated singing, but not with the frequency of aggressive interactions. Moreover, variation in measures of pair quality and preference correlated with variation in the song‐induced expression of EGR1, an immediate early gene related to neural activity and plasticity, in brain regions important for auditory processing and social behavior. For example, females with weaker preferences for their mate's song had greater EGR1 expression in the nucleus Taeniae, the avian homologue of the mammalian medial amygdala, in response to playback of their mate's courtship song. Our data indicate that the quality of social interactions within pairs relates to variation in song preferences and neural responses to ethologically relevant stimuli and lend insight into neural circuits sensitive to social information. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 76: 1029–1040, 2016     

Intranasal Odorant Concentrations in Relation to Sniff Behavior

Knowledge on how odorants are transported through the nasal cavity to the olfactory epithelium is limited. One facet of this is how the sniffing behavior affects the abundance of odorants transferred to the olfactory cleft and in turn influences odor perception. A novel system that couples an online mass spectrometer with an odorant pulse delivery olfactometer was employed to characterize intranasal odorant concentrations of butane‐2,3‐dione (or butanedione, commonly known as diacetyl) at the interior naris and the olfactory cleft. Volunteers (n=12) were asked to perform different modes of sniffing in relation to the sniff intensity that were categorized as ‘normal’, ‘rapid’ and ‘forced’. The highest concentrations of butanedione at both positions in the nose were observed during normal sniffing, with the lowest concentrations correlating with periods of forced sniffs. This corresponded to the panelists' ratings that normal sniffing elicited the highest odor intensities. These feasibility assessments pave the way for more in‐depth analyses with a variety of odorants of different chemical classes at various intranasal positions, to investigate the passage and uptake of odorants within the nasal cavity.     

Examination of Rapid Dopamine Dynamics with Fast Scan Cyclic Voltammetry During Intra-oral Tastant Administration in Awake Rats

Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique with high spatial and temporal (sub-second) resolution that has been utilized to examine phasic DA release in several types of preparations. In vitro experiments in single-cells and brain slices and in vivo experiments in anesthetized rodents have been used to identify mechanisms that mediate dopamine release and uptake under normal conditions and in disease models. Over the last 20 years, in vivo FSCV experiments in awake, freely moving rodents have also provided insight of dopaminergic mechanisms in reward processing and reward learning. One major advantage of the awake, freely moving preparation is the ability to examine rapid DA fluctuations that are time-locked to specific behavioral events or to reward or cue presentation. However, one limitation of combined behavior and voltammetry experiments is the difficulty of dissociating DA effects that are specific to primary rewarding or aversive stimuli from co-occurring DA fluctuations that mediate reward-directed or other motor behaviors. Here, we describe a combined method using in vivo FSCV and intra-oral infusion in an awake rat to directly investigate DA responses to oral tastants. In these experiments, oral tastants are infused directly to the palate of the rat – bypassing reward-directed behavior and voluntary drinking behavior – allowing for direct examination of DA responses to tastant stimuli.     

Is the hyperpnea of muscular contractions critically dependent on spinal afferents?

We studied the role of spinal afferent pathways in the hyperpnea of electrically induced muscle contractions (ExE). The ventilatory (VE) and arterial CO2 partial pressure (PaCO2) responses were measured at rest and during two levels of ExE in awake human paraplegic subjects with clinically complete lesions of the spinal cord (range T4-T11). We hypothesized that if peripheral neural drive is critical to a normal ventilatory response, then ExE in the absence of intact pathways should cause a lower ventilatory response resulting in hypercapnia at the onset of ExE. ExE was induced by stimulation of the quadriceps and hamstring muscles that approximately doubled the resting level of CO2 production (VCO2). PaCO2 during work transitions and in the latter stages of ExE did not differ significantly from that at rest. Arterial pH progressively declined over time during ExE (P less than 0.01) as a result of increased lactate concentration (P less than 0.01). The linear relationship between VE and VCO2 was similar to that found for normal human subjects during ExE (P = 0.73). These data suggest that VE and presumably alveolar ventilation (VA) can be appropriately matched to VCO2 during low-intensity muscle contractions of the lower extremities in the absence of intact spinal afferent pathways. Moreover, since it is unlikely that postulated "central command" mechanisms were initiated during ExE in these paraplegic subjects, the data provide support for our previous conclusion that central command is not obligatory for matching VA to VCO2 (J. Appl. Physiol. 64: 218-225, 1988).     

Inhibitory Control in Bilinguals and Musicians: Event Related Potential (ERP) Evidence for Experience-Specific Effects

Bilinguals and musicians exhibit behavioral advantages on tasks with high demands on executive functioning, particularly inhibitory control, but the brain mechanisms supporting these differences are unclear. Of key interest is whether these forms of experience influence cognition through similar or distinct information processing mechanisms. Here, we recorded event-related potentials (ERPs) in three groups – bilinguals, musicians, and controls – who completed a visual go-nogo task that involved the withholding of key presses to rare targets. Participants in each group achieved similar accuracy rates and responses times but the analysis of cortical responses revealed significant differences in ERP waveforms. Success in withholding a prepotent response was associated with enhanced stimulus-locked N2 and P3 wave amplitude relative to go trials. For nogo trials, there were altered timing-specific ERP differences and graded amplitude differences observed in the neural responses across groups. Specifically, musicians showed an enhanced early P2 response accompanied by reduced N2 amplitude whereas bilinguals showed increased N2 amplitude coupled with an increased late positivity wave relative to controls. These findings demonstrate that bilingualism and music training have differential effects on the brain networks supporting executive control over behavior.     

Cochlear Responses to Condensation and Rarefaction Clicks

Auditory nerve responses to condensation and rarefaction clicks (CC and RC) have been recorded over a wide intensity range with gross electrodes. At low intensities the RC responses are nearly identical to CC responses. At high intensities RC and CC response waveforms are similar, but the latency of the N₁ peak in the RC response is 0.2 msec. shorter than that for the corresponding CC response. At intermediate intensities the RC and CC response waveforms are quite different. These results can be interpreted in terms of a model in which there are two excitatory mechanisms for the neural response, which are operative in different intensity ranges. The cochlear microphonic potential and a “slow” potential are suggested as possible excitatory mechanisms.     

Diminished behavioral and neural sensitivity to sound modulation is associated with moderate developmental hearing loss

The acoustic rearing environment can alter central auditory coding properties, yet altered neural coding is seldom linked with specific deficits to adult perceptual skills. To test whether developmental hearing loss resulted in comparable changes to perception and sensory coding, we examined behavioral and neural detection thresholds for sinusoidally amplitude modulated (sAM) stimuli. Behavioral sAM detection thresholds for slow (5 Hz) modulations were significantly worse for animals reared with bilateral conductive hearing loss (CHL), as compared to controls. This difference could not be attributed to hearing thresholds, proficiency at the task, or proxies for attention. Detection thresholds across the groups did not differ for fast (100 Hz) modulations, a result paralleling that seen in humans. Neural responses to sAM stimuli were recorded in single auditory cortex neurons from separate groups of awake animals. Neurometric analyses indicated equivalent thresholds for the most sensitive neurons, but a significantly poorer detection threshold for slow modulations across the population of CHL neurons as compared to controls. The magnitude of the neural deficit matched that of the behavioral differences, suggesting that a reduction of sensory information can account for limitations to perceptual skills.     

A novel neural substrate for the transformation of olfactory inputs into motor output

It is widely recognized that animals respond to odors by generating or modulating specific motor behaviors. These reactions are important for daily activities, reproduction, and survival. In the sea lamprey, mating occurs after ovulated females are attracted to spawning sites by male sex pheromones. The ubiquity and reliability of olfactory-motor behavioral responses in vertebrates suggest tight coupling between the olfactory system and brain areas controlling movements. However, the circuitry and the underlying cellular neural mechanisms remain largely unknown. Using lamprey brain preparations, and electrophysiology, calcium imaging, and tract tracing experiments, we describe the neural substrate responsible for transforming an olfactory input into a locomotor output. We found that olfactory stimulation with naturally occurring odors and pheromones induced large excitatory responses in reticulospinal cells, the command neurons for locomotion. We have also identified the anatomy and physiology of this circuit. The olfactory input was relayed in the medial part of the olfactory bulb, in the posterior tuberculum, in the mesencephalic locomotor region, to finally reach reticulospinal cells in the hindbrain. Activation of this olfactory-motor pathway generated rhythmic ventral root discharges and swimming movements. Our study bridges the gap between behavior and cellular neural mechanisms in vertebrates, identifying a specific subsystem within the CNS, dedicated to producing motor responses to olfactory inputs.