State-dependent alteration of respiration in a rat model of Parkinson's disease.

Parkinson's disease (PD) is the second most frequent neurodegenerative disorder. Besides major deficits in motor coordination, patients may also display sensory and cognitive impairments, which are often overlooked despite being inherently part of the PD symptomatology. Amongst those symptoms, respiration, a key mechanism involved in the regulation of multiple physiological and neuronal processes, appears to be altered. Importantly, breathing patterns are highly correlated with the animal's behavioral states. This raises the question of the potential impact of behavioral state on respiration deficits in PD. To answer this question, we first characterized the respiratory parameters in a neurotoxin-induced rat model of PD (6-OHDA) across three different vigilance states: sleep, quiet waking and exploration. We noted a significantly higher respiratory frequency in 6-OHDA rats during quiet waking compared to Sham rats. A higher respiratory amplitude was also observed in 6-OHDA rats during both quiet waking and exploration. No effect of the treatment was noted during sleep. Given the relation between respiration and olfaction and the presence of olfactory deficits in PD patients, we then investigated the odor-evoked sniffing response in PD rats, using an odor habituation/cross-habituation paradigm. No substantial differences were observed in olfactory abilities between the two groups, as assessed through sniffing frequency. These results corroborate the hypothesis that respiratory impairments in 6-OHDA rats are vigilance-dependent. Our results also shed light on the importance of considering the behavioral state as an impacting factor when analyzing respiration.

Distinct brain networks for remote episodic memory depending on content and emotional experience.

Memories of life episodes are the heart of individual stories. However, modelling episodic memory is a major challenge in both humans and animals when considering all its characteristics. As a consequence, the mechanisms that underlie the storage of old nontraumatic episodic memories remain enigmatic. Here, using a new task in rodents that models human episodic memory including odour/place/context components and applying advances behavioural and computational analyses, we show that rats form and recollect integrated remote episodic memories of two occasionally encountered complex episodes occurring in their daily life. Similar to humans, the information content and accuracy of memories vary across individuals and depend on the emotional relationship with odours experienced during the very first episode. We used cellular brain imaging and functional connectivity analyses, to find out the engrams of remote episodic memories for the first time. Activated brain networks completely reflect the nature and content of episodic memories, with a larger cortico-hippocampal network when the recollection is complete and with an emotional brain network related to odours that is critical in maintaining accurate and vivid memories. The engrams of remote episodic memories remain highly dynamic since synaptic plasticity processes occur during recall related to memory updates and reinforcement.

Respiration and brain neural dynamics associated with interval timing during odor fear learning in rats.

In fear conditioning, where a conditioned stimulus predicts the arrival of an aversive stimulus, the animal encodes the time interval between the two stimuli. Here we monitored respiration to visualize anticipatory behavioral responses in an odor fear conditioning in rats, while recording theta (5-15 Hz) and gamma (40-80 Hz) brain oscillatory activities in the medial prefrontal cortex (mPFC), basolateral amygdala (BLA), dorsomedial striatum (DMS) and olfactory piriform cortex (PIR). We investigated the temporal patterns of respiration frequency and of theta and gamma activity power during the odor-shock interval, comparing two interval durations. We found that akin to respiration patterns, theta temporal curves were modulated by the duration of the odor-shock interval in the four recording sites, and respected scalar property in mPFC and DMS. In contrast, gamma temporal curves were modulated by the interval duration only in the mPFC, and in a manner that did not respect scalar property. This suggests a preferential role for theta rhythm in interval timing. In addition, our data bring the novel idea that the respiratory rhythm might take part in the setting of theta activity dynamics related to timing.

High beta rhythm amplitude in olfactory learning signs a well-consolidated and non-flexible behavioral state.

Beta rhythm (15-30 Hz) is a major candidate underlying long-range communication in the brain. In olfactory tasks, beta activity is strongly modulated by learning but its condition of expression and the network(s) responsible for its generation are unclear. Here we analyzed the emergence of beta activity in local field potentials recorded from olfactory, sensorimotor and limbic structures of rats performing an olfactory task. Rats performed successively simple discrimination, rule transfer, memory recall tests and contingency reversal. Beta rhythm amplitude progressively increased over learning in most recorded areas. Beta amplitude reduced to baseline when new odors were introduced, but remained high during memory recall. Intra-session analysis showed that even expert rats required several trials to reach a good performance level, with beta rhythm amplitude increasing in parallel. Notably, at the beginning of the reversal task, beta amplitude remained high while performance was low and, in all tested animals, beta amplitude decreased before rats were able to learn the new contingencies. Connectivity analysis showed that beta activity was highly coherent between all structures where it was expressed. Overall, our results suggest that beta rhythm is expressed in a highly coherent network when context learning - including both odors and reward - is consolidated and signals behavioral inflexibility.

Adaptive quantization of local field potentials for wireless implants in freely moving animals: an open-source neural recording device.

OBJECTIVE: Modern neuroscience research requires electrophysiological recording of local field potentials (LFPs) in moving animals. Wireless transmission has the advantage of removing the wires between the animal and the recording equipment but is hampered by the large number of data to be sent at a relatively high rate. APPROACH: To reduce transmission bandwidth, we propose an encoder/decoder scheme based on adaptive non-uniform quantization. Our algorithm uses the current transmitted codeword to adapt the quantization intervals to changing statistics in LFP signals. It is thus backward adaptive and does not require the sending of side information. The computational complexity is low and similar at the encoder and decoder sides. These features allow for real-time signal recovery and facilitate hardware implementation with low-cost commercial microcontrollers. MAIN RESULTS: As proof-of-concept, we developed an open-source neural recording device called NeRD. The NeRD prototype digitally transmits eight channels encoded at 10 kHz with 2 bits per sample. It occupies a volume of 2 × 2 × 2 cm3 and weighs 8 g with a small battery allowing for 2 h 40 min of autonomy. The power dissipation is 59.4 mW for a communication range of 8 m and transmission losses below 0.1%. The small weight and low power consumption offer the possibility of mounting the entire device on the head of a rodent without resorting to a separate head-stage and battery backpack. The NeRD prototype is validated in recording LFPs in freely moving rats at 2 bits per sample while maintaining an acceptable signal-to-noise ratio (>30 dB) over a range of noisy channels. SIGNIFICANCE: Adaptive quantization in neural implants allows for lower transmission bandwidths while retaining high signal fidelity and preserving fundamental frequencies in LFPs.

Significance of sniffing pattern during the acquisition of an olfactory discrimination task.

Active sampling of olfactory environment consists of sniffing in rodents. The importance of sniffing dynamics is well established at the neuronal and behavioral levels. Patterns of sniffing have been shown to be modulated by the physicochemical properties of odorants, particularly concentration and sorption. Sniffing is also heavily impacted by higher processing related to the behavioral context, emotion and attentional demand. However, how the pattern of sniffing evolves over the course of learning of an experimental olfactory conditioning is still poorly understood. We tested this question by monitoring sniffing activity, using a whole-body plethysmograph, on rats performing a two-alternative choice odor discrimination task. We followed sniff variations at different learning stages (naïve, well-trained, expert). We found that during the acquisition of an odor discrimination task, rats acquired a global sniffing pattern, independent of the odor pair used. This pattern consists of a longer sampling duration, a higher sniffing frequency, and a larger amplitude. In parallel, subtle differences of sniffing between the two odors of a pair were also observed. This sniffing behavior was not only associated with a better and faster acquisition of the discrimination task but was also transferred to other odor sets and refined after a long-term pause so as to reduce the sampling duration and maintain a specific sniffing frequency. Our results provide additional arguments that sniffing is a complex sensorimotor act that is strongly affected by olfactory learning.

Memory of occasional events in rats: individual episodic memory profiles, flexibility, and neural substrate.

In search for the mechanisms underlying complex forms of human memory, such as episodic recollection, a primary challenge is to develop adequate animal models amenable to neurobiological investigation. Here, we proposed a novel framework and paradigm that provides means to quantitatively evaluate the ability of rats to form and recollect a combined knowledge of what happened, where it happened, and when or in which context it happened (referred to as episodic-like memory) after a few specific episodes in situations as close as possible to a paradigm we recently developed to study episodic memory in humans. In this task, rats have to remember two odor-drink associations (what happened) encountered in distinct locations (where it happened) within two different multisensory enriched environments (in which context/occasion it happened), each characterized by a particular combination of odors and places. By analyzing licking behavior on each drinking port, we characterized quantitatively individual recollection profiles and showed that rats are able to incidentally form and recollect an accurate, long-term integrated episodic-like memory that can last ≥ 24 d after limited exposure to the episodes. Placing rats in a contextually challenging recollection situation at recall reveals the ability for flexible use of episodic memory as described in humans. We further report that reversible inactivation of the dorsal hippocampus during recall disrupts the animal's capacity to recollect the complete episodic memory. Cellular imaging of c-Fos and Zif268 brain activation reveals that episodic memory recollection recruits a specific, distributed network of hippocampal-prefrontal cortex structures that correlates with the accuracy of the integrated recollection performance.

Olfactory preference conditioning changes the reward value of reinforced and non-reinforced odors.

Olfaction is determinant for the organization of rodent behavior. In a feeding context, rodents must quickly discriminate whether a nutrient can be ingested or whether it represents a potential danger to them. To understand the learning processes that support food choice, aversive olfactory learning and flavor appetitive learning have been extensively studied. In contrast, little is currently known about olfactory appetitive learning and its mechanisms. We designed a new paradigm to study conditioned olfactory preference in rats. After 8 days of exposure to a pair of odors (one paired with sucrose and the other with water), rats developed a strong and stable preference for the odor associated with the sucrose solution. A series of experiments were conducted to further analyze changes in reward value induced by this paradigm for both stimuli. As expected, the reward value of the reinforced odor changed positively. Interestingly, the reward value of the alternative odor decreased. This devaluation had an impact on further odor comparisons that the animal had to make. This result suggests that appetitive conditioning involving a comparison between two odors not only leads to a change in the reward value of the reinforced odor, but also induces a stable devaluation of the non-reinforced stimulus.

Sniff adjustment in an odor discrimination task in the rat: analytical or synthetic strategy?

A growing body of evidence suggests that sniffing is not only the mode of delivery for odorant molecules but also contributes to olfactory perception. However, the precise role of sniffing variations remains unknown. The zonation hypothesis suggests that animals use sniffing variations to optimize the deposition of odorant molecules on the most receptive areas of the olfactory epithelium (OE). Sniffing would thus depend on the physicochemical properties of odorants, particularly their sorption. Rojas-Líbano and Kay (2012) tested this hypothesis and showed that rats used different sniff strategies when they had to target a high-sorption (HS) molecule or a low-sorption (LS) molecule in a binary mixture. Which sniffing strategy is used by rats when they are confronted to discrimination between two similarly sorbent odorants remains unanswered. Particularly, is sniffing adjusted independently for each odorant according to its sorption properties (analytical processing), or is sniffing adjusted based on the pairing context (synthetic processing)? We tested these hypotheses on rats performing a two-alternative choice discrimination of odorants with similar sorption properties. We recorded sniffing in a non-invasive manner using whole-body plethysmography during the behavioral task. We found that sniffing variations were not only a matter of odorant sorption properties and that the same odorant was sniffed differently depending on the odor pair in which it was presented. These results suggest that rather than being adjusted analytically, sniffing is instead adjusted synthetically and depends on the pair of odorants presented during the discrimination task. Our results show that sniffing is a specific sensorimotor act that depends on complex synthetic processes.

A physiological increase of insulin in the olfactory bulb decreases detection of a learned aversive odor and abolishes food odor-induced sniffing behavior in rats.

Insulin is involved in multiple regulatory mechanisms, including body weight and food intake, and plays a critical role in metabolic disorders such as obesity and diabetes. An increasing body of evidence indicates that insulin is also involved in the modulation of olfactory function. The olfactory bulb (OB) contains the highest level of insulin and insulin receptors (IRs) in the brain. However, a role for insulin in odor detection and sniffing behavior remains to be elucidated. Using a behavioral paradigm based on conditioned olfactory aversion (COA) to isoamyl-acetate odor, we demonstrated that an intracerebroventricular (ICV) injection of 14 mU insulin acutely decreased olfactory detection of fasted rats to the level observed in satiated animals. In addition, whereas fasted animals demonstrated an increase in respiratory frequency upon food odor detection, this effect was absent in fasted animals receiving a 14 mU insulin ICV injection as well as in satiated animals. In parallel, we showed that the OB and plasma insulin levels were increased in satiated rats compared to fasted rats, and that a 14 mU insulin ICV injection elevated the OB insulin level of fasted rats to that of satiated rats. We further quantified insulin receptors (IRs) distribution and showed that IRs are preferentially expressed in the caudal and lateral parts of the main OB, with the highest labeling found in the mitral cells, the main OB projection neurons. Together, these data suggest that insulin acts on the OB network to modulate olfactory processing and demonstrate that olfactory function is under the control of signals involved in energy homeostasis regulation and feeding behaviors.

The relationship between respiration-related membrane potential slow oscillations and discharge patterns in mitral/tufted cells: what are the rules?

BACKGROUND: A slow respiration-related rhythm strongly shapes the activity of the olfactory bulb. This rhythm appears as a slow oscillation that is detectable in the membrane potential, the respiration-related spike discharge of the mitral/tufted cells and the bulbar local field potential. Here, we investigated the rules that govern the manifestation of membrane potential slow oscillations (MPSOs) and respiration-related discharge activities under various afferent input conditions and cellular excitability states. METHODOLOGY AND PRINCIPAL FINDINGS: We recorded the intracellular membrane potential signals in the mitral/tufted cells of freely breathing anesthetized rats. We first demonstrated the existence of multiple types of MPSOs, which were influenced by odor stimulation and discharge activity patterns. Complementary studies using changes in the intracellular excitability state and a computational model of the mitral cell demonstrated that slow oscillations in the mitral/tufted cell membrane potential were also modulated by the intracellular excitability state, whereas the respiration-related spike activity primarily reflected the afferent input. Based on our data regarding MPSOs and spike patterns, we found that cells exhibiting an unsynchronized discharge pattern never exhibited an MPSO. In contrast, cells with a respiration-synchronized discharge pattern always exhibited an MPSO. In addition, we demonstrated that the association between spike patterns and MPSO types appeared complex. CONCLUSION: We propose that both the intracellular excitability state and input strength underlie specific MPSOs, which, in turn, constrain the types of spike patterns exhibited.

The RUB Cage: Respiration-Ultrasonic Vocalizations-Behavior Acquisition Setup for Assessing Emotional Memory in Rats.

In animals, emotional memory is classically assessed through pavlovian fear conditioning in which a neutral novel stimulus (conditioned stimulus) is paired with an aversive unconditioned stimulus. After conditioning, the conditioned stimulus elicits a fear response characterized by a wide range of behavioral and physiological responses. Despite the existence of this large repertoire of responses, freezing behavior is often the sole parameter used for quantifying fear response, thus limiting emotional memory appraisal to this unique index. Interestingly, respiratory changes and ultrasonic vocalizations (USV) can occur during fear response, yet very few studies investigated the link between these different parameters and freezing. The aim of the present study was to design an experimental setup allowing the simultaneous recording of respiration, USV, and behavior (RUB cage), and the offline synchronization of the collected data for fine-grain second by second analysis. The setup consisted of a customized plethysmograph for respiration monitoring, equipped with a microphone capturing USV, and with four video cameras for behavior recording. In addition, the bottom of the plethysmograph was equipped with a shock-floor allowing foot-shock delivery, and the top received tubing for odor presentations. Using this experimental setup we first described the characteristics of respiration and USV in different behaviors and emotional states. Then we monitored these parameters during contextual fear conditioning and showed that they bring complementary information about the animal's anxiety state and the strength of aversive memory. The present setup may be valuable in providing a clearer appraisal of the physiological and behavioral changes that occur during acquisition as well as retrieval of emotional memory.

Reshaping of bulbar odor response by nasal flow rate in the rat.

BACKGROUND: The impact of respiratory dynamics on odor response has been poorly studied at the olfactory bulb level. However, it has been shown that sniffing in the behaving rodent is highly dynamic and varies both in frequency and flow rate. Bulbar odor response could vary with these sniffing parameter variations. Consequently, it is necessary to understand how nasal airflow can modify and shape odor response at the olfactory bulb level. METHODOLOGY AND PRINCIPAL FINDINGS: To assess this question, we used a double cannulation and simulated nasal airflow protocol on anesthetized rats to uncouple nasal airflow from animal respiration. Both mitral/tufted cell extracellular unit activity and local field potentials (LFPs) were recorded. We found that airflow changes in the normal range were sufficient to substantially reorganize the response of the olfactory bulb. In particular, cellular odor-evoked activities, LFP oscillations and spike phase-locking to LFPs were strongly modified by nasal flow rate. CONCLUSION: Our results indicate the importance of reconsidering the notion of odor coding as odor response at the bulbar level is ceaselessly modified by respiratory dynamics.

The way an odor is experienced during aversive conditioning determines the extent of the network recruited during retrieval: a multisite electrophysiological study in rats.

Recent findings have revealed the importance of orthonasal and retronasal olfaction in food memory, especially in conditioned odor aversion (COA); however, little is known about the dynamics of the cerebral circuit involved in the recognition of an odor as a toxic food signal and whether the activated network depends on the way (orthonasal vs retronasal) the odor was first experienced. In this study, we mapped the modulations of odor-induced oscillatory activities through COA learning using multisite recordings of local field potentials in behaving rats. During conditioning, orthonasal odor alone or associated with ingested odor was paired with immediate illness. For all animals, COA retrieval was assessed by orthonasal smelling only. Both types of conditioning induced similarly strong COA. Results pointed out (1) a predictive correlation between the emergence of powerful beta (15-40 Hz) activity and the behavioral expression of COA and (2) a differential network distribution of this beta activity according to the way the animals were exposed to the odor during conditioning. Indeed, for both types of conditioning, the aversive behavior was predicted by the emergence of a strong beta oscillatory activity in response to the odor in the olfactory bulb, piriform cortex, orbitofrontal cortex, and basolateral amygdala. This network was selectively extended to the infralimbic and insular cortices when the odor was ingested during acquisition. These differential networks could participate in different food odor memory; these results are discussed in line with recent behavioral results that indicate that COA can be formed over long odor-illness delays only if the odor is ingested.

A computer-assisted odorized hole-board for testing olfactory perception in mice.

The present paper describes a behavioral setup, designed and built in our laboratory, allowing the systematic and automatic recording of performances in a large number of olfactory behavioral tests. This computerized monitoring system has the capability of measuring different aspects of olfactory function in mice using different paradigms including threshold evaluation, generalization tasks, habituation/dishabituation, olfactory associative learning, short-term olfactory memory with or without a spatial component, and olfactory preferences. In this paper, we first describe the hole-board apparatus and its software and then give the experimental results obtained using this system. We demonstrate that one single, easy-to-run experimental setup is a powerful tool for the study of olfactory behavior in mice that has many advantages and broad applications.

Neonatal odor-shock conditioning alters the neural network involved in odor fear learning at adulthood.

Adult learning and memory functions are strongly dependent on neonatal experiences. We recently showed that neonatal odor-shock learning attenuates later life odor fear conditioning and amygdala activity. In the present work we investigated whether changes observed in adults can also be observed in other structures normally involved, namely olfactory cortical areas. For this, pups were trained daily from postnatal (PN) 8 to 12 in an odor-shock paradigm, and retrained at adulthood in the same task. (14)C 2-DG autoradiographic brain mapping was used to measure training-related activation in amygdala cortical nucleus (CoA), anterior (aPCx), and posterior (pPCx) piriform cortex. In addition, field potentials induced in the three sites in response to paired-pulse stimulation of the olfactory bulb were recorded in order to assess short-term inhibition and facilitation in these structures. Attenuated adult fear learning was accompanied by a deficit in 2-DG activation in CoA and pPCx. Moreover, electrophysiological recordings revealed that, in these sites, the level of inhibition was lower than in control animals. These data indicate that early life odor-shock learning produces changes throughout structures of the adult learning circuit that are independent, at least in part, from those involved in infant learning. Moreover, these enduring effects were influenced by the contingency of the infant experience since paired odor-shock produced greater disruption of adult learning and its supporting neural pathway than unpaired presentations. These results suggest that some enduring effects of early life experience are potentiated by contingency and extend beyond brain areas involved in infant learning.

Importance of retronasal and orthonasal olfaction for odor aversion memory in rats.

The role of odors in food memory formation, especially for aversions, has long been considered secondary to taste. However, the importance of odor ingestion in conditioned odor aversion (COA) has recently challenged this assumption (B. M. Slotnick, F. Westbrook, & F. M. C. Darling, 1997). The aim of the present study was to evaluate the respective role of orthonasal and retronasal olfactory experience in COA acquisition, long-term retention, extinction, and spontaneous recovery. To this end, the odor was presented either close to the drinking spout (orthonasal stimulation) or close to and mixed with the drinking water (eliciting both orthonasal and retronasal stimulation). The authors brought evidence that odor ingestion was crucial for COA acquisition, especially when odor presentation and gastric malaise were separated by long delays. On the contrary, once formed, a distal (orthonasal) odor recognition was sufficient for COA to be retrieved. COA was odor specific and long lasting (more than 50 days). Moreover, results brought evidence for a spontaneous recovery of odor aversion tested 57 days after its extinction.

Learning-induced oscillatory activities correlated to odour recognition: a network activity.

In trained behaving rats, the expression of a prominent beta oscillatory activity in the olfactory system was previously identified as a correlate of odour recognition. The aim of the present study was to assess the putative role of a functional coupling between the olfactory bulb (OB) and higher structures in this activity. We performed a unilateral inactivation of the medial part of the olfactory peduncle by lidocaine infusion. Inactivation deprived the OB from most of its centrifugal afferences, including feedback connections from the piriform cortex (PC) while sparing the ascending fibres from the OB to higher cortical structures. This treatment reduced the amplitude of odour-induced oscillatory beta responses both in OB and PC. In parallel, gamma activity classically observed in these two structures during spontaneous activity displayed a strong enhancement. Results suggest that odour-induced oscillatory response could be the emergent feature of an olfactory functional network set up through learning.

Olfactory fear conditioning induces field potential potentiation in rat olfactory cortex and amygdala.

The widely used Pavlovian fear-conditioning paradigms used for studying the neurobiology of learning and memory have mainly used auditory cues as conditioned stimuli (CS). The present work assessed the neural network involved in olfactory fear conditioning, using olfactory bulb stimulation-induced field potential signal (EFP) as a marker of plasticity in the olfactory pathway. Training consisted of a single training session including six pairings of an odor CS with a mild foot-shock unconditioned stimulus (US). Twenty-four hours later, the animals were tested for retention of the CS as assessed by the amount of freezing exhibited in the presence of the learned odor. Behavioral data showed that trained animals exhibited a significantly higher level of freezing in response to the CS than control animals. In the same animals, EFPs were recorded in parallel in the anterior piriform cortex (aPC), posterior piriform cortex (pPC), cortical nucleus of the amygdala (CoA), and basolateral nucleus of the amygdala (BLA) following electrical stimulation of the olfactory bulb. Specifically, EFPs recorded before (baseline) and after (during the retention test) training revealed that trained animals exhibited a lasting increase (present before and during presentation of the CS) in EFP amplitude in CoA, which is the first amygdaloid target of olfactory information. In addition, a transient increase was observed in pPC and BLA during presentation of the CS. These data indicate that the olfactory and auditory fear-conditioning neural networks have both similarities and differences, and suggest that the fear-related behaviors in each sensory system may have at least some distinct characteristics.

Learning modulation of odor-induced oscillatory responses in the rat olfactory bulb: a correlate of odor recognition?

In the first relay of information processing, the olfactory bulb (OB), odors are known to generate specific spatial patterns of activity. Recently, in freely behaving rats, we demonstrated that learning modulated oscillatory activity in local field potential (LFP), in response to odors, in both beta (15-40 Hz) and gamma (60-90 Hz) bands. The present study further characterized this odor-induced oscillatory activity with emphasis on its spatiotemporal distribution over the olfactory bulb and on its relationship with improvement of behavioral performances along training. For that purpose, LFPs were simultaneously recorded from four locations in the OB in freely moving rats performing an olfactory discrimination task. Electrodes were chronically implanted near relay neurons in the mitral cell body layer. Time-frequency methods were used to extract signal characteristics (amplitude, frequency, and time course) in the two frequency bands. Before training, odor presentation produced, on each site, a power decrease in gamma oscillations and a weak but significant increase in power of beta oscillations (approximately 25 Hz). When the training was achieved, these two phenomena were amplified. Interestingly, the beta oscillatory response showed several significant differences between the anterodorsal and posteroventral regions of the OB. In addition, clear-cut beta responses occurred in the signal as soon as animals began to master the task. As a whole, our results point to the possible functional importance of beta oscillatory activity in the mammalian OB, particularly in the context of olfactory learning.