PET Metabolic Imaging of Time-Dependent Reorganization of Olfactory Cued Fear Memory Networks in Rats.

Memory consolidation involves reorganization at both the synaptic and system levels. The latter involves gradual reorganization of the brain regions that support memory and has been mostly highlighted using hippocampal-dependent tasks. The standard memory consolidation model posits that the hippocampus becomes gradually less important over time in favor of neocortical regions. In contrast, this reorganization of circuits in amygdala-dependent tasks has been less investigated. Moreover, this question has been addressed using primarily lesion or cellular imaging approaches thus precluding the comparison of recent and remote memory networks in the same animals. To overcome this limitation, we used microPET imaging to characterize, in the same animals, the networks activated during the recall of a recent versus remote memory in an olfactory cued fear conditioning paradigm. The data highlighted the drastic difference between the extents of the two networks. Indeed, although the recall of a recent odor fear memory activates a large network of structures spanning from the prefrontal cortex to the cerebellum, significant activations during remote memory retrieval are limited to the piriform cortex. These results strongly support the view that amygdala-dependent memories also undergo system-level reorganization, and that sensory cortical areas might participate in the long-term storage of emotional memories.

Odorant features differentially modulate beta/gamma oscillatory patterns in anterior versus posterior piriform cortex.

Oscillatory activity is a prominent characteristic of the olfactory system. We previously demonstrated that beta and gamma oscillations occurrence in the olfactory bulb (OB) is modulated by the physical properties of the odorant. However, it remains unknown whether such odor-related modulation of oscillatory patterns is maintained in the piriform cortex (PC) and whether those patterns are similar between the anterior PC (aPC) and posterior PC (pPC). The present study was designed to analyze how different odorant molecular features can affect the local field potential (LFP) oscillatory signals in both the aPC and the pPC in anesthetized rats. As reported in the OB, three oscillatory patterns were observed: standard pattern (gamma + beta), gamma-only and beta-only patterns. These patterns occurred with significantly different probabilities in the two PC areas. We observed that odor identity has a strong influence on the probability of occurrence of LFP beta and gamma oscillatory activity in the aPC. Thus, some odor coding mechanisms observed in the OB are retained in the aPC. By contrast, probability of occurrence of different oscillatory patterns is homogeneous in the pPC with beta-only pattern being the most prevalent one for all the different odor families. Overall, our results confirmed the functional heterogeneity of the PC with its anterior part tightly coupled with the OB and mainly encoding odorant features whereas its posterior part activity is not correlated with odorant features but probably more involved in associative and multi-sensory encoding functions.

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.

Activity in the rat olfactory cortex is correlated with behavioral response to odor: a microPET study.

How olfactory cortical areas interpret odor maps evoked in the olfactory bulb and translate odor information into behavioral responses is still largely unknown. Indeed, rat olfactory cortices encompass an extensive network located in the ventral part of the brain, thus complicating the use of invasive functional methods. In vivo imaging techniques that were previously developed for brain activation studies in humans have been adapted for use in rodents and facilitate the non-invasive mapping of the whole brain. In this study, we report an initial series of experiments designed to demonstrate that microPET is a powerful tool to investigate the neural processes underlying odor-induced behavioral response in a large-scale olfactory neuronal network. After the intravenous injection of [18F]Fluorodeoxyglucose ([18F]FDG), awake rats were placed in a ventilated Plexiglas cage for 50 min, where odorants were delivered every 3 min for a 10-s duration in a random order. Individual behavioral responses to odor were classified into categories ranging from 1 (head movements associated with a short sniffing period in response to a few stimulations) to 4 (a strong reaction, including rearing, exploring and sustained sniffing activity, to several stimulations). After [18F]FDG uptake, rats were anesthetized to perform a PET scan. This experimental session was repeated 2 weeks later using the same animals without odor stimulation to assess the baseline level of activation in each individual. Two voxel-based statistical analyses (SPM 8) were performed: (1) a two-sample paired t test analysis contrasting baseline versus odor scan and (2) a correlation analysis between voxel FDG activity and behavioral score. As expected, the contrast analysis between baseline and odor session revealed activations in various olfactory cortical areas. Significant increases in glucose metabolism were also observed in other sensory cortical areas involved in whisker movement and in several modules of the cerebellum involved in motor and sensory function. Correlation analysis provided new insight into these results. [18F]FDG uptake was correlated with behavioral response in a large part of the anterior piriform cortex and in some lobules of the cerebellum, in agreement with the previous data showing that both piriform cortex and cerebellar activity in humans can be driven by sniffing activity, which was closely related to the high behavioral scores observed in our experiment. The present data demonstrate that microPET imaging offers an original perspective for rat behavioral neuroimaging.

Faster, deeper, better: the impact of sniffing modulation on bulbar olfactory processing.

A key feature of mammalian olfactory perception is that sensory input is intimately related to respiration. Different authors have considered respiratory dynamics not only as a simple vector for odor molecules but also as an integral part of olfactory perception. Thus, rats adapt their sniffing strategy, both in frequency and flow rate, when performing odor-related tasks. The question of how frequency and flow rate jointly impact the spatio-temporal representation of odor in the olfactory bulb (OB) has not yet been answered. In the present paper, we addressed this question using a simulated nasal airflow protocol on anesthetized rats combined with voltage-sensitive dye imaging (VSDi) of odor-evoked OB glomerular maps. Glomerular responses displayed a tonic component during odor stimulation with a superimposed phasic component phase-locked to the sampling pattern. We showed that a high sniffing frequency (10 Hz) retained the ability to shape OB activity and that the tonic and phasic components of the VSDi responses were dependent on flow rate and inspiration volume, respectively. Both sniffing parameters jointly affected OB responses to odor such that the reduced activity level induced by a frequency increase was compensated by an increased flow rate.

Individual and synergistic effects of sniffing frequency and flow rate on olfactory bulb activity.

Is faster or stronger sniffing important for the olfactory system? Odorant molecules are captured by sniffing. The features of sniffing constrain both the temporality and intensity of the input to the olfactory structures. In this context, it is clear that variations in both the sniff frequency and flow rate have a major impact on the activation of olfactory structures. However, the question of how frequency and flow rate individually or synergistically impact bulbar output has not been answered. We have addressed this question using multiple experimental approaches. In double-tracheotomized, anesthetized rats, we recorded both the bulbar local field potential (LFP) and mitral/tufted cells' activities when the sampling flow rate and frequency were controlled independently. We found that a tradeoff between the sampling frequency and the flow rate could maintain olfactory bulb sampling-related rhythmicity and that only an increase in flow rate could induce a faster, odor-evoked response. LFP and sniffing were recorded in awake rats. We found that sampling-related rhythmicity was maintained during high-frequency sniffing. Furthermore, we observed that the covariation between the frequency and flow rate, which was necessary for the tradeoff seen in the anesthetized preparations, also occurred in awake animals. Our study shows that the sampling frequency and flow rate can act either independently or synergistically on bulbar output to shape the neuronal message. The system likely takes advantage of this flexibility to adapt sniffing strategies to animal behavior. Our study provides additional support for the idea that sniffing and olfaction function in an integrated manner.

Respiration-gated formation of gamma and beta neural assemblies in the mammalian olfactory bulb.

A growing body of data suggests that information coding can be achieved not only by varying neuronal firing rate, but also by varying spike timing relative to network oscillations. In the olfactory bulb (OB) of a freely breathing anaesthetized mammal, odorant stimulation induces prominent oscillatory local field potential (LFP) activity in the beta (10-35 Hz) and gamma (40-80 Hz) ranges, which alternate during a respiratory cycle. At the same time, mitral/tufted (M/T) cells display respiration-modulated spiking patterns. Using simultaneous recordings of M/T unitary activities and LFP activity, we conducted an analysis of the temporal relationships between M/T cell spiking activity and both OB beta and gamma oscillations. We observed that M/T cells display a respiratory pattern that pre-tunes instantaneous frequencies to a gamma or beta regime. Consequently, M/T cell spikes become phase-locked to either gamma or beta LFP oscillations according to their frequency range and respiratory pattern. Our results suggest that slow respiratory dynamics pre-tune M/T cells to a preferential fast rhythm (beta or gamma) such that a spike-LFP coupling might occur when units and oscillation frequencies are in a compatible range. This double-coupling process might define two complementary beta- and gamma-neuronal assemblies along the course of a respiratory cycle.

Odor vapor pressure and quality modulate local field potential oscillatory patterns in the olfactory bulb of the anesthetized rat.

A central question in chemical senses is the way that odorant molecules are represented in the brain. To date, many studies, when taken together, suggest that structural features of the molecules are represented through a spatio-temporal pattern of activation in the olfactory bulb (OB), in both glomerular and mitral cell layers. Mitral/tufted cells interact with a large population of inhibitory interneurons resulting in a temporal patterning of bulbar local field potential (LFP) activity. We investigated the possibility that molecular features could determine the temporal pattern of LFP oscillatory activity in the OB. For this purpose, we recorded the LFPs in the OB of urethane-anesthetized, freely breathing rats in response to series of aliphatic odorants varying subtly in carbon-chain length or functional group. In concordance with our previous reports, we found that odors evoked oscillatory activity in the LFP signal in both the beta and gamma frequency bands. Analysis of LFP oscillations revealed that, although molecular features have almost no influence on the intrinsic characteristics of LFP oscillations, they influence the temporal patterning of bulbar oscillations. Alcohol family odors rarely evoke gamma oscillations, whereas ester family odors rather induce oscillatory patterns showing beta/gamma alternation. Moreover, for molecules with the same functional group, the probability of gamma occurrence is correlated to the vapor pressure of the odor. The significance of the relation between odorant features and oscillatory regimes along with their functional relevance are discussed.

A wavelet-based method for local phase extraction from a multi-frequency oscillatory signal.

One of the challenges in analyzing neuronal activity is to correlate discrete signal, such as action potentials with a signal having a continuous waveform such as oscillating local field potentials (LFPs). Studies in several systems have shown that some aspects of information coding involve characteristics that intertwine both signals. An action potential is a fast transitory phenomenon that occurs at high frequencies whereas a LFP is a low frequency phenomenon. The study of correlations between these signals requires a good estimation of both instantaneous phase and instantaneous frequency. To extract the instantaneous phase, common techniques rely on the Hilbert transform performed on a filtered signal, which discards temporal information. Therefore, time-frequency methods are best fitted for non-stationary signals, since they preserve both time and frequency information. We propose a new algorithmic procedure that uses wavelet transform and ridge extraction for signals that contain one or more oscillatory frequencies and whose oscillatory frequencies may shift as a function of time. This procedure provides estimates of phase, frequency and temporal features. It can be automated, produces manageable amounts of data and allows human supervision. Because of such advantages, this method is particularly suitable for analyzing synchronization between LFPs and unitary events.

Respiratory cycle as time basis: an improved method for averaging olfactory neural events.

In the mammalian olfactory system, neural activity appears largely modulated by respiration. Accurate analysis of respiratory synchronized activity is precluded by the variability of the respiratory frequency from trial to trial. Thus, the use of respiratory cycle as the time basis for measuring cell responses was developed about 20 years ago. Nevertheless, averaging oscillatory component of the activity remains a challenge due to their rhythmic features. In this paper, we present a new respiratory monitoring setup based on the measurement of micropressure changes induced by nasal airflow in front of the nostril. Improvements provided by this new monitoring setup allows automatic processing of respiratory signals in order to extract each respiratory period (expiration and inspiration). The time component of these periods, which can differ from trial to trial, is converted into a phase component defined as [-pi, 0] and [0, pi] for inspiration and expiration, respectively. As opposed to time representation, the phase representation is common to all trials. Thus, this phase representation of the respiratory cycle is used as a normalized time basis permitting to collect results in a standardized data format across different animals and providing new tools to average oscillatory components of the activity.

Respiratory modulation of olfactory neurons in the rodent brain.

In this review we report data from freely breathing animals in an attempt to show how respiratory dynamics can influence bulbar and cortical activity. Relying on in vivo data as well as in vitro observations, we try to emphasize the multiple mechanisms that underlie this modulation, its multiple origins, and its possible functional role.

Rhythm sequence through the olfactory bulb layers during the time window of a respiratory cycle.

The mammalian olfactory bulb is characterized by prominent oscillatory activity of its local field potentials. Breathing imposes the most important rhythm. Other rhythms have been described in the beta- and gamma-frequency ranges. We recorded unitary activities in different bulbar layers simultaneously with local field potentials in order to examine the different relationships existing between (i) breathing and field potential oscillations, and (ii) breathing and spiking activity of different cell types. We show that, whatever the layer, odour-induced gamma oscillations always occur around the transition point between inhalation and exhalation while beta oscillations appear during early exhalation and may extend up to the end of inhalation. By contrast, unitary activities exhibit different characteristics according to the layer. They vary in (i) their temporal relationship with respect to the respiratory cycle; (ii) their spike rates; (iii) their temporal patterns defined according to the respiratory cycle. The time window of a respiratory cycle might thus be split into three main epochs based on the deceleration of field potential rhythms (from gamma to beta oscillations) and a simultaneous gradient of spike discharge frequencies ranging from 180 to 30 Hz. We discuss the possibility that each rhythm could serve different functions as priming, gating or tuning for the bulbar network.