Philippe Litaudon

Optical recording of the rat piriform cortex activity

The piriform cortex (PCx) is a phylogenetically old brain structure which presents characteristics of a content-addressable memory. Taking into account its particular anatomo-functional organization, we hypothesized that this cortex could behave rather as an assembly of different functional units than as a functionally homogeneous structure. This hypothesis was tested by using both anatomical and functional approaches. Immunohistological and tracing experiments demonstrated that both the connections of the PCx with the higher nervous centres, and its monoaminergic and cholinergic modulatory afferents exhibited a heterogeneous distribution. Then, optical monitoring of its neuronal activity with a voltage-sensitive dye pointed out that the PCx is a functionally heterogeneous structure. Electrical stimulations of the olfactory bulb showed that the inhibitory processes which control the cortical responsiveness were not identical in all the PCx area. Two different functional areas at least could be distinguished: in the ventromedial PCx, the afferent activity is privileged since the level of inhibition of disynaptic activation remained large during repetitive stimuli. Contrarily, in the posterior PCx, the disynaptic activity remained unchanged in response to successive stimulations and the responses of neighbouring sites were statistically more synchronized than in its anterior part. Moreover, a late depolarization wave was significantly larger in the posterior PCx. These data are in good agreement with the results provided by computational models of the PCx. In the future, theoretical and experimental investigations of this cortex will be useful for understanding olfactory information processing and as a model of brain functioning at the neocortical level as well.

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.

Intrinsic association fiber system of the piriform cortex: A quantitative study based on a cholera toxin B subunit tracing in the rat

By using retrograde and anterograde transport of the B subunit of cholera toxin (CTb), we examined quantitatively the association fiber systems, i.e., the collaterals of pyramidal cell axons, that reciprocally connect both the rostral and the caudal parts of the piriform cortex (PC). Well‐defined CTb injections were obtained in layers Ib or II‐III of the rostral and the caudal parts of the PC. Using precision counting, we determined the proportion of cellular profiles in layers II and III that gave rise to association fibers and thus demonstrated a predominance of rostrocaudal fibers over the caudorostral ones. Our data also support a precise laminar organization of the PC in which the rostrocaudal fibers originated mainly from layer II and the caudorostral fibers primarily from layer III. Cholera toxin injections into layer Ib produced a peak of labeled profiles 2 mm from the site, indicating that a large proportion of the association fibers from layer II travel for at least 2 mm and then synapse in layer Ib. At either end of the PC, the association projections are concentrated laterally. The functional significance of these anatomical features is discussed with respect to olfactory processing, propagation of the activity within the PC, and the possible role of intrinsic fibers in olfactory memory.

fMRI visualization of transient activations in the rat olfactory bulb using short odor stimulations

Odor-evoked activity in the olfactory bulb displays both spatial and temporal organization. The difficulty when assessing spatio-temporal dynamics of olfactory representation is to find a method that reconciles the appropriate resolution for both dimensions. Imaging methods based on optical recordings can reach high temporal and spatial resolution but are limited to the observation of the accessible dorsal surface. Functional magnetic resonance imaging (fMRI) may be useful to overcome this limitation as it allows recording from the whole brain. In this study, we combined ultra fast imaging sequence and short stimulus duration to improve temporal resolution of odor-evoked BOLD responses. Short odor stimulations evoked high amplitude BOLD responses and patterns of activation were similar to those obtained in previous studies using longer stimulations. Moreover, short odor exposures prevented habituation processes. Analysis of the BOLD signal time course in the different areas of activation revealed that odorant response maps are not static entities but rather are temporally dynamic as reported by recent studies using optical imaging. These data demonstrated that fMRI is a non-invasive method which could represent a powerful tool to study not only the spatial dimension of odor representation but also the temporal dimension of information processing.

Olfactory Bulb Repetitive Stimulations Reveal Non-homogeneous Distribution of the Inhibitory Processes in the Rat Piriform Cortex

Optical signals were recorded in the in vivo rat piriform cortex in response to a burst of seven electrical stimulations (100 ms interval) delivered in the olfactory bulb. Based on the recorded responses, three types of signal could be identified according to the relative amplitude of their monosynaptic and disynaptic components. The disynaptic component had a larger (type 1) or an equal amplitude (type 2) compared with the monosynaptic one. Type 3 exhibited only the monosynaptic component. Type 1 represented 96% of the first response. The second response was characterized by an increase in type 3 signals (39%). The remaining type 1 signals were lower in amplitude when compared with the first response. The responses to the last five stimulations did not differ from one another but were different from the first two (type 1, 74%; type 2, 7.8%; type 3, 18.2% on average). The spatial distribution of these three types of signal was analysed by dividing the piriform cortex into several areas. These areas were not homogeneous in the percentage of each signal type: the percentage of type 3 signals was highest (˜30%) in the area near the lateral olfactory tract and <10% in the most posterodorsal area. Thus the level of inhibition remained high in some piriform areas whereas it decreased rapidly in others, suggesting that the inhibitory processes were not homogeneously distributed in the whole piriform cortex. Functional implications are discussed.

Piriform cortex late activity revealed functional spatial heterogeneity.

Optical signals were recorded in the in vivo rat piriform cortex (PC) in response to olfactory bulb electrical stimulation. Sometimes the early response was followed by a longer latency component with an occurrence probability of 0.25. In order to compare the early and late activity, the ratio between early (disynaptic) and late wave amplitudes was measured at each recording site on the whole PC. Its spatial distribution revealed that the relative importance of the late activity was larger in the most posterior part of the PC whereas the late wave was rarely observed in the anterior PC. Such a result gave new information on the functional heterogeneity of the PC.

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.

Role of the net architecture in piriform cortex activity: analysis by a mathematical model

Abstract. We present a mathematical analysis of the piriform cortex activity in rats. Experimental data were obtained by means of optical recording of fluorescent signals driven by neuronal activity. From these data, we determined the numerical value of the relaxation time for the pyramidal cell activity in layers II and III and the time latency map for bulb activation. Our model for the piriform cortex is based on pairs of excitatory and inhibitory neurons which correspond to pyramidal cells of layers II and III and to their inhibitory associated interneurons respectively; pyramidal cells are also interconnected through short and long range association fiber systems. Under such conditions, the model outputs resemble closely the experimental observations: (1) a double-bumped response to a strong and short stimulation; (2)␣oscillatory behavior under weak sustained stimulation conditions; (3) propagation of traveling activity waves; and (4) pacemaker activity when clusters of neurons are preferentially coupled.

Multi-site optical recording of the rat piriform cortex activity

Optical signals were recorded in the in vivo rat piriform cortex (PC) in response to olfactory bulb (OB) electrical stimulations delivered at 4 different sites. Afferent activity had a relatively wide (26.6% of the recorded area) but nonhomogeneous distribution on the PC surface. The different patterns of afferent activity observed in response to the 4 OB stimulations were intermixed with an overlap of only 38.5%. This activity was redistributed to the whole PC by intrinsic association fibres. The increase in the delay (from 4 ms to 12 ms) between afferent and redistributed activities along the antero-posterior axis indicated that the rostral to caudal association fibre system originating in the anterior PC was mainly responsible for the redistribution.