Odor Fear Conditioning Modifies Piriform Cortex Local Field Potentials Both during Conditioning and during Post-Conditioning Sleep

BACKGROUND: Sleep plays an active role in memory consolidation. Sleep structure (REM/Slow wave activity [SWS]) can be modified after learning, and in some cortical circuits, sleep is associated with replay of the learned experience. While the majority of this work has focused on neocortical and hippocampal circuits, the olfactory system may offer unique advantages as a model system for exploring sleep and memory, given the short, non-thalamic pathway from nose to primary olfactory (piriform cortex), and rapid cortex-dependent odor learning. METHODOLOGY/PRINCIPAL FINDINGS: We examined piriform cortical odor responses using local field potentials (LFPs) from freely behaving Long-Evans hooded rats over the sleep-wake cycle, and the neuronal modifications that occurred within the piriform cortex both during and after odor-fear conditioning. We also recorded LFPs from naïve animals to characterize sleep activity in the piriform cortex and to analyze transient odor-evoked cortical responses during different sleep stages. Naïve rats in their home cages spent 40% of their time in SWS, during which the piriform cortex was significantly hypo-responsive to odor stimulation compared to awake and REM sleep states. Rats trained in the paired odor-shock conditioning paradigm developed enhanced conditioned odor evoked gamma frequency activity in the piriform cortex over the course of training compared to pseudo-conditioned rats. Furthermore, conditioned rats spent significantly more time in SWS immediately post-training both compared to pre-training days and compared to pseudo-conditioned rats. The increase in SWS immediately after training significantly correlated with the duration of odor-evoked freezing the following day. CONCLUSIONS/SIGNIFICANCE: The rat piriform cortex is hypo-responsive to odors during SWS which accounts for nearly 40% of each 24 hour period. The duration of slow-wave activity in the piriform cortex is enhanced immediately post-conditioning, and this increase is significantly correlated with subsequent memory performance. Together, these results suggest the piriform cortex may go offline during SWS to facilitate consolidation of learned odors with reduced external interference

The basolateral amygdala is necessary for the encoding and the expression of odor memory

Conditioned odor avoidance (COA) results from the association between a novel odor and a delayed visceral illness. The present experiments investigated the role of the basolateral amygdala (BLA) in acquisition and retrieval of COA memory. To address this, we used the GABA(A) agonist muscimol to temporarily inactivate the BLA during COA acquisition or expression. BLA inactivation before odor-malaise pairing greatly impaired COA tested 3 d later. In contrast, muscimol microinfusion between odor and malaise spared retention. Moreover, inactivation of the BLA before pre-exposure to the odor prevented latent inhibition of COA. This suggests that neural activity in the BLA is essential for the formation of odor representation. BLA inactivation before the retrieval test also blocked COA memory expression when performed either 3 d (recent memory) or 28 d (remote memory) after acquisition. This effect was transitory as muscimol-treated animals were not different from controls during the subsequent extinction tests. Moreover, muscimol infusion in the BLA neither affected olfactory perception nor avoidance behavior, and it did not induce a state-dependent learning. Altogether, these findings suggest that neural activity in the BLA is required for the encoding and the retrieval of odor memory. Moreover, the BLA seems to play a permanent role in the expression of COA

The basolateral amygdala is necessary for the encoding and the expression of odor memory

International audienceConditioned odor avoidance (COA) results from the association between a novel odor and a delayed visceral illness. The present experiments investigated the role of the basolateral amygdala (BLA) in acquisition and retrieval of COA memory. To address this, we used the GABA(A) agonist muscimol to temporarily inactivate the BLA during COA acquisition or expression. BLA inactivation before odor-malaise pairing greatly impaired COA tested 3 d later. In contrast, muscimol microinfusion between odor and malaise spared retention. Moreover, inactivation of the BLA before pre-exposure to the odor prevented latent inhibition of COA. This suggests that neural activity in the BLA is essential for the formation of odor representation. BLA inactivation before the retrieval test also blocked COA memory expression when performed either 3 d (recent memory) or 28 d (remote memory) after acquisition. This effect was transitory as muscimol-treated animals were not different from controls during the subsequent extinction tests. Moreover, muscimol infusion in the BLA neither affected olfactory perception nor avoidance behavior, and it did not induce a state-dependent learning. Altogether, these findings suggest that neural activity in the BLA is required for the encoding and the retrieval of odor memory. Moreover, the BLA seems to play a permanent role in the expression of COA

Long-term effects of infant learning on adult conditioned odor aversion are determined by the last preweaning experience

International audienceWe recently showed that odorizing mother's nipples from birth to weaning attenuated adult conditioned odor aversion (COA). The present study evaluated whether shorter durations of preweaning olfactory experiences could induce similar long-term effects. We first showed that late preweaning odorization (PN13-PN25) impaired adult COA similarly to odorization from birth to weaning (PN0-PN25) whereas early odorization (PN0-PN12) had no effect on adult COA. As early odorization was followed by an odorless suckling period, we evaluated whether this odorless suckling could have interfered with early associative learning. We therefore weaned the animals either immediately after early odorization or 7 days later. Early odorization (PN0-PN18) followed by late odorless suckling (PN19-PN25) had no effect on adult COA. However, pups with early odorization (PN0-PN18) but without late odorless suckling (weaned at PN18) showed attenuated COA. These results support the hypothesis that interference between early and late preweaning experiences with the mother determines the long-term impact on adult COA

Effets de l'odorisation des mamelles maternelles sur les apprentissages et comportements alimentaires olfactifs à l'âge adulte

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Critical role of insular cortex in taste but not odour aversion memory

International audienceConditioned odour aversion (COA) and conditioned taste aversion (CTA) result from the association of a novel odour or a novel taste with delayed visceral illness. The insular cortex (IC) is crucial for CTA memory, and the present experiments sought to determine whether the IC is required for the formation and the retrieval of COA memory as it is for CTA. We first demonstrated that ingested odour is as effective as taste for single-trial aversion learning in rats conditioned in their home cage. COA, like CTA, tolerates long intervals between the ingested stimuli and the illness and is long-lasting. Transient inactivation of the IC during acquisition spared COA whereas it greatly impaired CTA. Similarly, blockade of protein synthesis in IC did not affect COA but prevented CTA consolidation. Moreover, IC inactivation before retrieval tests did not interfere with COA memory expression when performed either 2 days (recent memory) or 36 days after acquisition (remote memory). Similar IC inactivation impaired the retrieval of either recent or remote CTA memory. Altogether these findings indicate that the IC is not necessary for aversive odour memory whereas it is essential for acquisition, consolidation and retrieval of aversive taste memory. We propose that the chemosensory stimulations modulate IC recruitment during the formation and the retrieval of food aversive memory

Rearing with artificially scented mothers attenuates conditioned odor aversion in adulthood but not its amygdala dependency

International audienceThe aim of the present study was to investigate whether neonatal odor experience associated with the mother affects food avoidance learning and basolateral amygdala (BLA) involvement in adulthood. Odorization of mother's nipples with banana or almond solutions from birth to weaning resulted in an impairment at adulthood of conditioned odor aversion (COA). These effects were specific to the early-experienced odor since no deficit was observed for COA to a novel odor (Experiment 1). In contrast, mere exposure to an odor in the home cage instead of on mother's nipples induced no deficit in COA at adulthood (Experiment 2). Finally, transitory inactivation of the BLA during COA acquisition in adult animals impaired the normal COA of naïve animals but also the attenuated COA of rats with early odor experience on the mother (Experiment 3). These results demonstrate that neonatal odor experience associated with the mother promotes the acquisition of appetitive memories which can interfere with food avoidance learning in adulthood. They also suggest that this early experience did not modify the BLA involvement in learned aversion

Involvement of CRFR1 in the basolateral amygdala in the immediate fear extinction deficit

Several animal and clinical studies have highlighted the ineffectiveness of fear extinction sessions delivered shortly after trauma exposure. This phenomenon, termed the immediate extinction deficit, refers to situations in which extinction programs applied shortly after fear conditioning may result in the reduction of fear behaviors (in rodents frequently measured as freezing responses to the conditioned cue/s) during extinction training, but failure to consolidate this reduction in the long-term. The molecular mechanisms driving this immediate extinction resistance remain unclear. Here we present evidence for the involvement of the corticotropin releasing factor (CRF) system in the basolateral amygdala (BLA) in male Wistar rats. Intra-BLA micro-infusion of the CRFR1 antagonist NBI30775 enhances extinction recall, while administering the CRF agonist CRF6-33 before delayed extinction disrupts recall of extinction. We link the immediate fear extinction deficit with dephosphorylation of GluA1 glutamate receptors at Ser845 and enhanced activity of the protein phosphatase calcineurin in the BLA. Their reversal following treatment with the CRFR1 antagonist indicates their dependency on CRFR1 actions. These findings can have important implications for the improvement of therapeutic approaches to trauma, as well as furthering our understanding of the neurobiological mechanisms underlying fear-related disorders. Significance Statement Trauma-related disorders are costly, highlighting the need to understand the reduction of fear through extinction learning for the development of better therapies. When extinction programs are applied too soon after the traumatic event, numerous studies have found it to be ineffective, though the underlying mechanisms were unclear. Here we confirm that futility of immediate extinction and provide a mechanistic explanation. Using a pharmacological approach, we show evidence for the involvement of the corticotropin releasing factor (CRF) system in the basolateral amygdala in this extinction deficit. We link this involvement with downstream molecular targets of the CRF system that are critical in synaptic plasticity, thus explaining the futility of immediate extinction and providing further insight into fear-related disorders