La plasticité synaptique est une propriété indispensable à l’acquisition de la mémoire chez toutes les espèces étudiées, des invertébrés aux primates. La formation d’une mémoire débute par une phase de plasticité qui inclut une restructuration synaptique ; ensuite elle se poursuit par la consolidation de ces modifications, contribuant à la mémoire à long terme. Certaines mémoires redeviennent malléables lorsqu’elles sont rappelées. La trace mnésique entre alors dans une nouvelle de phase de plasticité, au cours de laquelle certaines composantes de la mémoire peuvent être mises à jour, puis reconsolidées. L’objectif de la présente thèse est d’étudier les mécanismes cellulaires et moléculaires qui sont activés lors du rappel d’une mémoire. Nous avons utilisé un modèle de conditionnement Pavlovien, combiné à l’administration d’agents pharmacologiques et à l’analyse quantitative de marqueurs de plasticité synaptique, afin d’étudier la dynamique de la mémoire de peur auditive chez des rats Sprague Dawley. La circuiterie neuronale et les mécanismes associatifs impliqués dans la neurobiologie de cette mémoire sont bien caractérisés, en particulier le rôle des récepteurs glutamatergiques de type NMDA et AMPA dans la plasticité synaptique et la consolidation. Nos résultats démontrent que le retour de la trace mnésique à un état de labilité nécessite l’activation des récepteurs NMDA dans l’amygdale baso-latérale à l’instant même du rappel, alors que les récepteurs AMPA sont requis pour l’expression comportementale de la réponse de peur conditionnée. D’autre part, les résultats identifient le rappel comme une phase bien plus dynamique que présumée, et suggèrent que l’expression de la peur conditionnée mette en jeu la régulation du trafic des récepteurs AMPA par les récepteurs NMDA. Le présent travail espère contribuer à la compréhension de la neurobiologie fondamentale de la mémoire. De plus, il propose une intégration des résultats aux modèles animaux d’étude des troubles psychologiques conséquents aux mémoires traumatiques chez l’humain, tels que les phobies et les syndromes de stress post-traumatiques. / Synaptic plasticity is necessary for the acquisition of memory in all studied species, from invertebrates to primates. Memory formation starts with a phase of plasticity that entails synaptic remodeling ; then follows the consolidation of these modifications, which contributes to long-term memory. Some memories return to a malleable state upon retrieval. Consequently, the memory trace enters a new phase of plasticity, during which some memory components are eventually updated, then reconsolidated. The aim of the present thesis was to study the cellular and molecular mechanisms that are engaged during memory retrieval. We used a model of Pavlovian conditioning in Sprague Dawley rats, combined to pharmacological manipulations and quantitative analysis of synaptic plasticity markers, in order to study the dynamics of auditory fear memory. The neuronal circuitry and the associative mechanisms involved in the neurobiology of this memory are well characterized, in particular the role of NMDA and AMPA glutamatergic receptors in synaptic plasticity and consolidation. Our results show that the return of the memory trace to lability requires activation of NMDA receptors in the basolateral amygdala during retrieval, whereas AMPA receptors are necessary for the behavioral expression of the conditioned fear response. Furthermore, the data identify retrieval as being much more dynamic than recognized, and suggest that conditioned fear expression involves NMDA receptor-dependent regulation of AMPA receptors’ trafficking. The present work attempts to advance our understanding of the fundamental neurobiology of memory. In addition, it offers an integrative view of the data with regards to animal modeling of human clinical issues related to traumatic memories, like phobias and post-traumatic stress disorders.

O estudo dos mecanismos neurobiológicos associados a memórias aversivas tem aplicações potenciais à prevenção e ao tratamento de patologias psiquiátricas associadas a memórias traumáticas. Em particular, o processo de reconsolidação, através do qual memórias evocadas são novamente estabilizadas, tem sido visto como um possível mecanismo na origem desses transtornos e, ao mesmo tempo, um alvo para estratégias terapêuticas. Experiências precoces influenciam o desenvolvimento e a maturação de circuitos encefálicos, e a sua interação com a carga genética do indivíduo, pode influenciar as estratégias de enfrentamento de situações aversivas ao longo da vida, gerando indivíduos resilientes ou vulneráveis ao desenvolvimento de transtornos psiquiátricos. Dentro deste contexto, o objetivo principal desta tese foi estudar diferentes experiências precoces, em modelos animais, e seu efeito sobre o processo de reconsolidação de memórias aversivas e, consequentemente, sobre o desenvolvimento de padrões de conduta de resiliência ou vulnerabilidade. Foram utilizados dois modelos, em ratos: a manipulação neonatal e a separação materna. Os resultados obtidos mostram que ambas intervenções levaram a um aumento dos cuidados que a mãe dedica à prole, mas a separação materna induziu um comportamento mais inconsistente, que reflete menor qualidade; consistentemente, a prole de ambos os sexos exibiu alterações na secreção de corticosterona frente a um contexto previamente pareado com um estímulo aversivo, e os machos mostraram generalização do medo a um contexto novo após a experiência. A manipulação neonatal, além de gerar um aumento do cuidado materno, levou a um aumento dos níveis centrais de ocitocina nas mães. A prole do sexo masculino exibiu comportamento de congelamento diminuído no contexto condicionado a um estímulo aversivo. Ambas intervenções geraram resistência à reconsolidação da memória aversiva condicionada, através de um mecanismo que parece envolver o hipocampo dorsal mas não a amígdala basolateral; o hipocampo ventral de ratos machos separados no período neonatal mostrou uma diminuição de espécies reativas de oxigênio e nitrogênio, sugestivo de atenuação de alguns mecanismos de plasticidade. Estes resultados apontam que os padrões de comportamento frente a situações aversivas são afetados pelas experiências neonatais, como reportado anteriormente, e que a manipulação parece gerar uma conduta mais resiliente enquanto a separação está associada ao surgimento de um padrão de comportamento mais vulnerável. A reconsolidação de memória de medo alterada em conjunto com a generalização da memória e a inconsistência comportamental da mãe encontrados neste trabalho e as alterações estruturais e funcionais na amígdala nestes animais, reportadas anteriormente, tornam a separação materna um modelo promissor para o estudo de mecanismos neurobiológicos dos transtornos psiquiátricos associados a memórias traumáticas. / The prevention and treatment of psychiatric pathologies associated with traumatic memories benefits from the study of the neurobiological mechanisms underlying aversive memories. In particular, the reconsolidation process, through which retrieved memories are restabilized, has been regarded both as a possible mechanism at the origin of these disorders and as a target for therapeutical strategies. Early life experiences impact the development and maturation of brain circuits, and their interaction with the individual’s genetic load may influence the coping mechanisms throughout life, generating individuals that are resilient or vulnerable to psychiatric disorders. Therefore, the aim of this thesis was to study, in an animal model, different early interventions and their effect on fear memory reconsolidation, and consequently the development of behavioral patterns of resilience or vulnerability. Neonatal handling and maternal separation in rats were used as early intervention models. The results obtained show that both interventions led to an increase in the amount of care the dam provides to the offspring, but maternal separation increased behavioral inconsistency, which reflects low quality behavior; consistently, both male and female offspring exhibited changes in corticosterone secretion in response to a context that was previously paired with an aversive stimulus and males showed fear generalization to a novel context after the experience. Neonatal handling increased both maternal care and central oxytocin levels in the dam. The male offspring showed reduced freezing behavior in the context conditioned to an aversive stimulus. Both interventions generated resistance to conditioned fear memory reconsolidation, through a mechanism that appears to involve the dorsal hippocampus but not the basolateral amygdala; the ventral hippocampus of males that were separated in the neonatal period had decreased reactive oxygen and nitrogen species, suggesting attenuated plasticity mechanisms. These results suggest that behavioral patterns which emerge when facing aversive situations are affected by neonatal experiences, as reported previously, and that neonatal handling appears to result in resilience while maternal separation appears to be associated with a pattern of vulnerability. Changes in fear memory reconsolidation together with memory generalization and maternal behavior inconsistency, found in this work, and the structural and functional changes in the amygdala, reported earlier, suggest that maternal separation is a promising model to study the neurobiological mechanisms of psychiatric pathologies associated with traumatic memories.

Estudos sobre extinção do condicionamento Pavloviano envolvendo estímulos aversivos (condicionamento de medo) são considerados como análogos experimentais das terapias por exposição, nas quais pacientes são confrontados com situações temidas (porém seguras) com o objetivo de reduzir respostas de medo. Nos experimentos sobre extinção, estímulos que eliciam respostas condicionais (estímulos condicionais, CSs) por terem sido previamente associados a estímulos aversivos incondicionais (estímulos incondicionais, US) são apresentados repetidamente na ausência do US. Como resultado, as respostas condicionais de medo diminuem. Um desafio nessa área é sustentar a redução do medo a longo prazo, visto que o retorno de respostas condicionais (retorno do medo) é comumente observado no laboratório e na clínica. Estudos recentes conseguiram impedir o retorno do medo por meio da extinção pós-recuperação (post-retrieval extinction, PRE), procedimento que consiste em extinção após a apresentação de um estímulo que estava presente durante o condicionamento (retrieval cue). Contudo, tentativas de replicação desse procedimento geraram resultados conflitantes. O objetivo desta tese é contribuir para o debate sobre as variáveis envolvidas no retorno do medo com o uso da PRE em humanos. Um experimento foi conduzido para verificar se o comportamento verbal emitido pelos participantes durante a PRE pode mudar a probabilidade de retorno do medo. Participantes adultos (n=57) foram submetidos a condicionamento Pavloviano diferencial no qual uma fotografia de uma face humana (CS+) foi pareada a um estímulo elétrico leve (US), enquanto que outra fotografia de face humana nunca foi pareada ao US. No dia seguinte, os participantes foram alocados em um de três grupos (n=19): Experimental atividade verbal relacionada (Exp R), Experimental atividade verbal não-relacionada (Exp N) e Controle. Todos os grupos passaram por extinção, mas para os grupos experimentais, esse procedimento foi antecedido em 10 min por uma pista (retrieval cue) que consistia na apresentação não-reforçada dos CSs. Durante o intervalo entre essa pista e a extinção, os participantes do grupo Exp R se engajaram numa atividade na qual tinham que fazer verbalizações relacionadas às contingências experimentais, enquanto que os participantes do grupo Exp N tinham que fazer verbalizações que não estavam relacionadas às contingências experimentais. O grupo controle foi submetido à extinção tradicional (sem apresentação de pista ou 10 min de intervalo antes da extinção). No terceiro dia, todos os participantes passaram por um teste que consistia em quatro apresentações do US seguidas de extinção (teste de restabelecimento). As respostas de condutância da pele frente ao CS e ao US foram usadas como medidas das respostas condicionais e incondicionais, respectivamente. Retorno do medo, medido pelo responder diferencial (discriminação entre CS+ e CS-) no teste, estava presente no grupo controle e em menor grau no grupo Exp R. Em comparação, sujeitos do grupo Exp N não apresentaram responder diferencial em função de diminuição nas respostas frente ao CS+ e aumento nas respostas frente ao CS-. Este estudo mostra que o comportamento verbal pode mudar os efeitos da PRE, o que tem implicações para a sua adaptação para uso clínico / Studies on extinction of Pavlovian conditioning involving aversive stimuli (fear conditioning) have been considered experimental analogues of exposure treatments in which patients are confronted with feared but safe situations in order to reduce fear responses. In extinction experiments, stimuli that elicit conditioned fear responses (conditioned stimuli, CS) because they have been previously associated with aversive stimuli (unconditioned stimuli, US) are repeatedly presented in the absence of the US. As a result, conditioned fear responses tend to diminish. The challenge in this area is how to maintain fear reduction in the long term, as return of conditioned fear responses (return of fear) is commonly observed in laboratory and clinical settings. Recent studies were able to prevent return of fear by means of post-retrieval extinction (PRE), a procedure consisting of extinction after the presentation of a stimulus that was present during conditioning (retrieval cue). However, replications of this procedure have yielded mixed results. With this thesis, I attempted to contribute to the debate on the variables that determine the probability of return of fear after PRE in humans. An experiment was conducted to test if verbal behavior emitted by participants during PRE can change the probability of return of fear. Adult participants (n=57) underwent differential Pavlovian conditioning in which one photograph of a human face (CS+) was paired with a mild electrical stimulus (US), whereas another photograph of human face was not paired with the US. On the next day, participants were designated to one of three groups (n=19): Experimental related verbal activity (Exp R), Experimental non-related verbal activity (Exp N), and Control. All groups underwent extinction but for experimental groups, a retrieval cue consisting of a single unreinforced presentation of the CSs was carried out 10-min prior to extinction. During the interval between retrieval cue and extinction, participants from the Exp R group were required to engage in an activity directing their overt verbal behavior towards the experimental contingencies, whereas participants from the Exp N group were required to engage in an activity directing their overt verbal behavior away from the experimental contingencies. Control group underwent a standard extinction procedure (no retrieval cue or 10-min interval prior to extinction). On a third day, all participants underwent a test consisting of four presentations of the US alone followed by extinction (reinstatement test). Skin conductance responses to the presentations of the CSs and US were used as the dependent measure of conditioned and unconditioned responses, respectively. Return of fear, as measured through differential responding (discrimination between CS+ and CS-), was present in subjects from the control group and to a lesser extent in subjects from the Exp R group. In contrast, differential responding was abolished in subjects from the Exp N group, a result that was dependent both on decrease in responses to the CS+ as well as increase in responses to the CS-. This study shows that verbal behavior might change the effects of PRE, which can have implication for its adaptation for treating pathological fear

Estudos sobre extinção do condicionamento Pavloviano envolvendo estímulos aversivos (condicionamento de medo) são considerados como análogos experimentais das terapias por exposição, nas quais pacientes são confrontados com situações temidas (porém seguras) com o objetivo de reduzir respostas de medo. Nos experimentos sobre extinção, estímulos que eliciam respostas condicionais (estímulos condicionais, CSs) por terem sido previamente associados a estímulos aversivos incondicionais (estímulos incondicionais, US) são apresentados repetidamente na ausência do US. Como resultado, as respostas condicionais de medo diminuem. Um desafio nessa área é sustentar a redução do medo a longo prazo, visto que o retorno de respostas condicionais (retorno do medo) é comumente observado no laboratório e na clínica. Estudos recentes conseguiram impedir o retorno do medo por meio da extinção pós-recuperação (post-retrieval extinction, PRE), procedimento que consiste em extinção após a apresentação de um estímulo que estava presente durante o condicionamento (retrieval cue). Contudo, tentativas de replicação desse procedimento geraram resultados conflitantes. O objetivo desta tese é contribuir para o debate sobre as variáveis envolvidas no retorno do medo com o uso da PRE em humanos. Um experimento foi conduzido para verificar se o comportamento verbal emitido pelos participantes durante a PRE pode mudar a probabilidade de retorno do medo. Participantes adultos (n=57) foram submetidos a condicionamento Pavloviano diferencial no qual uma fotografia de uma face humana (CS+) foi pareada a um estímulo elétrico leve (US), enquanto que outra fotografia de face humana nunca foi pareada ao US. No dia seguinte, os participantes foram alocados em um de três grupos (n=19): Experimental atividade verbal relacionada (Exp R), Experimental atividade verbal não-relacionada (Exp N) e Controle. Todos os grupos passaram por extinção, mas para os grupos experimentais, esse procedimento foi antecedido em 10 min por uma pista (retrieval cue) que consistia na apresentação não-reforçada dos CSs. Durante o intervalo entre essa pista e a extinção, os participantes do grupo Exp R se engajaram numa atividade na qual tinham que fazer verbalizações relacionadas às contingências experimentais, enquanto que os participantes do grupo Exp N tinham que fazer verbalizações que não estavam relacionadas às contingências experimentais. O grupo controle foi submetido à extinção tradicional (sem apresentação de pista ou 10 min de intervalo antes da extinção). No terceiro dia, todos os participantes passaram por um teste que consistia em quatro apresentações do US seguidas de extinção (teste de restabelecimento). As respostas de condutância da pele frente ao CS e ao US foram usadas como medidas das respostas condicionais e incondicionais, respectivamente. Retorno do medo, medido pelo responder diferencial (discriminação entre CS+ e CS-) no teste, estava presente no grupo controle e em menor grau no grupo Exp R. Em comparação, sujeitos do grupo Exp N não apresentaram responder diferencial em função de diminuição nas respostas frente ao CS+ e aumento nas respostas frente ao CS-. Este estudo mostra que o comportamento verbal pode mudar os efeitos da PRE, o que tem implicações para a sua adaptação para uso clínico / Studies on extinction of Pavlovian conditioning involving aversive stimuli (fear conditioning) have been considered experimental analogues of exposure treatments in which patients are confronted with feared but safe situations in order to reduce fear responses. In extinction experiments, stimuli that elicit conditioned fear responses (conditioned stimuli, CS) because they have been previously associated with aversive stimuli (unconditioned stimuli, US) are repeatedly presented in the absence of the US. As a result, conditioned fear responses tend to diminish. The challenge in this area is how to maintain fear reduction in the long term, as return of conditioned fear responses (return of fear) is commonly observed in laboratory and clinical settings. Recent studies were able to prevent return of fear by means of post-retrieval extinction (PRE), a procedure consisting of extinction after the presentation of a stimulus that was present during conditioning (retrieval cue). However, replications of this procedure have yielded mixed results. With this thesis, I attempted to contribute to the debate on the variables that determine the probability of return of fear after PRE in humans. An experiment was conducted to test if verbal behavior emitted by participants during PRE can change the probability of return of fear. Adult participants (n=57) underwent differential Pavlovian conditioning in which one photograph of a human face (CS+) was paired with a mild electrical stimulus (US), whereas another photograph of human face was not paired with the US. On the next day, participants were designated to one of three groups (n=19): Experimental related verbal activity (Exp R), Experimental non-related verbal activity (Exp N), and Control. All groups underwent extinction but for experimental groups, a retrieval cue consisting of a single unreinforced presentation of the CSs was carried out 10-min prior to extinction. During the interval between retrieval cue and extinction, participants from the Exp R group were required to engage in an activity directing their overt verbal behavior towards the experimental contingencies, whereas participants from the Exp N group were required to engage in an activity directing their overt verbal behavior away from the experimental contingencies. Control group underwent a standard extinction procedure (no retrieval cue or 10-min interval prior to extinction). On a third day, all participants underwent a test consisting of four presentations of the US alone followed by extinction (reinstatement test). Skin conductance responses to the presentations of the CSs and US were used as the dependent measure of conditioned and unconditioned responses, respectively. Return of fear, as measured through differential responding (discrimination between CS+ and CS-), was present in subjects from the control group and to a lesser extent in subjects from the Exp R group. In contrast, differential responding was abolished in subjects from the Exp N group, a result that was dependent both on decrease in responses to the CS+ as well as increase in responses to the CS-. This study shows that verbal behavior might change the effects of PRE, which can have implication for its adaptation for treating pathological fear

Memory stabilization following encoding (synaptic consolidation) or memory reactivation (reconsolidation) requires gene expression and protein synthesis. Although consolidation and reconsolidation may be mediated by distinct molecular mechanisms, disrupting the function of the transcription factor CREB (cAMP responsive element binding protein) impairs both processes. We use a gain-of-function approach to show that CREB (and CREB-coactivator CRTC1) can facilitate both synaptic and systems consolidation and reconsolidation. We first examine whether acutely increasing CREB levels in the dorsal hippocampus is sufficient to enhance spatial memory formation in the watermaze. Locally and acutely increasing CREB in the dorsal hippocampus using viral vectors is sufficient to induce robust spatial memory in two conditions which do not normally support consolidation, weakly-trained wild-type (WT) mice and strongly-trained mutant mice with brain-wide disrupted CREB function. CRTCs (CREB regulated transcription co-activators) are a powerful co-activator of CREB, but their role in memory is virtually unexplored. We show, for the first time, that the novel CREB co-activator CRTC1 enhances memory consolidation. Locally increasing CRTC1 (or CREB) in the dorsal hippocampus of WT mice prior to weak context fear conditioning facilitates consolidation of precise context memory. Last, we show that CREB or CRTC1 facilitates precise and enduring memory consolidation and reconsolidation. Acute enhancement of hippocampal CREB or CRTC1 during initial synaptic consolidation can maintain precision of remote context memory, while increasing CREB or CRTC1 just prior to reactivation of a weak remote context memory enhances context memory reconsolidation. These gain-of-function manipulations indicate that increasing CRTC1 or CREB function is sufficient to enhance the strength of new, as well as reactivated established, memories without compromising memory specificity. Together with previous results, these findings indicate that CREB is both necessary and sufficient for hippocampal-dependent memory formation, and underline its pivotal role in the hippocampal molecular machinery underlying long-term memory consolidation and reconsolidation.

Memory stabilization following encoding (synaptic consolidation) or memory reactivation (reconsolidation) requires gene expression and protein synthesis. Although consolidation and reconsolidation may be mediated by distinct molecular mechanisms, disrupting the function of the transcription factor CREB (cAMP responsive element binding protein) impairs both processes. We use a gain-of-function approach to show that CREB (and CREB-coactivator CRTC1) can facilitate both synaptic and systems consolidation and reconsolidation. We first examine whether acutely increasing CREB levels in the dorsal hippocampus is sufficient to enhance spatial memory formation in the watermaze. Locally and acutely increasing CREB in the dorsal hippocampus using viral vectors is sufficient to induce robust spatial memory in two conditions which do not normally support consolidation, weakly-trained wild-type (WT) mice and strongly-trained mutant mice with brain-wide disrupted CREB function. CRTCs (CREB regulated transcription co-activators) are a powerful co-activator of CREB, but their role in memory is virtually unexplored. We show, for the first time, that the novel CREB co-activator CRTC1 enhances memory consolidation. Locally increasing CRTC1 (or CREB) in the dorsal hippocampus of WT mice prior to weak context fear conditioning facilitates consolidation of precise context memory. Last, we show that CREB or CRTC1 facilitates precise and enduring memory consolidation and reconsolidation. Acute enhancement of hippocampal CREB or CRTC1 during initial synaptic consolidation can maintain precision of remote context memory, while increasing CREB or CRTC1 just prior to reactivation of a weak remote context memory enhances context memory reconsolidation. These gain-of-function manipulations indicate that increasing CRTC1 or CREB function is sufficient to enhance the strength of new, as well as reactivated established, memories without compromising memory specificity. Together with previous results, these findings indicate that CREB is both necessary and sufficient for hippocampal-dependent memory formation, and underline its pivotal role in the hippocampal molecular machinery underlying long-term memory consolidation and reconsolidation.

O estudo dos mecanismos neurobiológicos associados a memórias aversivas tem aplicações potenciais à prevenção e ao tratamento de patologias psiquiátricas associadas a memórias traumáticas. Em particular, o processo de reconsolidação, através do qual memórias evocadas são novamente estabilizadas, tem sido visto como um possível mecanismo na origem desses transtornos e, ao mesmo tempo, um alvo para estratégias terapêuticas. Experiências precoces influenciam o desenvolvimento e a maturação de circuitos encefálicos, e a sua interação com a carga genética do indivíduo, pode influenciar as estratégias de enfrentamento de situações aversivas ao longo da vida, gerando indivíduos resilientes ou vulneráveis ao desenvolvimento de transtornos psiquiátricos. Dentro deste contexto, o objetivo principal desta tese foi estudar diferentes experiências precoces, em modelos animais, e seu efeito sobre o processo de reconsolidação de memórias aversivas e, consequentemente, sobre o desenvolvimento de padrões de conduta de resiliência ou vulnerabilidade. Foram utilizados dois modelos, em ratos: a manipulação neonatal e a separação materna. Os resultados obtidos mostram que ambas intervenções levaram a um aumento dos cuidados que a mãe dedica à prole, mas a separação materna induziu um comportamento mais inconsistente, que reflete menor qualidade; consistentemente, a prole de ambos os sexos exibiu alterações na secreção de corticosterona frente a um contexto previamente pareado com um estímulo aversivo, e os machos mostraram generalização do medo a um contexto novo após a experiência. A manipulação neonatal, além de gerar um aumento do cuidado materno, levou a um aumento dos níveis centrais de ocitocina nas mães. A prole do sexo masculino exibiu comportamento de congelamento diminuído no contexto condicionado a um estímulo aversivo. Ambas intervenções geraram resistência à reconsolidação da memória aversiva condicionada, através de um mecanismo que parece envolver o hipocampo dorsal mas não a amígdala basolateral; o hipocampo ventral de ratos machos separados no período neonatal mostrou uma diminuição de espécies reativas de oxigênio e nitrogênio, sugestivo de atenuação de alguns mecanismos de plasticidade. Estes resultados apontam que os padrões de comportamento frente a situações aversivas são afetados pelas experiências neonatais, como reportado anteriormente, e que a manipulação parece gerar uma conduta mais resiliente enquanto a separação está associada ao surgimento de um padrão de comportamento mais vulnerável. A reconsolidação de memória de medo alterada em conjunto com a generalização da memória e a inconsistência comportamental da mãe encontrados neste trabalho e as alterações estruturais e funcionais na amígdala nestes animais, reportadas anteriormente, tornam a separação materna um modelo promissor para o estudo de mecanismos neurobiológicos dos transtornos psiquiátricos associados a memórias traumáticas. / The prevention and treatment of psychiatric pathologies associated with traumatic memories benefits from the study of the neurobiological mechanisms underlying aversive memories. In particular, the reconsolidation process, through which retrieved memories are restabilized, has been regarded both as a possible mechanism at the origin of these disorders and as a target for therapeutical strategies. Early life experiences impact the development and maturation of brain circuits, and their interaction with the individual’s genetic load may influence the coping mechanisms throughout life, generating individuals that are resilient or vulnerable to psychiatric disorders. Therefore, the aim of this thesis was to study, in an animal model, different early interventions and their effect on fear memory reconsolidation, and consequently the development of behavioral patterns of resilience or vulnerability. Neonatal handling and maternal separation in rats were used as early intervention models. The results obtained show that both interventions led to an increase in the amount of care the dam provides to the offspring, but maternal separation increased behavioral inconsistency, which reflects low quality behavior; consistently, both male and female offspring exhibited changes in corticosterone secretion in response to a context that was previously paired with an aversive stimulus and males showed fear generalization to a novel context after the experience. Neonatal handling increased both maternal care and central oxytocin levels in the dam. The male offspring showed reduced freezing behavior in the context conditioned to an aversive stimulus. Both interventions generated resistance to conditioned fear memory reconsolidation, through a mechanism that appears to involve the dorsal hippocampus but not the basolateral amygdala; the ventral hippocampus of males that were separated in the neonatal period had decreased reactive oxygen and nitrogen species, suggesting attenuated plasticity mechanisms. These results suggest that behavioral patterns which emerge when facing aversive situations are affected by neonatal experiences, as reported previously, and that neonatal handling appears to result in resilience while maternal separation appears to be associated with a pattern of vulnerability. Changes in fear memory reconsolidation together with memory generalization and maternal behavior inconsistency, found in this work, and the structural and functional changes in the amygdala, reported earlier, suggest that maternal separation is a promising model to study the neurobiological mechanisms of psychiatric pathologies associated with traumatic memories.

O estudo dos mecanismos neurobiológicos associados a memórias aversivas tem aplicações potenciais à prevenção e ao tratamento de patologias psiquiátricas associadas a memórias traumáticas. Em particular, o processo de reconsolidação, através do qual memórias evocadas são novamente estabilizadas, tem sido visto como um possível mecanismo na origem desses transtornos e, ao mesmo tempo, um alvo para estratégias terapêuticas. Experiências precoces influenciam o desenvolvimento e a maturação de circuitos encefálicos, e a sua interação com a carga genética do indivíduo, pode influenciar as estratégias de enfrentamento de situações aversivas ao longo da vida, gerando indivíduos resilientes ou vulneráveis ao desenvolvimento de transtornos psiquiátricos. Dentro deste contexto, o objetivo principal desta tese foi estudar diferentes experiências precoces, em modelos animais, e seu efeito sobre o processo de reconsolidação de memórias aversivas e, consequentemente, sobre o desenvolvimento de padrões de conduta de resiliência ou vulnerabilidade. Foram utilizados dois modelos, em ratos: a manipulação neonatal e a separação materna. Os resultados obtidos mostram que ambas intervenções levaram a um aumento dos cuidados que a mãe dedica à prole, mas a separação materna induziu um comportamento mais inconsistente, que reflete menor qualidade; consistentemente, a prole de ambos os sexos exibiu alterações na secreção de corticosterona frente a um contexto previamente pareado com um estímulo aversivo, e os machos mostraram generalização do medo a um contexto novo após a experiência. A manipulação neonatal, além de gerar um aumento do cuidado materno, levou a um aumento dos níveis centrais de ocitocina nas mães. A prole do sexo masculino exibiu comportamento de congelamento diminuído no contexto condicionado a um estímulo aversivo. Ambas intervenções geraram resistência à reconsolidação da memória aversiva condicionada, através de um mecanismo que parece envolver o hipocampo dorsal mas não a amígdala basolateral; o hipocampo ventral de ratos machos separados no período neonatal mostrou uma diminuição de espécies reativas de oxigênio e nitrogênio, sugestivo de atenuação de alguns mecanismos de plasticidade. Estes resultados apontam que os padrões de comportamento frente a situações aversivas são afetados pelas experiências neonatais, como reportado anteriormente, e que a manipulação parece gerar uma conduta mais resiliente enquanto a separação está associada ao surgimento de um padrão de comportamento mais vulnerável. A reconsolidação de memória de medo alterada em conjunto com a generalização da memória e a inconsistência comportamental da mãe encontrados neste trabalho e as alterações estruturais e funcionais na amígdala nestes animais, reportadas anteriormente, tornam a separação materna um modelo promissor para o estudo de mecanismos neurobiológicos dos transtornos psiquiátricos associados a memórias traumáticas. / The prevention and treatment of psychiatric pathologies associated with traumatic memories benefits from the study of the neurobiological mechanisms underlying aversive memories. In particular, the reconsolidation process, through which retrieved memories are restabilized, has been regarded both as a possible mechanism at the origin of these disorders and as a target for therapeutical strategies. Early life experiences impact the development and maturation of brain circuits, and their interaction with the individual’s genetic load may influence the coping mechanisms throughout life, generating individuals that are resilient or vulnerable to psychiatric disorders. Therefore, the aim of this thesis was to study, in an animal model, different early interventions and their effect on fear memory reconsolidation, and consequently the development of behavioral patterns of resilience or vulnerability. Neonatal handling and maternal separation in rats were used as early intervention models. The results obtained show that both interventions led to an increase in the amount of care the dam provides to the offspring, but maternal separation increased behavioral inconsistency, which reflects low quality behavior; consistently, both male and female offspring exhibited changes in corticosterone secretion in response to a context that was previously paired with an aversive stimulus and males showed fear generalization to a novel context after the experience. Neonatal handling increased both maternal care and central oxytocin levels in the dam. The male offspring showed reduced freezing behavior in the context conditioned to an aversive stimulus. Both interventions generated resistance to conditioned fear memory reconsolidation, through a mechanism that appears to involve the dorsal hippocampus but not the basolateral amygdala; the ventral hippocampus of males that were separated in the neonatal period had decreased reactive oxygen and nitrogen species, suggesting attenuated plasticity mechanisms. These results suggest that behavioral patterns which emerge when facing aversive situations are affected by neonatal experiences, as reported previously, and that neonatal handling appears to result in resilience while maternal separation appears to be associated with a pattern of vulnerability. Changes in fear memory reconsolidation together with memory generalization and maternal behavior inconsistency, found in this work, and the structural and functional changes in the amygdala, reported earlier, suggest that maternal separation is a promising model to study the neurobiological mechanisms of psychiatric pathologies associated with traumatic memories.

La présente thèse porte sur l’évaluation de l’efficacité d’un nouveau traitement pour le trouble de stress post-traumatique (TSPT). Le traitement a été développé selon les prémisses de la théorie de la reconsolidation des souvenirs. Il consiste en six courtes séances de remémoration de l’événement traumatique réalisées sous l’effet du propranolol, un bêtabloquant. La population de l’étude est constituée de patients souffrant d’un TSPT chronique. La thèse comporte cinq chapitres. Le premier chapitre est l’introduction, on y retrouve une description du TSPT, des traitements validés empiriquement, de diverses théories de la mémoire, d’un modèle étiologique du TSPT, d’études sur la consolidation et la reconsolidation, de la pharmacocinétique et du mécanisme d’action du propranolol,ainsi que des objectifs de la thèse. Le second chapitre est une revue critique de littérature sur la théorie de la reconsolidation. Comme l’étude du phénomène de la reconsolidation est récente, nous tentons de faire le point sur l’état des connaissances dans le domaine, dans un effort de réflexion sur la validité de la théorie. Nous proposons une série de critères permettant de différencier la reconsolidation d’autres processus connexes. Nous concluons que la théorie paraît valide, bien que d’autres études soient nécessaires afin de rendre compte de résultats négatifs publiés par le passé. Le troisième chapitre est un essai ouvert, et vise à évaluer l’efficacité d’un traitement basé sur la reconsolidation à diminuer la sévérité et l’incidence du TSPT, auprès de 42 patients souffrant d’un TSPT chronique. Le traitement consiste en six séances de remémoration de l’événement traumatique sous propranolol. Lors d’un suivi à trois mois, nous rapportons une diminution des symptômes de TSPT de 41%-56%, ainsi qu’une diminution de l’incidence du TSPT de 74%. En comparaison, seulement 2/25 patients du groupe contrôle (ayant participé uniquement aux évaluations) ne souffrent plus d’un TSPT. Dans le groupe traitement, les tailles d’effet (d de Cohen)varient entre 1.32-2.19. Le quatrième chapitre a comme objectif d’identifier des caractéristiques des patients prédisant l’efficacité du traitement, et d’explorer s’ils s’améliorent dans des domaines de santé autres que le TSPT. Nous rapportons que les femmes s’améliorent davantage que les hommes, mais que d’autres facteurs, tels que la sévérité des traits de personnalité borderline ou le type de trauma (enfance versus adulte), n’influent pas sur l’efficacité. Également, les patients s’améliorent dans les domaines de santé suivants : la qualité de vie, la symptomatologie dépressive, l’intensité des émotions négatives au rappel de l’événement traumatique et dans la vie courante. Le cinquième chapitre contient la discussion générale de la thèse. Nous effectuons une synthèse et interprétation des résultats, nous examinons les hypothèses alternatives à l’amélioration clinique et abordons des pistes de recherches futures. Nous concluons que le traitement à l’étude a été efficace dans notre échantillon de patients souffrant d’un TSPT chronique. Étant donné la méthodologie employée (essai ouvert), nous ne pouvons statuer sur le mécanisme d’action du traitement, à savoir si l’amélioration clinique a été réellement causée par un blocage de la reconsolidation des souvenirs. / This dissertation aims at exploring the efficacy of a new treatment for posttraumatic stress disorder (PTSD). The treatment was developed in accordance with an emerging theory in neuroscience, the reconsolidation theory, and it consists in six short reactivation sessions of a traumatic memory under the influence of propranolol (a ß-blocker), with patients suffering from longstanding PTSD. This dissertation includes five chapters. Chapter I is the introduction, it includes a discussion on the following topics : definition and prevalence of PTSD,empirically validated treatments in the field, memory theories, etiology of PTSD, studies on consolidation and reconsolidation, pharmacokinetics of propranolol and its mechanism of action in reconsolidation, and the objectives of the dissertation. Chapter II is a critical literature review on reconsolidation theory. We discuss some of the contradicting findings in reconsolidation, as some researchers have reported negative results in the field. We address the possibility to reconcile these discrepancies,within the scope of evaluating the validity of the theory. We also discuss a series of criterion which could provide guidance in differentiating reconsolidation from other processes. We conclude that reconsolidation theory seems valid, although more research is needed in order to shed light on some negative results that were published in the past. Chapter III is an open label trial comprising six sessions of treatment (trauma reactivation under propranolol) with 42 patients suffering from chronic PTSD. At a three-month follow-up, we report that patients have a 41% - 56% reduction in PTSD symptoms, and that 31 / 42 patients no longer meet the diagnostic threshold for PTSD. In comparison, only 2 / 25 patients from the control group (assessments only) don’t meet the diagnostic threshold for PTSD. In the treatment group, effect sizes (Cohen’s d) range between 1.32 -2.19. Chapter IV follows on the previous chapter’s study, and aims at identifying predictors of treatment outcome (i.e., predictors of the improvement in PTSD symptoms), and whether patients also improve in health domains other than PTSD. We report that women improve more than men during the treatment, but that other factors such as borderline personality severity traits or type of trauma (childhood versus adulthood) do not influence treatment outcome. Patients also improve in diverse health domains during the treatment; they have a better quality of life, less depressive symptoms, less intense negative emotions in daily life and during trauma recollection. Chapter V contains a general discussion and a conclusion. We summarize and interpret the results, we explore alternative hypotheses to the clinical improvement as well as future research directions. We conclude that this treatment yielded interesting results in our sample of patients suffering from chronic PTSD. However, our methodology (open label study) doesn’t provide any information on the mechanism of action of the treatment used in this dissertation, i.e. whether the clinical improvement was caused or not by reconsolidation blockade.

Une récente théorie de la mémoire a proposé que lorsqu'un souvenir déjà bien consolidé est réactivé, il redevient labile et susceptible aux modifications avant d'être restabilisé (reconsolidé) en mémoire à long terme. Ce nouveau modèle réfute le modèle classique de la consolidation qui propose qu'une fois consolidés, les souvenirs soient permanents et donc résistants aux effets des agents amnésiques. Les études validant la théorie de la reconsolidation abondent chez les animaux, mais encore peu d'études ont été réalisées chez les humains. L'objectif de cette thèse est de vérifier, dans une population de sujets sains et de sujets souffrant de trouble de stress post-traumatique (TSPT), l'efficacité d'un agent pharmacologique, le propranolol (un β-bloquant noradrénergique) à atténuer des souvenirs émotionnels nouvellement acquis ou déjà bien consolidés. Plus spécifiquement, nous avons mené un essai clinique contrôlé à double insu chez des sujets sains en leur administrant du propranolol (vs du placebo) lors de l'acquisition d'un nouveau souvenir et une semaine plus tard, lors de sa réactivation. L'objectif du premier article était d'évaluer l'efficacité du propranolol à diminuer la consolidation et la reconsolidation d'un souvenir émotionnel. Par ailleurs, puisque les études chez les animaux ont démontré que ces deux processus mnésiques s'effectuent à l'intérieur d'une fenêtre temporelle précise, le moment de l'administration du propranolol fut pris en considération. Les résultats ont démontré que le propranolol est en mesure de diminuer la consolidation et la reconsolidation d'une histoire émotionnelle tel que démontré par un pourcentage de bonnes réponses plus faible que le groupe contrôle lors des rappels. Toutefois, pour que cet effet soit observé, le propranolol doit être administré une heure avant la présentation des stimuli, pour la consolidation et une heure avant leur réactivation, pour la reconsolidation. En outre, les études portant sur la consolidation et la reconsolidation chez les animaux et chez les humains obtiennent parfois des résultats contradictoires. Ceci pourrait s'expliquer par le type de stimuli utilisé. Ainsi, l'objectif du second article était de préciser quel type d'information est le plus susceptible d'être affecté par le propranolol lors de son acquisition (consolidation) et lors de sa réactivation (reconsolidation). Pour ce faire, les éléments de l'histoire émotionnelle ont été divisés en fonction de leur valence (émotionnel ou neutre) et de leur centralité (central ou périphérique). Les résultats ont démontré le propranolol affecte l'ensemble des informations centrales lors du blocage de la consolidation, mais qu'il affecte plus spécifiquement les éléments émotionnels centraux lors de la reconsolidation. Notre groupe ayant précédemment démontré que le traitement avec le propranolol est en mesure de réduire les symptômes de TSPT chez une population clinique, nous nous sommes interrogés sur son efficacité à diminuer la mémoire implicite d'un événement traumatique. Le propranolol a été administré aux participants à 6 reprises (une fois par semaine sur une période de 6 semaines) lors de la réactivation de leur trauma. Les résultats ont révélé que le traitement avec le propranolol est en mesure de diminuer la réponse psychophysiologique des participants à l'écoute du compte rendu de leur trauma une semaine et 4 mois suivant la fin du traitement. En somme, cette thèse démontre que le propranolol est en mesure de bloquer la consolidation et la reconsolidation de souvenirs émotionnels chez l'humain lorsqu'il est administré une heure avant l'acquisition ou la réactivation des souvenirs. Il arrive en outre à atténuer un souvenir déclaratif émotionnel chez des sujets sains, mais également un souvenir implicite chez des sujets souffrant de TSPT. Ainsi, ces résultats ouvrent la voie à la création de nouveaux traitements pour les psychopathologies ayant comme étiologie un souvenir émotionnel intense. / A recent theory of memory proposes that when a well-consolidated memory is reactivated, it becomes labile again and susceptible to change before being restabilized (reconsolidated) in long-term memory. This new memory theory refutes the classical model of consolidation, which suggests that once consolidated, memories are permanent and hence resistant to the effects of amnestic agents. Studies validating reconsolidation theory in animals abound, but fewer studies have been conducted in humans. The objective of the current thesis is to verify the potential of the pharmacological agent propranolol (a β-blocker) to impair newly acquired or already consolidated emotional memories in healthy subjects and subjects suffering from posttraumatic stress disorder (PTSD). We conducted a double blind controlled trial where propranolol (vs. placebo) was administered to healthy subjects during the acquisition of a new memory (an emotionally valenced story) and one week later, during its reactivation. The aim of the first paper was to evaluate the efficacy of propranolol to reduce the consolidation and reconsolidation of emotional memories. Furthermore, since animal studies have shown that these two memory processes occur within a given time window, the timing of the propranolol administration was taken into consideration. The results showed that propranolol was able to dampen the consolidation and reconsolidation of an emotional story as evidenced by a lower percentage of correct answers at recall compared to the control group. However, to observe this effect, the propranolol needed to be administered one hour before the presentation of the story at consolidation and one hour before it’s reactivation at reconsolidation. While consolidation and reconsolidation studies in animals and humans have yielded conflicting results, this may depend on the type of stimuli used. The objective of the second paper of this thesis was to determine what type of information is most likely to be affected by propranolol during acquisition (consolidation) and reactivation (reconsolidation). To do this, the elements of the emotional story were divided according their emotionality (emotional vs. neutral) and centrality (central vs. peripheral). The results demonstrated that propranolol blocks the consolidation of all central information, but it affects selectively the central-emotional story elements at reconsolidation. Our group previously demonstrated that treatment with propranolol is able to reduce the symptoms of PTSD in a clinical population. In this thesis, we investigated whether it could also reduce the implicit memory of a traumatic event. Propranolol was administered on 6 different occasions (once a week over a period of 6 weeks) to participants upon reactivation of their traumatic event. The results unveiled that treatment with propranolol was able to decrease the participants’ psychophysiological responses to their traumatic script at one week post-treatment and at the 4 months follow-up. In sum, this thesis reports results suggesting that propranolol is able to block the consolidation and reconsolidation of emotional memories in humans when it is administered one hour before acquisition or before reactivation. Furthermore, this thesis suggests that propranolol impairs selectively central-emotional type of information and that it has the ability to dampen declarative emotional memory in healthy subjects, but also implicit memory in subjects suffering from PTSD. Hence, these results open the way to create new treatments for psychopathologies having at their core an intense emotional memory