We can notice the positive effects of sleep on many functions of our organism. For a long time we have observed the interconnection between sleep and memory and today we already know, that different sleep phases correlate with an improvement of different memory types. One of the hypotheses, that explain the positive effect of sleep on strengthening of the memory representations, is its irreplaceable active role in the process of memory consolidation. The memory consolidation of episodic type in animals, which processes memories into events with time and space context, could according to this theory occur due to two phase sleep process, in which each phase has a specific role. The theoretical part of this thesis will familiarize the reader with the problematic of organization of time and space in our brain, and introduce him to the foundations of electroencephalography (EEG) and offer a detailed introduction into the discussed hypothesis of active sleep consolidation. The practical part is then focused on the confirmation of the already mentioned hypothesis through the combination of comparison of results from the behavioral task of 2 groups of animals with different sleep manipulation and of the analysis of EEG signal recorded during the experiment before and after the training. The behavioral task...

La navigation et les modes de déplacement intéressent la communauté scientifique depuis maintenant près d'un demi siècle. Cependant, l’augmentation de l’incidence des troubles dégénératifs du système nerveux central chez l’homme rend plus prégnante la nécessité de compréhension de la navigation et de l’influence du temps sur celle-ci. S'il est connu chez l'homme comme chez le rongeur que l'avancée en âge affecte les capacités à se déplacer dans de vastes environnements, peu de données sont disponibles quant aux processus cognitifs impliqués dans ce type de comportement et leurs éventuelles modulations avec l'âge. La définition des stratégies utilisées, l’incidence respective des mécanismes allocentriques et égocentriques, la capacité de mise en œuvre d’une stratégie au moment demandé, lors d’un rappel à court ou à long délai, l’influence du temps qui passe sont autant de questions que nous avons abordées dans ce travail de thèse. Afin d’effectuer ces études, nous avons développé des tâches en environnements virtuels modélisés sur ordinateur et utilisé des tests neuropsychologiques nécessitant la mobilisation des compétences visuo-spatiales. Dans une première étude utilisant une épreuve de localisation spatiale, les résultats obtenus mettent en évidence chez les personnes âgées, une altération des aptitudes lors de la mise en œuvre d’une stratégie allocentrique, sans atteinte des performances égocentriques. La deuxième étude utilisant une version virtuelle du test de la piscine de Morris reconnu comme une tâche allocentrique chez le rongeur, conforte ces données. De façon similaire dans les deux études, les personnes âgées présentent une altération de la sélection et de l’exécution de la stratégie de déplacement qui s’avère optimale pour résoudre la tâche spatiale. Nous avons également mis en évidence une difficulté, chez ces mêmes participants, à utiliser une représentation mentale globale de l’espace, sans toutefois qu’il soit possible de distinguer si l’origine de cette difficulté vient d’une altération de la formation ou de la récupération de cette « carte cognitive ». Le temps pourrait également jouer son rôle de par le délai entre l'acquisition d'une information spatiale et le moment où il est nécessaire de l’utiliser à nouveau. En étudiant l’effet du délai sur la trace mnésique spatiale, nous avons observé que les sujets jeunes utilisant de façon prédominante une stratégie allocentrique voyaient leurs performances diminuer lors d’un rappel après quatre semaines alors que celles des sujets âgés restaient inchangées. Ceci soulève bien entendu la question de la différence d’encodage des informations entre les sujets jeunes et âgés, avec un versant plus détaillé chez les sujets jeunes, mais surtout s’intègre au sein du débat actuel sur l’existence d’une modification de la trace mnésique qui pourrait selon la théorie des traces multiples de la consolidation, évoluer vers un souvenir plus schématisé avec le délai. Les résultats d’une dernière étude dans laquelle nous manipulons le contexte environnemental de la piscine virtuelle de Morris, amène des arguments en faveur d’une « schématisation » du souvenir au cours de la consolidation, en mettant en évidence une absence de discrimination par les participants, d’un changement des repères spatiaux lors d’un rappel de l’information après six semaines de délai. Toutes ces données sont discutées dans le cadre du débat actuel de la consolidation, notamment sur la contribution de l’hippocampe dans le stockage et le rappel des informations anciennes. A la lumière de nos données, nous proposons une vue intégrative du fonctionnement de l’interface hippocampo-corticale lors des rappels après un court et long délai, en fonction de l’âge. / While the detrimental effects of human aging on cognitive functions are well documented, how normal aging affects spatial memory processing and the organization of recent and long-term memories remains unclear. What are the cognitive strategies used when confronted to spatial navigation in large environments? How are the selection and use of these strategies affected by aging? How are recent and long-term remote memories organized as a function of aging during systems-level consolidation? These are the questions we sought to address during the course of this Ph.D. thesis by developing a series of virtual environments aimed at assessing spatial navigation and memory performance in young adults and aged participants. In a first series of experiments, participants were tested for object location memory in a virtual environment (a medieval castle) that enabled shifts in spatial viewpoints between study and test. Aged participants exhibited poor performance relative to young adults only in the shifted view conditions, thus providing strong evidence for a decline in allocentric, but not egocentric, spatial memory. In contrast to young adults, aged participants exhibited difficulties in processing efficiently distal cues of the environment and were less prone to adopt allocentric strategies. Manipulations of the spatial layout of the environment led us to the conclusion that aging seems to preferentially interfere with the capacity to form or use mental representations built upon all pieces of the environmental features which typically, are never in full view in real world large-scale environments. In a second set of experiments, participants were tested in an ecologically-relevant virtual version of the Morris water maze which mimics that classically used in rodents. Aged participants performed more poorly compared to middle-aged and young adults and formed a more schematic spatial memory. They favoured a directional single cue-based strategy to locate the hidden platform contrasting with young adults who formed complex geometrical relationships between distal cues of the environment. A neuropsychological test battery confirmed that binding of unrelated items and abilities to mentally manipulate information were two processes involved in solving the water maze task. Thus, upon acquisition, aged participants had difficulties in forming experientially detailed cognitive maps and in binding unrelated features of the environment into a cohesive spatial memory, possibly indicative of altered hippocampal-frontal circuitry. We next proceeded to examine the organization of spatial memory as a function of time. Long-term memory assessed 4 weeks after acquisition revealed that performance decreased more rapidly in young adults compared to elderly participants, suggesting that the passage of time differentially affects the content of spatial memory, richly detailed spatial memories being more vulnerable to decay than schematic ones. This concept of memory transformation (i.e. memories are not stored in the cortex in their original form) was supported by findings of a last experiment in which we provide evidence that participants failed in detecting changes in the spatial layout of the pool as memories matured over time. All these findings are discussed in the context of the current debate about the concept of memory consolidation which opposes the standard model of memory consolidation to the multiple trace theory, two views which make different predictions about the contribution of the hippocampus to remote memory storage and retrieval. In light of our own findings, we attempt to propose an integrative view of the functioning of the hippocampal-cortical interface during recent and remote memory retrieval as a function of normal aging.

La formation et le stockage à long terme des souvenirs mettent en jeu le processus de consolidation mnésique. S’il est maintenant bien admis que ce processus requiert une interaction entre la formation hippocampique et différentes régions corticales dépositaires des souvenirs, les mécanismes qui sous-tendent ce dialogue restent encore mal connus. En combinant chez le rat des approches comportementale, d’imagerie cellulaire et d’inactivations pharmacologiques des voies de signalisation intracérébrales impliquées dans l’épigenèse, nous avons cherché dans ce travail de thèse à élucider certains des mécanismes responsables de la formation des souvenirs au niveau cortical. Dans la perspective de pouvoir appréhender de façon temporellement précise le dialogue hippocampo-cortical au cours du processus de consolidation à l’échelle systémique, notre premier objectif a été de valider une épreuve comportementale adaptée à l’étude de ce processus, la transmission sociale de préférence alimentaire. Nos résultats ont montré que cette tâche, qui ne nécessite qu’une phase d’acquisition ponctuelle, induit une mémoire robuste et durable. Cette mémoire s’appuie sur des stimuli olfactifs de nature non spatiale. Son caractère associatif nécessite l’implication fonctionnelle de l’hippocampe et de régions corticales spécifiques comme le cortex orbitofrontal qui joue un rôle crucial dans le traitement d’informations de nature olfactive. Dans une deuxième série d’expériences, une approche d’imagerie cellulaire utilisant le facteur de transcription c-fos couplée à une approche pharmacologique d’inactivation transitoire région-spécifique a révélé le rôle crucial du cortex orbitofrontal dans le rappel d’informations anciennes (délai de 30 jours) mais pas récentes (délai de 1 jour). Nous avons par ailleurs mis en évidence que la consolidation des informations dans cette structure s’accompagnait de changements progressifs de l’architecture des réseaux neuronaux comme la formation de nouvelles synapses (synaptogénèse) ou l’augmentation du nombre d’épines dendritiques. En accord avec le modèle standard de la consolidation mnésique, ce recrutement cortical était associé à un désengagement de l’hippocampe, confirmant le rôle transitoire de cette structure dans le rappel à long terme d’informations olfactives associatives. Dans une troisième série d’expériences, nous nous sommes intéressés aux mécanismes pouvant sous-tendre l’établissement de la mémoire à long terme au niveau cortical. Nos résultats apportent un éclairage nouveau sur la dynamique des interactions hippocampo-corticales pendant la consolidation systémique en démontrant la nécessité d’un étiquetage des assemblées neuronales du cortex orbitofrontal dès l’encodage des informations. Un blocage de cet étiquetage par une inactivation de ce cortex au moment de l’interaction sociale (phase d’acquisition) a perturbé le rappel à long terme et empêché les modifications de l’architecture des réseaux neuronaux corticaux normalement associés au stockage à long terme des informations olfactives. Sur le plan cellulaire, cet étiquetage requiert l’activation des récepteurs NMDA et de la voie des MAPK, ainsi que l’acétylation des protéines histones impliquées dans la régulation de l’état transcriptionnel de la chromatine. En modifiant leur état d’acétylation, nous avons pu moduler positivement ou négativement le rappel à long terme des informations olfactives relatives à la préférence alimentaire. Ainsi, nos données soulignent l’importance du dialogue hippocampo-cortical dans l’établissement de la mémoire à long terme. / Abstract :

O estresse tem se mostrado como um agravante de diversas patologias e transtornos psiquiátricos, como o transtorno depressivo maior (TDM). Estudos apontam que o hipocampo, bem como as vias serotonérgicas presentes neste, estão envolvidas com as respostas de estresse bem como nos efeitos deletérios causados por este. O antagonismo do receptor 5-HT 7, presente nestas vias, tem apresentado um efeito antidepressivo e ansiolítico em alguns trabalhos, porém poucos são os estudos que investigam o papel do receptor 5-HT 7 nas estruturas encefálicas em relação às repostas de estresse. Diante desse dado, o presente trabalho utilizou o modelo Labirinto em Cruz Elevado (LCE) 24 horas após a restrição para investigar o papel do receptor 5-HT 7 do hipocampo na consolidação da memória aversiva. A metodologia utilizada consistiu na injeção bilateral de SB-258741, um antagonista do receptor 5-HT7, no hipocampo dorsal de ratos Wistar machos, feita logo após o estresse de restrição de movimento com exposição ao LCE 24 horas depois. Como resultado desta metodologia, foi observado um aumento na porcentagem de entradas dos animais nos braços abertos, mas não na porcentagem de tempo em que o animal permaneceu nestes braços. Este dado mostra que a injeção de um antagonista de 5-HT 7 no hipocampo leva a uma atenuação dos efeitos comportamentais da consolidação de memória aversiva, corroborando com efeito ansiolítico consequente ao antagonismo deste receptor, observado na literatura. Nesse sentido, a investigação sobre o papel desse receptor, nas respostas de estresse, pode ser útil para o desenvolvimento de novos fármacos para os tratamentos dos efeitos prejudiciais do estresse. / Stress has been shown to be an aggravating factor for various diseases and psychiatric disorders, such as major depressive disorder (MDD). Studies suggest that the hippocampus, as well as serotonergic pathways present in this structure, are involved in stress responses as well as in the deleterious effects caused by this factor. The receptor antagonism of 5-HT 7 receptor, present in these pathways, has shown an antidepressant and anxiolytic effect in some studies, but few studies are investigating the role of 5-HT 7 receptor in brain structures related to stress responses. Given this data, this study used the Elevated Plus Maze (EPM) 24 hours after the restrain stress to investigate the role of 5-HT 7 receptor in the hippocampus consolidation of aversive memory. The methodology in this study consisted of bilateral injection of SB-258741, an antagonist of the 5-HT7 receptor, into the dorsal hippocampus of male Wistar rats, made 5 minutes after the restrain stress with exposure to EPM after 24 hours. As a result of this method, an increase in the percentage of animals entries into the open arms, but not at the percentage of time the animal remained in these arms, was observed. This data shows that the injection of a 5-HT 7 antagonist in the hippocampus leads to an attenuation of the aversive memory consolidation behavioral effects, corroborating with the consequent anxiolytic effect to the antagonism of this receptor, presented in the literature. In this sense, research on the role of this receptor in stress responses may be useful for the development of new drugs for the treatment of the harmful effects of stress.

O estresse tem se mostrado como um agravante de diversas patologias e transtornos psiquiátricos, como o transtorno depressivo maior (TDM). Estudos apontam que o hipocampo, bem como as vias serotonérgicas presentes neste, estão envolvidas com as respostas de estresse bem como nos efeitos deletérios causados por este. O antagonismo do receptor 5-HT 7, presente nestas vias, tem apresentado um efeito antidepressivo e ansiolítico em alguns trabalhos, porém poucos são os estudos que investigam o papel do receptor 5-HT 7 nas estruturas encefálicas em relação às repostas de estresse. Diante desse dado, o presente trabalho utilizou o modelo Labirinto em Cruz Elevado (LCE) 24 horas após a restrição para investigar o papel do receptor 5-HT 7 do hipocampo na consolidação da memória aversiva. A metodologia utilizada consistiu na injeção bilateral de SB-258741, um antagonista do receptor 5-HT7, no hipocampo dorsal de ratos Wistar machos, feita logo após o estresse de restrição de movimento com exposição ao LCE 24 horas depois. Como resultado desta metodologia, foi observado um aumento na porcentagem de entradas dos animais nos braços abertos, mas não na porcentagem de tempo em que o animal permaneceu nestes braços. Este dado mostra que a injeção de um antagonista de 5-HT 7 no hipocampo leva a uma atenuação dos efeitos comportamentais da consolidação de memória aversiva, corroborando com efeito ansiolítico consequente ao antagonismo deste receptor, observado na literatura. Nesse sentido, a investigação sobre o papel desse receptor, nas respostas de estresse, pode ser útil para o desenvolvimento de novos fármacos para os tratamentos dos efeitos prejudiciais do estresse. / Stress has been shown to be an aggravating factor for various diseases and psychiatric disorders, such as major depressive disorder (MDD). Studies suggest that the hippocampus, as well as serotonergic pathways present in this structure, are involved in stress responses as well as in the deleterious effects caused by this factor. The receptor antagonism of 5-HT 7 receptor, present in these pathways, has shown an antidepressant and anxiolytic effect in some studies, but few studies are investigating the role of 5-HT 7 receptor in brain structures related to stress responses. Given this data, this study used the Elevated Plus Maze (EPM) 24 hours after the restrain stress to investigate the role of 5-HT 7 receptor in the hippocampus consolidation of aversive memory. The methodology in this study consisted of bilateral injection of SB-258741, an antagonist of the 5-HT7 receptor, into the dorsal hippocampus of male Wistar rats, made 5 minutes after the restrain stress with exposure to EPM after 24 hours. As a result of this method, an increase in the percentage of animals entries into the open arms, but not at the percentage of time the animal remained in these arms, was observed. This data shows that the injection of a 5-HT 7 antagonist in the hippocampus leads to an attenuation of the aversive memory consolidation behavioral effects, corroborating with the consequent anxiolytic effect to the antagonism of this receptor, presented in the literature. In this sense, research on the role of this receptor in stress responses may be useful for the development of new drugs for the treatment of the harmful effects of stress.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Caffeine, an antagonist of adenosine receptors, is one of the most widely consumed psychostimulant substance in the world. There are contradicting reports about the effect of caffeine on learning and memory. While some studies suggest an improving effect of caffeine administration in animal and human models, other reports that caffeine do not affect memory or even impairs. In the present study, we investigated whether caffeine administration alters late memory consolidation and fear memory persistence in contextual conditioning task in rats. Male adult Wistar rats received three 1 s - 0.6 mA footshocks (40 s apart) in a fear conditioning chamber and were injected with saline (0.9 % NaCl, i.p.) or caffeine (0.3, 3 or 30 mg/kg, i.p.) or spermidine (10 mg/kg, i.p.), 12 hours post-training. The testing session was held at 2, 7 or 14 days post-training and the percent of freezing responses was measured. Caffeine administration (3 mg/kg, i.p.) 12 h post-training increased freezing to context of rats tested 2 days after training. Other dose of caffeine were not able to alter freezing to context in the testing session at 7 and 14 days after training. Spermidine administration (10 mg/kg, i.p.) 12 h post-training was able to increase the freezing to context in the testing session carried 2 and 7 days after training. Our findings suggest that late caffeine administration facilitates memory consolidation but not memory persistence in rats. / Cafeína, um antagonista não seletivo dos receptores de adenosina, é uma das substâncias psicoestimulantes mais consumidas no mundo. Existem relatos contraditórios a respeito do efeito da cafeína sobre a memória e aprendizado. Enquanto alguns estudos sugerem um efeito de melhora pela administração da cafeína em modelos animais de laboratório e humanos, outros relatam que ela não afeta a memória ou mesmo prejudique. No presente estudo, nós investigamos se a administração de cafeína altera a consolidação tardia e a persistência da memória de medo na tarefa de condicionamento ao contexto em ratos. Ratos Wistar machos adultos receberam três choques 1 s - 0,6 mA (separados por 40 s) em uma câmara de condicionamento de medo, e foram injetados com salina (0,9 % NaCl, i.p.) ou cafeína (0,3, 3 ou 30 mg/kg, i.p.) ou espermidina (10 mg/kg, i.p.), 12 horas após o treino. A sessão de teste foi realizada 2, 7 ou 14 dias após o treino e a porcentagem da resposta de freezing foi medida. A administração de cafeína (3 mg/kg, i.p.) 12 h após o treino aumentou o freezing ao contexto de ratos testados 2 dias após o treino. Nenhuma dose de cafeína foi capaz de alterar o freezing ao contexto nas sessões de teste realizadas 7 e 14 dias após o treino. A administração de espermidina (10 mg/kg, i.p.) 12 h após o treino, foi capaz de aumentar o freezing ao contexto nas sessões de teste realizadas 2 e 7 dias após o treino. Nossos resultados sugerem que a administração tardia de cafeína facilita a consolidação da memória, mas não a persistência da memória em ratos.

Les réseaux vasculaires cérébraux adaptent leur activité à la demande métabolique des neurones environnants, mais leur contribution fonctionnelle à la consolidation de la mémoire, processus par lequel les traces mnésiques se stabilisent dans le temps, reste inconnue. A l’aide d’un test de mémoire olfactif associatif couplé à des approches biochimiques et d’imagerie cérébrale chez le rat, nous avons étudié la dynamique des changements vasculaires au cours de la consolidation mnésique qui nécessite une interaction transitoire entre l'hippocampe et les régions corticales constituant les sites dépositaires des souvenirs. Nous montrons que la formation d’une mémoire durable est associée, dès l’encodage, à un signal hypoxique qui déclenche une angiogenèse transitoire dans des régions corticales spécifiques impliquées plus tard dans le stockage des souvenirs. Manipuler cette angiogenèse corticale précoce (ACP) par blocage ou stimulation spécifique de la voie de signalisation de l'angiopoïétine-2 perturbe, ou améliore, le rappel des informations anciennement acquises. Stimuler l’ACP chez un modèle de rats hypertendus présentant des déficits d’activation de la voie de l’angiopoïetine-2 et de formation de la mémoire pallie le déficit mnésique observé, confirmant l'importance fonctionnelle de l’ACP comme un prérequis à la formation des souvenirs. L'hypergravité, connue pour altérer les fonctions vasculaires, n’a pas modifié l'organisation de la mémoire. Nos résultats identifient l’ACP comme un processus neurobiologique crucial sous-tendant la formation et la stabilisation des souvenirs. Ils révèlent l'importance de la plasticité vasculaire dans la modulation des fonctions cognitives et suggèrent que les changements structurels précoces du réseau vasculaire cérébral constituent un mécanisme permissif pour la régulation de la plasticité neuronale au sein des réseaux corticaux impliqués dans la formation progressive et le stockage des souvenirs. / While the cerebral microvasculature is known to adapt its activity according to the metabolic demand of surrounding neurons, the functional contribution of vascular networks to memory consolidation, the process by which memory traces acquire stability over time, remains elusive. By using an associative olfactory memory task in rats coupled to biochemical and imaging techniques, we investigated the dynamics of vascular changes during memory consolidation which requires a transitory interaction between the hippocampus and distributed cortical regions that ultimately support storage of enduring memories. We found that remote memory formation was associated, upon encoding, with a hypoxic signal that triggered transitory angiogenesis in specific cortical regions which support memory storage and retrieval only weeks later. Manipulating early cortical angiogenesis (ECA) by selectively blocking or stimulating the angiopoietin-2 signaling pathway impaired or improved remote memory retrieval, respectively. Enhancing ECA in spontaneously hypertensive rats, which exhibit reduced angiopoietin- 2 expression when cognitively challenged and are unable to properly stabilize and/or retrieve remotely acquired information, was efficient in rescuing the observed memory deficit, thus confirming the functional importance of ECA as a prerequisite for the formation of remote memories. Hypergravity, known to impair vascular functions, failed to alter the organization of recent and remote memory. Altogether, our findings identify ECA as a crucial neurobiological process underlying the formation and stabilization of remote memory. They highlight the importance of vascular plasticity in modulating cognitive functions and suggest that the early structural changes within vascular networks constitute a permissive mechanism for the regulation of neuronal plasticity within cortical networks which support the formation and storage of enduring memories.

Muitas evidências demonstram que o peptídeo liberador de gastrina (GRP) é um fator de crescimento que afeta funções neuroendócrinas, incluindo proliferação e diferenciação celular, comportamento alimentar, formação de memória, respostas a estresses, desenvolvimento de neoplasias, desordens neurológicas e psiquiátricas. Porém, os eventos moleculares pelos quais isso ocorre ainda não são totalmente compreendidos. No presente estudo, nós avaliamos as interações entre o receptor do peptídeo liberador de gastrina (GRPR) e a via de sinalização celular da PKA, tanto na proliferação celular de glioblastoma humano (in vitro) quanto na consolidação da memória no hipocampo de ratos Wistar (in vivo). Mostramos que o GRP age em sinergismo com agentes que estimulam a via do cAMP/PKA, promovendo a proliferação de células de glioblastoma humano, pois o tratamento com GRP combinado com um ativador de adenilil ciclase (AC), forskolin, ou um análogo de cAMP, 8-Br-cAMP, ou um inibidor do tipo IV de fosfodiesterase, rolipram, aumentaram a proliferação das células de U- 138MG, quando avaliadas pelo método de MTT. Nenhum destes compostos teve efeito sozinho. O mRNA de GRPR e a expressão protéica em U-138MG foram detectados pelas técnicas de RT-PCR e imuno-histoquímica. No estudo in vivo a bombesina em baixas doses induziu um aumento na consolidação da memória. O resultado foi potencializado na combinação com um ativador do receptor de dopamina D1/D5 (D1R), além de ser prevenido quando combinado com um inibidor da via da PKA. Os resultados sugerem que GRP e GRPR interagem com a via de sinalização cAMP/PKA tanto na estimulação da proliferação celular em linhagem de câncer humano quanto na modulação da memória no hipocampo de ratos. / Increasing evidence indicates that gastrin-releasing peptide (GRP) acts as an autocrine growth factor for brain tumors as well as been implicated in memory formation, however, underlying molecular events are poorly understood. In the present study, we examined interactions between the GRPR and cellular signaling pathways in influencing memory consolidation in the hippocampus and on proliferation of glioblastoma cell in vitro. We show here that GRP acts synergistically with agents that stimulate the cAMP/PKA pathway to promote proliferation of human gliobastoma cells. Treatment with GRP combined with the adenylyl cyclase (AC) activator forskolin, the cAMP analog 8-Br-cAMP, or the phosphodiesterase type IV (PDE4) inhibitor rolipram increased proliferation of U138-MG cells in vitro measured by MTT assay. None of the compounds had an effect when given alone. GRP receptor (GRPR) mRNA and protein expression in U138-MG cells was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. We investigated the interactions between the GRPR and the PKA pathway in male Wistar rats. BB-induced enhancement of consolidation was potentiated by co infusion of activators of the dopamine D1/D5 receptor (D1R) pathway and prevented by a PKA inhibitor. The results suggest that GRP and the GRPR interact with the cAMP/PKA signaling pathway in stimulating a cancer cell line proliferation and in memory modulation by hippocampal.

Muitas evidências demonstram que o peptídeo liberador de gastrina (GRP) é um fator de crescimento que afeta funções neuroendócrinas, incluindo proliferação e diferenciação celular, comportamento alimentar, formação de memória, respostas a estresses, desenvolvimento de neoplasias, desordens neurológicas e psiquiátricas. Porém, os eventos moleculares pelos quais isso ocorre ainda não são totalmente compreendidos. No presente estudo, nós avaliamos as interações entre o receptor do peptídeo liberador de gastrina (GRPR) e a via de sinalização celular da PKA, tanto na proliferação celular de glioblastoma humano (in vitro) quanto na consolidação da memória no hipocampo de ratos Wistar (in vivo). Mostramos que o GRP age em sinergismo com agentes que estimulam a via do cAMP/PKA, promovendo a proliferação de células de glioblastoma humano, pois o tratamento com GRP combinado com um ativador de adenilil ciclase (AC), forskolin, ou um análogo de cAMP, 8-Br-cAMP, ou um inibidor do tipo IV de fosfodiesterase, rolipram, aumentaram a proliferação das células de U- 138MG, quando avaliadas pelo método de MTT. Nenhum destes compostos teve efeito sozinho. O mRNA de GRPR e a expressão protéica em U-138MG foram detectados pelas técnicas de RT-PCR e imuno-histoquímica. No estudo in vivo a bombesina em baixas doses induziu um aumento na consolidação da memória. O resultado foi potencializado na combinação com um ativador do receptor de dopamina D1/D5 (D1R), além de ser prevenido quando combinado com um inibidor da via da PKA. Os resultados sugerem que GRP e GRPR interagem com a via de sinalização cAMP/PKA tanto na estimulação da proliferação celular em linhagem de câncer humano quanto na modulação da memória no hipocampo de ratos. / Increasing evidence indicates that gastrin-releasing peptide (GRP) acts as an autocrine growth factor for brain tumors as well as been implicated in memory formation, however, underlying molecular events are poorly understood. In the present study, we examined interactions between the GRPR and cellular signaling pathways in influencing memory consolidation in the hippocampus and on proliferation of glioblastoma cell in vitro. We show here that GRP acts synergistically with agents that stimulate the cAMP/PKA pathway to promote proliferation of human gliobastoma cells. Treatment with GRP combined with the adenylyl cyclase (AC) activator forskolin, the cAMP analog 8-Br-cAMP, or the phosphodiesterase type IV (PDE4) inhibitor rolipram increased proliferation of U138-MG cells in vitro measured by MTT assay. None of the compounds had an effect when given alone. GRP receptor (GRPR) mRNA and protein expression in U138-MG cells was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. We investigated the interactions between the GRPR and the PKA pathway in male Wistar rats. BB-induced enhancement of consolidation was potentiated by co infusion of activators of the dopamine D1/D5 receptor (D1R) pathway and prevented by a PKA inhibitor. The results suggest that GRP and the GRPR interact with the cAMP/PKA signaling pathway in stimulating a cancer cell line proliferation and in memory modulation by hippocampal.

Muitas evidências demonstram que o peptídeo liberador de gastrina (GRP) é um fator de crescimento que afeta funções neuroendócrinas, incluindo proliferação e diferenciação celular, comportamento alimentar, formação de memória, respostas a estresses, desenvolvimento de neoplasias, desordens neurológicas e psiquiátricas. Porém, os eventos moleculares pelos quais isso ocorre ainda não são totalmente compreendidos. No presente estudo, nós avaliamos as interações entre o receptor do peptídeo liberador de gastrina (GRPR) e a via de sinalização celular da PKA, tanto na proliferação celular de glioblastoma humano (in vitro) quanto na consolidação da memória no hipocampo de ratos Wistar (in vivo). Mostramos que o GRP age em sinergismo com agentes que estimulam a via do cAMP/PKA, promovendo a proliferação de células de glioblastoma humano, pois o tratamento com GRP combinado com um ativador de adenilil ciclase (AC), forskolin, ou um análogo de cAMP, 8-Br-cAMP, ou um inibidor do tipo IV de fosfodiesterase, rolipram, aumentaram a proliferação das células de U- 138MG, quando avaliadas pelo método de MTT. Nenhum destes compostos teve efeito sozinho. O mRNA de GRPR e a expressão protéica em U-138MG foram detectados pelas técnicas de RT-PCR e imuno-histoquímica. No estudo in vivo a bombesina em baixas doses induziu um aumento na consolidação da memória. O resultado foi potencializado na combinação com um ativador do receptor de dopamina D1/D5 (D1R), além de ser prevenido quando combinado com um inibidor da via da PKA. Os resultados sugerem que GRP e GRPR interagem com a via de sinalização cAMP/PKA tanto na estimulação da proliferação celular em linhagem de câncer humano quanto na modulação da memória no hipocampo de ratos. / Increasing evidence indicates that gastrin-releasing peptide (GRP) acts as an autocrine growth factor for brain tumors as well as been implicated in memory formation, however, underlying molecular events are poorly understood. In the present study, we examined interactions between the GRPR and cellular signaling pathways in influencing memory consolidation in the hippocampus and on proliferation of glioblastoma cell in vitro. We show here that GRP acts synergistically with agents that stimulate the cAMP/PKA pathway to promote proliferation of human gliobastoma cells. Treatment with GRP combined with the adenylyl cyclase (AC) activator forskolin, the cAMP analog 8-Br-cAMP, or the phosphodiesterase type IV (PDE4) inhibitor rolipram increased proliferation of U138-MG cells in vitro measured by MTT assay. None of the compounds had an effect when given alone. GRP receptor (GRPR) mRNA and protein expression in U138-MG cells was detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. We investigated the interactions between the GRPR and the PKA pathway in male Wistar rats. BB-induced enhancement of consolidation was potentiated by co infusion of activators of the dopamine D1/D5 receptor (D1R) pathway and prevented by a PKA inhibitor. The results suggest that GRP and the GRPR interact with the cAMP/PKA signaling pathway in stimulating a cancer cell line proliferation and in memory modulation by hippocampal.