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Le métabolisme énergétique cérébral via l’utilisation du glucose est fortement impliqué dans la production d’énergie nécessaire au fonctionnement du neurone en situation basale et d’activation. Des travaux précédents ont mis en évidence chez le rat chroniquement déficient en acides gras polyinsaturés (AGPI) de la série n-3 une altération de ce métabolisme en situation basale (diminution de l’utilisation cérébrale du glucose et de la densité des transporteurs de glucose GLUT1). Pour cerner les différentes étapes du métabolisme énergétique pouvant être modifiées par les AGPI n-3, l’expression des gènes clés a été mesurée par approche transcriptomique (cartes microfluidiques) chez l’animal déficient en AGPI n-3 ou supplémenté en acide docosahexaénoïque (DHA, 22 :6n-3). Ces mesures ont été réalisées sur deux zones cérébrales (cortex fronto-pariétal et couche CA1 de l’hippocampe) chez les animaux en situation basale et soumis à un environnement enrichi activant ces deux zones cérébrales. Pour ces 2 situations, le niveau d’utilisation cérébrale de glucose a été quantifié par la technique du fluoro-2-déoxyglucose couplée à l’imagerie de tomographie par émissions de positons (18FDG-TEP) chez les animaux déficients en AGPI n-3. L’analyse de la teneur cérébrale en AGPI membranaire a été réalisée par chromatographie en phase gazeuse et une approche in vitro sur culture primaire d’astrocytes a été développée pour apprécier l’impact du DHA sur les paramètres métaboliques de ces cellules.Les principaux résultats montrent que :- la déficience en n-3 diminue de 67% la teneur membranaire en DHA dans les deux zones cérébrales étudiées. Si la déficience induit principalement une diminution spécifique de l’expression de GLUT1 (-33%) dans le cortex fronto-pariétal en situation basale et d’activation, en revanche elle perturbe la neurotransmission glutamatergique dans l’hippocampe en augmentant l’expression des 2 transporteurs de glutamate (GLAST et GLT1). Par ailleurs, les données d’imagerie TEP mettent en évidence un hypométabolisme général du glucose chez les animaux déficients en n-3 en situation basale. Les données recueillies sur le modèle astrocytes soulignent un effet direct du DHA sur l’utilisation du glucose et l’expression de GLUT1 ;- La supplémentation en DHA ne modifie pas de façon appréciable la teneur membranaire en DHA dans les deux zones cérébrales étudiées. Au contraire de la déficience, il apparaît clairement pour la couche CA1 de l’hippocampe que l’expression de l’ensemble des gènes codant pour les complexes enzymatiques du cycle de krebs et de la voie de phosphorylation oxydative est significativement augmentée.Ces résultats originaux laissent ainsi entrevoir la possibilité que les acides gras de cette famille d’AGPI puissent intervenir sur l’énergétique et le fonctionnement de la synapse glutamatergique en modulant 1) le métabolisme glucidique (captage de glucose) et du glutamate en situation de déficit d’apport et 2) la production d’ATP (phosphorylation oxydative) en situation de supplémentation en DHA. L’altération de ces paramètres métaboliques au cours du vieillissement et dans certains désordres neurologiques, liée à un déficit de statut en DHA, mettent en avant les potentialités nutritionnelles des AGPI n-3 comme facteur préventif. / Cerebral energy metabolism via glucose utilization is heavily involved in the production of energy required to the neuron in basal conditions and activation. Previous work has shown in rats chronically deficient in n-3 polyunsaturated fatty acids (PUFA) altered the metabolism in basal condition (decrease of cerebral glucose use and density of glucose transporters GLUT1). To identify the different stages of energy metabolism may be modified by n-3 PUFA, the expression of key genes was measured by transcriptomic approach (Taqman Low Density Arrays) in animals deficient in n-3 PUFA or docosahexaenoic acid supplementation (DHA, 22:6n-3). These measurements were performed on two brain areas (fronto-parietal cortex and layer CA1 of the hippocampus) in animals in basal condition or submit to an enriched environment. For these two situations, the level of cerebral glucose utilization was quantified by the technique of fluoro-2-deoxyglucose imaging coupled with positron emission tomography (18FDG-PET) only in deficient n-3 PUFA animals. Analysis of brain PUFA content of membrane was performed by gas chromatography and an in vitro approach to primary culture of astrocytes was developed to assess the impact of DHA on metabolic parameters of these cells.The main results show that: - n-3 Deficiency decreases from 67% in membrane DHA content in both brain areas studied. If the deficiency induces mainly a decrease in the specific expression of GLUT1 (-33%) in the fronto-parietal cortex in basal and activation conditions, however it disrupts glutamatergic neurotransmission in the hippocampus by increasing the expression of two glutamate transporters (GLAST and GLT1). In addition, PET data show a general hypometabolism of glucose in animals deficient in n-3 in basal situation. Data collected on the model astrocytes point to a direct effect of DHA on glucose utilization and expression of GLUT1; - DHA supplementation does not alter significantly the membrane content of DHA in both brain areas studied. Unlike the n-3 deficiency, it is clear for the layer CA1 of the hippocampus that the expression of all genes encoding the enzyme complexes of the Krebs cycle and oxidative phosphorylation pathway is significantly increased. These original results suggest the possibility that the fatty acids of the n-3 PUFAs family can act on the energy and functioning of the glutamatergic synapse by modulating 1) glucose metabolism (glucose uptake) and glutamate in deficit intake situation and 2) the production of ATP (oxidative phosphorylation) in DHA supplementation. The alteration of these metabolic parameters during aging and certain neurological disorders, related to a deficit of DHA status, highlight the potential of dietary n-3 PUFA as a preventive factor.

The purpose of this study was to identify central neuronal sites activated by stimulation of cardiac ischemia-sensitive afferent neurons and determine whether electrical stimulation of left vagal afferent fibers modified the pattern of neuronal activation. Fos-like immunoreactivity (Fos-LI) was used as an index of neuronal activation in selected levels of cervical and thoracic spinal cord and brain stem. Adult Sprague-Dawley rats were anesthetized with urethane and underwent intrapericardial infusion of an "inflammatory exudate solution" (IES) containing algogenic substances that are released during ischemia (10 mM adenosine, bradykinin, prostaglandin E2, and 5-hydroxytryptamine) or occlusion of the left anterior descending coronary artery (CoAO) to activate cardiac ischemia-sensitive (nociceptive) afferent fibers. IES and CoAO increased Fos-LI above resting levels in dorsal horns in laminae I-V at C2 and T4 and in the caudal nucleus tractus solitarius. Dorsal rhizotomy virtually eliminated Fos-LI in the spinal cord as well as the brain stem. Neuromodulation of the ischemic signal by electrical stimulation of the central end of the left thoracic vagus excited neurons at the cervical and brain stem level but inhibited neurons at the thoracic spinal cord during IES or CoAO. These results suggest that stimulation of the left thoracic vagus excites descending inhibitory pathways. Inhibition at the thoracic spinal level that suppresses the ischemic (nociceptive) input signal may occur by a short-loop descending pathway via signals from cervical propriospinal circuits and/or a longer-loop descending pathway via signals from the nucleus tractus solitarius.

Serotonin (5-HT) has been implicated in energy homeostasis. There is growing evidence that 5-HT, acting through the 5-HT1BR in the paraventricular nucleus of the hypothalamus (PVN), is important to this regulation. To investigate the cellular events underlying 5-HT1BR action, a PVN neuronal cell model was established. The mHypoA-2/30 cell line expresses a complement of markers and neuropeptides specifically localized to the PVN. 5-HT induces neuronal activation in a dose-dependent manner as determined by an elevation in cFos mRNA levels. As 5-HT exerted limited transcriptional control, the integrity of 5-HT signaling machinery was assessed. 5-HT signals through cAMP and calcium secondary messenger systems by suppressing cAMP and elevating intracellular calcium, effects that are mimicked by activating the 5-HT1BR and that are attenuated in the presence of inhibitors. These findings support the use of this novel PVN cell model for delineating components involved in direct 5-HT action in PVN neurons.

Serotonin (5-HT) has been implicated in energy homeostasis. There is growing evidence that 5-HT, acting through the 5-HT1BR in the paraventricular nucleus of the hypothalamus (PVN), is important to this regulation. To investigate the cellular events underlying 5-HT1BR action, a PVN neuronal cell model was established. The mHypoA-2/30 cell line expresses a complement of markers and neuropeptides specifically localized to the PVN. 5-HT induces neuronal activation in a dose-dependent manner as determined by an elevation in cFos mRNA levels. As 5-HT exerted limited transcriptional control, the integrity of 5-HT signaling machinery was assessed. 5-HT signals through cAMP and calcium secondary messenger systems by suppressing cAMP and elevating intracellular calcium, effects that are mimicked by activating the 5-HT1BR and that are attenuated in the presence of inhibitors. These findings support the use of this novel PVN cell model for delineating components involved in direct 5-HT action in PVN neurons.

Submitted by Johnny Rodrigues (johnnyrodrigues@ufcg.edu.br) on 2018-04-04T21:48:36Z No. of bitstreams: 1 EUGÊNIO DE CARVALHO SARAIVA - DISSERTAÇÃO PPGCC 2014..pdf: 2813039 bytes, checksum: 9b76f48c8df4aee95923a8ce5f0385ce (MD5) / Made available in DSpace on 2018-04-04T21:48:36Z (GMT). No. of bitstreams: 1 EUGÊNIO DE CARVALHO SARAIVA - DISSERTAÇÃO PPGCC 2014..pdf: 2813039 bytes, checksum: 9b76f48c8df4aee95923a8ce5f0385ce (MD5) Previous issue date: 2014-07-30 / Há um número crescente de tecnologias que fazem uso de algoritmos de classificação para a automação de tarefas. Em particular, em Neurociências, algoritmos de classificação foram usados para testar hipóteses sobre o funcionamento do sistema nervoso central. No entanto, a relação entre as classes de padrões de ativação neuronal de áreas específicas do cérebro, como resultado de experiências sensoriais tem recebido pouca atenção. No contexto da Neurociência Computacional, este trabalho apresenta uma análise do nível de similaridade entre classes de padrões de ativação neuronal, com o uso das abordagens de aprendizagem não supervisionada e semi-supervisionada, em áreas específicas do cérebro de ratos em contato com objetos, obtidos durante um experimento envolvendo exploração livre de objetos pelos animais. As classes foram definidas de acordo com determinados tratamentos construídos com níveis específicos de um conjunto de 8 fatores (Animal, Região do Cérebro, Objeto ou Par de Objeto, Algoritmo de Agrupamento, Métrica, Bin, Janela e Intervalo de Contato). No total foram analisados 327.680 tratamentos. Foram definidas hipóteses quanto à relação de cada um dos fatores para com o nível de similaridade existente entre os tratamentos. As hipóteses foram verificadas por meio de testes estatísticos entre as distribuições que representavam cada uma das classes. Foram realizados testes de normalidade (Shapiro-Wilk, QQ-plot), análise de variância e um teste para diferenças entre tendência central (Kruskal-Wallis). Com base nos resultados encontrados nos estudos utilizando abordagem não supervisionada, foi inferido que os processos de aquisição e de definição dos padrões de ativação por um observador foram sujeitos a uma quantidade não significativa de ruídos causados por motivos não controláveis. Pela abordagem semisupervisionada, foi observado que nem todos os graus de similaridade entre pares de classes de objetos são iguais a um dado tratamento, o que indicou que a similaridade entre classes de padrões de ativação neuronal é sensível a todos os fatores analisados e fornece evidências da complexidade na codificação neuronal. / There are a growing number of technologies that make use of classification algorithms for automating tasks. In particular, in Neuroscience, classification algorithms were used to test hypotheses about the functioning of the central nervous system. However, the relationship between the classes of patterns of neuronal activation in specific brain areas as a result of sensorial experience has received little attention. In the context of Computational Neuroscience , this paper presents an analysis of the level of similarity between classes of patterns of neuronal activation with the use of learning approaches unsupervised and semi - supervised in specific areas of rat brain in contact with objects , obtained during an experiment involving free exploration of objects by animals. The classes were defined according to certain treatments constructed with specific levels with set of 8 factors (Animal, Brain Region, Object or Pair of Objects, Clustering Algorithm, Metric, Bin, Window and Interval Contact). In total 327.680 treatments were analyzed. Hypotheses regarding the relationship of each of the factors with the existing level of similarity between treatments were defined. The hypotheses were tested through between statistical distributions representing each class tests. The tests applied where the tests for normality (Shapiro-Wilk, QQ–plot), analysis of variance and a test for differences in central tendency (Kruskal-Wallis) were performed. Based on the results found in studies using an unsupervised approach, it was inferred that the process of acquisition and definition of patterns of activation by an observer was not subject to a significant amount of noise caused by uncontrollable reasons. For the semi-supervised approach, it was observed that not all degrees of similarity between pairs of classes of objects are equal to a given treatment, which indicated that the similarity between classes of patterns of neuronal activation is sensitive to all the factors analyzed and provides evidence about the complexity of neuronal coding.