Glucose and ketones are important energy substrates in the human body and brain. Their use is highly regulated depending on energy status which can vary according to multiple factors such as type of cell, fed or fasted state, type of diet, or health state. Use of either substrate is also subject to multiple homeostatic feedback loops. Energy substrate availability has implications in several disorders including declining cognitive function in the elderly. While glucose availability is known to decrease in elderly with cognitive deficits, it is unclear whether this also occurs in healthy elderly, either in the body or brain. Also unknown is whether, in healthy elderly, the use of ketones as energy substrates is affected, and whether ketones could be used as an alternative energy substrate in situations of a decline in glucose availability. A clearer understanding of the use of glucose and ketones in aging is necessary to determine whether declining energy substrate availability that may occur in the elderly is a contributing factor to cognitive deficits, a result of cognitive pathology, or simply a feature of the physiological aging process. Objective. The overall goal of the laboratory where this research was carried out is to ascertain whether alternate energy sources to glucose, i.e. ketones, may help alleviate the risk of declining cognitive function during aging. The specific objective of the research project presented in this thesis was to evaluate the metabolism of glucose and ketones in the healthy elderly compared to young or middle age subjects during mild, short-term ketosis induced by a ketogenic breakfast. Results. Elderly people in relatively good health have a similar capacity to produce ketones and to oxidize [superscript 13]C-glucose and [superscript 13]C-β-hydroxybutyrate as middle-aged or young adults. Discussion. The results of this project encourage further exploration of whether ketones could be used as and alternative energy substrate to glucose as, at least in healthy elderly, there is no impedance of raising plasma ketones in response to a ketogenic intervention. // Résumé : Introduction. Le glucose et les cétones sont des substrats énergétiques importants pour le corps et le cerveau humain. Leur utilisation est spécifiquement régulée selon I'état énergétique qui varie en fonction du type de cellule, de I'état nourrie ou à jeun, du type de diète, de I'état de la santé. L'utilisation est également régulée par des voies de rétrocontrôle homéostatique. La disponibilité des substrats énergétiques est impliquée dans plusieurs désordres, dont le déclin des fonctions cognitives, chez les personnes âgées où une diminution de la disponibilité du glucose est démontrée. Cependant, il n'est pas encore connu si cette diminution est présente chez les personnes âgées en bonne santé ; soit dans le corps ou le cerveau. La capacité d'utiliser les cétones comme substrats énergétiques chez les personnes âgées saines et la possibilité d'utiliser les cétones comme substrat énergétique alternatif dans le cas d'un déclin de la disponibilité de glucose sont inconnues. Une meilleure compréhension de I'utilisation du glucose et des cétones sera nécessaire pour clarifier si une diminution de la disponibilité des substrats énergétique contribue au déclin cognitif, se manifeste à la suite des pathologies cognitives, ou encore est simplement une caractéristique du processus physiologique du vieillissement. Objectif. L'objectif principal du laboratoire est de déterminer si les sources d'énergie alternatives au glucose, c'est-à-dire les cétones, pourraient ralentir le déclin cognitif chez les personnes âgées. L'objectif du projet de recherche de ce mémoire était d'évaluer le métabolisme du glucose et des cétones chez les sujets âgés, d'âge moyen, et jeune après la prise d'un déjeuner induisant une faible cétogénèse de courte durée. Résultats. Les personnes âgées en santé ont une capacité similaire au sujet d'âge moyen et jeune à produire des cétones et à oxyder le [indice supérieur 13]C-glucose et le [indice supérieur 13]C-β- hydroxybutyrate. Perspectives. Les résultats de ce projet incitent à continuer à explorer si les cétones pourraient être utilisés comme substrats énergétiques afin de contourner le problème d'un déclin de I'utilisation du glucose, car il n'y a aucun obstacle dans la production des cétones suite a une intervention cétonique chez des sujets âgées en bonne santé.

Climatério é a fase da vida da mulher em que ocorrem alterações hormonais que culminam em alterações metabólicas peculiares desse período, como por exemplo, diminuição do gasto energético e ganho ponderal. Com isso, os objetivos do presente trabalho foram mensurar o gasto energético e calcular a oxidação de carboidratos e lipídios, nos momentos basal e após uma sobrecarga lipídica, de mulheres pós-menopausadas com excesso de peso, estratificadas de acordo com o nível do estradiol plasmático (E2), bem como analisar peptídeos reguladores da fome e da saciedade nos mesmos momentos. Os grupos de mulheres foram: grupo 1- E2 39 pg/mL, 2- 40 E2 59 pg/mL, e 3- E2 60 pg/mL. As voluntárias receberam uma única refeição de 1100 kcal e 72% de lipídios, a qual caracterizou a sobrecarga lipídica. O gasto energético foi medido por calorimetria indireta por um período de cinco horas, sendo que as mensurações foram realizadas nos momentos 30, 60, 90, 150, 210 e 270 minutos. Os peptídeos reguladores da fome e da saciedade foram analisados no momento basal, após 30 e 270 minutos da sobrecarga lipídica. Os grupos foram comparados pelo teste não paramétrico Kruskal-Wallis, e para a comparação entre os momentos no mesmo grupo, foi utilizado o modelo de regressão linear com efeitos mistos, considerando p<0,05(*). Quarenta e quatro mulheres pós-menopausadas foram estudadas, com idade de 55±5 anos (média±desvio padrão), índice de massa corporal de 31±4 kg/m² e tempo de pós-menopausa de 8±7 anos. De uma forma geral, o gasto energético de repouso foi de 1337±194* kcal/dia, aumentando para 1476±295* kcal/dia nos primeiros 30´ pós-prandiais, e para 1513±318* kcal/dia ao final. Entre os grupos, não houve diferença estatística significativa. Em relação à oxidação de carboidratos, o valor basal foi de 151±95 mg/min, diminuindo para 146±95 mg/min nos primeiros 30´, e para 122±44 mg/min ao final (valores convertidos para mg/min para facilitar a leitura). O valor basal da oxidação lipídica foi de 37±30* mg/min, aumentando para 52±24* mg/min nos primeiros 30´ pós-prandiais, e para 63±21* mg/min ao final. Entre os grupos, não houve diferença estatística significativa quanto à oxidação de carboidratos, e quanto aos lipídios, houve diferença apenas entre os grupos 1 (maiores valores, p=0,03) e 2 ao final do experimento , e entre os grupos 1 e 3 (maiores valores, p=0,04) no momento 150´. Quanto aos peptídeos, o valor basal da leptina foi de 47±29 ng/mL, diminuindo para 44±27 ng/mL nos primeiros 30´ pós-prandiais, e para 43±26 ng/mL ao final. O valor basal da grelina foi de 753±522 pg/mL, diminuindo para 705±420 pg/mL nos primeiros 30´, e para 561±343 pg/mL ao final. O valor basal do peptídeo YY foi de 92±40* pg/mL, aumentando para 153±63* pg/mL nos primeiros 30´, e para 214±66* pg/mL ao final. Quanto à colecistocinina, o valor aos 30´ foi de 1,8±2,3 pg/mL, aumentando para 2,2±1,7 pg/mL ao final. Entre os grupos, houve diferença estatística significativa apenas da colecistocinina entre os grupos 2 e 3 (maiores valores, p=0,04) ao final. Sendo assim, houve um aumento significativo e gradual do gasto energético após a sobrecarga lipídica; predomínio da oxidação de carboidratos em relação à de lipídios, havendo uma diminuição em sua oxidação e um aumento da oxidação lipídica após a sobrecarga. Quanto aos grupos, não houve diferença estatística significativa em relação ao gasto energético, bem como à oxidação de carboidratos e lipídios na maior parte do tempo. Os peptídeos leptina e grelina diminuíram após a sobrecarga lipídica, enquanto que o peptídeo YY e a colecistocinina aumentaram, não havendo diferença estatística significativa entre os grupos na maior parte do tempo. Dessa forma, as variáveis estudadas se mostraram independentes do nível de estradiol plasmático. / Climaterium is the phase of women´s life when hormonal alterations occur and lead to peculiar metabolic responses such as a decrease in energy expenditure and weight gain. Thus, the aims of this study were to measure the energy expenditure, calculate carbohydrate and lipid oxidation, at resting and after a lipid overload, of postmenopausal women stratified according to their plasma estradiol value (E2) and evaluate hunger and satiety regulatory peptides in the same moment. Women´s groups were: group 1- E2 39 pg/mL, 2- 40 E2 59 pg/mL, and 3- E2 60 pg/mL. The volunteers received a 1100 kcal and 72% fat meal, which characterized the lipid overload. Energy expenditure was measured by indirect calorimetry for a period of five hours, and measurements were performed at 30, 60, 90, 150, 210 and 270 minutes. The hunger and satiety regulatory peptides were analyzed in the resting, 30 and 270 minutes after the lipid overload. The groups were compared using the nonparametric Kruskal-Wallis test. Linear regression model with mixed effects was used to compare the moments of the same group, considering p<0.05(*). Forty-four postmenopausal women were studied. Their age was 55±5 years (mean±standard deviation), body mass index was 31±4 kg/m² and they were in menopause for 8±7 years. In general, the resting energy expenditure was 1337±194* kcal/day, increasing to 1476±295* kcal/d in the first postprandial 30´, and to 1513±318* kcal/d in the end. There was no statistically significant difference between groups. Regarding carbohydrate oxidation, the baseline value was 151±95 mg/min, decreasing to 146±95 mg/min in the first 30´ and to 122±44 mg/min in the end (values were converted to mg/min for readability). The baseline value of lipid oxidation was 37±30* mg/min, increasing to 52±24* mg/min in the first 30´ and to 63±21* mg/min in the end. There was no statistically significant difference in carbohydrate oxidation between groups, and regarding lipids, the only statistically significant difference was between groups 1 (higher values, p=0,03) and 2 at the end of the experiment, and between groups 1 and 3 (higher values, p=0,04) at time 150´. In relation to the peptides, the baseline leptin was 47±29 ng/mL, decreasing to 44±27 ng/mL within the first postprandial 30´ and to 43±26 ng/mL in the end. Baseline ghrelin was 753±522 pg/mL, decreasing to 705±420 pg/mL in the first 30´ and to 561±343 pg/mL in the end. Baseline peptide YY was 92±40* pg/mL, increasing to 153±63* pg/mL in the first 30´ and to 214±66* pg/mL in the end. Referring to cholecystokinin, the value at 30´ was 1.8±2.3 pg/mL, increasing to 2.2±1.7 pg/mL in the end. Among groups, the only statistically significant difference was related to cholecystokinin between groups 2 and 3 (higher values, p=0,04) in the end. So, after the lipid overload, there was a gradual and significant increase on energy expenditure, predominance of carbohydrate oxidation in relation to the lipids with a decrease on its oxidation, and an increase on lipid oxidation. In regard to comparison between groups, there was no statistically significant difference in relation to energy expenditure, as well as to oxidation of carbohydrates and lipids in most of the time. The peptides leptin and ghrelin decreased after the lipid overload, while peptide YY and cholecystokinin increased, with no statistically significant difference between groups for most of the time. Thus, the variables were independent of plasma estradiol levels.

Climatério é a fase da vida da mulher em que ocorrem alterações hormonais que culminam em alterações metabólicas peculiares desse período, como por exemplo, diminuição do gasto energético e ganho ponderal. Com isso, os objetivos do presente trabalho foram mensurar o gasto energético e calcular a oxidação de carboidratos e lipídios, nos momentos basal e após uma sobrecarga lipídica, de mulheres pós-menopausadas com excesso de peso, estratificadas de acordo com o nível do estradiol plasmático (E2), bem como analisar peptídeos reguladores da fome e da saciedade nos mesmos momentos. Os grupos de mulheres foram: grupo 1- E2 39 pg/mL, 2- 40 E2 59 pg/mL, e 3- E2 60 pg/mL. As voluntárias receberam uma única refeição de 1100 kcal e 72% de lipídios, a qual caracterizou a sobrecarga lipídica. O gasto energético foi medido por calorimetria indireta por um período de cinco horas, sendo que as mensurações foram realizadas nos momentos 30, 60, 90, 150, 210 e 270 minutos. Os peptídeos reguladores da fome e da saciedade foram analisados no momento basal, após 30 e 270 minutos da sobrecarga lipídica. Os grupos foram comparados pelo teste não paramétrico Kruskal-Wallis, e para a comparação entre os momentos no mesmo grupo, foi utilizado o modelo de regressão linear com efeitos mistos, considerando p<0,05(*). Quarenta e quatro mulheres pós-menopausadas foram estudadas, com idade de 55±5 anos (média±desvio padrão), índice de massa corporal de 31±4 kg/m² e tempo de pós-menopausa de 8±7 anos. De uma forma geral, o gasto energético de repouso foi de 1337±194* kcal/dia, aumentando para 1476±295* kcal/dia nos primeiros 30´ pós-prandiais, e para 1513±318* kcal/dia ao final. Entre os grupos, não houve diferença estatística significativa. Em relação à oxidação de carboidratos, o valor basal foi de 151±95 mg/min, diminuindo para 146±95 mg/min nos primeiros 30´, e para 122±44 mg/min ao final (valores convertidos para mg/min para facilitar a leitura). O valor basal da oxidação lipídica foi de 37±30* mg/min, aumentando para 52±24* mg/min nos primeiros 30´ pós-prandiais, e para 63±21* mg/min ao final. Entre os grupos, não houve diferença estatística significativa quanto à oxidação de carboidratos, e quanto aos lipídios, houve diferença apenas entre os grupos 1 (maiores valores, p=0,03) e 2 ao final do experimento , e entre os grupos 1 e 3 (maiores valores, p=0,04) no momento 150´. Quanto aos peptídeos, o valor basal da leptina foi de 47±29 ng/mL, diminuindo para 44±27 ng/mL nos primeiros 30´ pós-prandiais, e para 43±26 ng/mL ao final. O valor basal da grelina foi de 753±522 pg/mL, diminuindo para 705±420 pg/mL nos primeiros 30´, e para 561±343 pg/mL ao final. O valor basal do peptídeo YY foi de 92±40* pg/mL, aumentando para 153±63* pg/mL nos primeiros 30´, e para 214±66* pg/mL ao final. Quanto à colecistocinina, o valor aos 30´ foi de 1,8±2,3 pg/mL, aumentando para 2,2±1,7 pg/mL ao final. Entre os grupos, houve diferença estatística significativa apenas da colecistocinina entre os grupos 2 e 3 (maiores valores, p=0,04) ao final. Sendo assim, houve um aumento significativo e gradual do gasto energético após a sobrecarga lipídica; predomínio da oxidação de carboidratos em relação à de lipídios, havendo uma diminuição em sua oxidação e um aumento da oxidação lipídica após a sobrecarga. Quanto aos grupos, não houve diferença estatística significativa em relação ao gasto energético, bem como à oxidação de carboidratos e lipídios na maior parte do tempo. Os peptídeos leptina e grelina diminuíram após a sobrecarga lipídica, enquanto que o peptídeo YY e a colecistocinina aumentaram, não havendo diferença estatística significativa entre os grupos na maior parte do tempo. Dessa forma, as variáveis estudadas se mostraram independentes do nível de estradiol plasmático. / Climaterium is the phase of women´s life when hormonal alterations occur and lead to peculiar metabolic responses such as a decrease in energy expenditure and weight gain. Thus, the aims of this study were to measure the energy expenditure, calculate carbohydrate and lipid oxidation, at resting and after a lipid overload, of postmenopausal women stratified according to their plasma estradiol value (E2) and evaluate hunger and satiety regulatory peptides in the same moment. Women´s groups were: group 1- E2 39 pg/mL, 2- 40 E2 59 pg/mL, and 3- E2 60 pg/mL. The volunteers received a 1100 kcal and 72% fat meal, which characterized the lipid overload. Energy expenditure was measured by indirect calorimetry for a period of five hours, and measurements were performed at 30, 60, 90, 150, 210 and 270 minutes. The hunger and satiety regulatory peptides were analyzed in the resting, 30 and 270 minutes after the lipid overload. The groups were compared using the nonparametric Kruskal-Wallis test. Linear regression model with mixed effects was used to compare the moments of the same group, considering p<0.05(*). Forty-four postmenopausal women were studied. Their age was 55±5 years (mean±standard deviation), body mass index was 31±4 kg/m² and they were in menopause for 8±7 years. In general, the resting energy expenditure was 1337±194* kcal/day, increasing to 1476±295* kcal/d in the first postprandial 30´, and to 1513±318* kcal/d in the end. There was no statistically significant difference between groups. Regarding carbohydrate oxidation, the baseline value was 151±95 mg/min, decreasing to 146±95 mg/min in the first 30´ and to 122±44 mg/min in the end (values were converted to mg/min for readability). The baseline value of lipid oxidation was 37±30* mg/min, increasing to 52±24* mg/min in the first 30´ and to 63±21* mg/min in the end. There was no statistically significant difference in carbohydrate oxidation between groups, and regarding lipids, the only statistically significant difference was between groups 1 (higher values, p=0,03) and 2 at the end of the experiment, and between groups 1 and 3 (higher values, p=0,04) at time 150´. In relation to the peptides, the baseline leptin was 47±29 ng/mL, decreasing to 44±27 ng/mL within the first postprandial 30´ and to 43±26 ng/mL in the end. Baseline ghrelin was 753±522 pg/mL, decreasing to 705±420 pg/mL in the first 30´ and to 561±343 pg/mL in the end. Baseline peptide YY was 92±40* pg/mL, increasing to 153±63* pg/mL in the first 30´ and to 214±66* pg/mL in the end. Referring to cholecystokinin, the value at 30´ was 1.8±2.3 pg/mL, increasing to 2.2±1.7 pg/mL in the end. Among groups, the only statistically significant difference was related to cholecystokinin between groups 2 and 3 (higher values, p=0,04) in the end. So, after the lipid overload, there was a gradual and significant increase on energy expenditure, predominance of carbohydrate oxidation in relation to the lipids with a decrease on its oxidation, and an increase on lipid oxidation. In regard to comparison between groups, there was no statistically significant difference in relation to energy expenditure, as well as to oxidation of carbohydrates and lipids in most of the time. The peptides leptin and ghrelin decreased after the lipid overload, while peptide YY and cholecystokinin increased, with no statistically significant difference between groups for most of the time. Thus, the variables were independent of plasma estradiol levels.

Les travaux entrepris au cours de cette thèse ont eu pour but d'étudier la modulation de la transmission synaptique inhibitrice au niveau des réseaux moteurs spinaux, à la fois au cours du développement et après lésion de la moelle épinière. Le nouveau-né présente des activités motrices spontanées qui jouent un rôle important dans la maturation des muscles et des réseaux de neurones de la moelle épinière. Dans une première étude, nous avons identifié l'un des mécanismes impliqués dans la genèse de ces activités chez le rat nouveau-né in vitro. Nous avons démontré que l'activation des récepteurs GABA&#7424; au niveau des terminales d'afférences primaires joue un rôle majeur dans le déclenchement et la propagation de ces activités spontanées. Dans une deuxième étude, nous avons testé la robustesse des dépolarisations de nature GABAergique enregistrées in vitro, c'est-à-dire leur dépendance vis-à-vis des paramètres du milieu de perfusion. Nous avons démontré que l'action dépolarisante des neurotransmetteurs GABA/glycine au niveau des motoneurones et celle du GABA au niveau des terminales d'afférences primaires ne sont pas dues à une fourniture énergétique insuffisante. La dernière étude a été consacrée à la modulation de la transmission synaptique inhibitrice après lésion de la moelle épinière. Nous avons montré que l'activation des récepteurs 5-HT2 (R5-HT2), particulièrement celle de l'isoforme 5-HT2&#7424;, renforce le poids synaptique inhibiteur via une hyperpolarisation du potentiel d'équilibre des ions chlorure (ECl) et une augmentation d'expression de KCC2 au niveau de la membrane des motoneurones. / The aim of this thesis was to explore the modulation of the inhibitory synaptic transmission within the spinal motor networks, both during development and after SCI. Spontaneous movements are an ubiquitous feature of fetal and infant behavior. They provide signals that are important for the development of muscles and the assembly of neuronal networks in the spinal cord. In a first study, we characterized one of the mechanisms underlying spontaneous motor behaviors in the in vitro spinal cord preparation isolated from neonatal rats. We demonstrated that the GABA is playing a key role in promoting spontaneous activity through primary afferent depolarizations which reach firing threshold. In the second part of my thesis, we tested the robustness of the in vitro GABAergic depolarizations and their dependence on the aCSF parameters. We demonstrated that during development the depolarizing actions of GABA/glycine on motoneurons and GABA on primary afferent terminals are not due to inadequate energy supply. In the last part of my thesis, we focused on the modulation of the inhibitory synaptic transmission following SCI. We demonstrated that activation of the 5-HT2 receptors, particularly the 5-HT2&#7424; subtype, strengthens inhibitory synaptic transmission to spinal motoneurons by hyperpolarizing the reversal potential of Cl- ions (ECl) and by increasing the cell-membrane expression of KCC2. This phenomenon reduces spasticity after SCI in rats. Upregulation of KCC2 function by targeting 5-HT2&#7424; receptors therefore opens new therapeutic strategies for the treatment of spasticity following SCI.