O aumento da obesidade infantil tem sido correlacionado com a prevalência da puberdade precoce. Em animais pré-puberes verifica-se que a administração da dieta hiperlipídica induz a antecipação da puberdade. Estudos sugerem que a leptina pode influenciar o eixo reprodutivo por meio de sua ação em neurônios que expressam kisspeptina, no hipotálamo. Uma subpopulação de neurônios kisspeptidérgicos do núcleo arqueado coexpressa a dinorfina e neuroquinina B, que exercem uma ação autorreguladora sobre a secreção de kisspeptina. Considerando que a leptina, a kisspeptina, neuroquinina B e dinorfina são apontados como reguladores do início da puberdade e da função reprodutiva, este trabalho investigou como a dieta hiperlípidica (HFD) pode influenciar esses moduladores ao longo do início do desenvolvimento puberal e após a maturação sexual. Camundongos fêmeas Kiss1-hrGFP foram tratadas com a HFD ou dieta controle a partir do desmame e foram decapitadas aos 28, 32 e 63 dias (Diestro II) e o sangue coletado para dosagem plasmática de leptina, LH e estradiol. Um segundo grupo de fêmeas, tratadas com HFD ou dieta controle, receberam com 32 e 63 dias (Diestro I) uma injeção intraperitoneal de leptina (0,5 mg/kg) ou de veículo, 45 minutos após a injeção foram anestesiadas e submetidas à perfusão cerebral. Cortes hipotalâmicos foram utilizados para realização da técnica de imunoistoquímica para dupla marcação: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn e Kiss1-NKB. Verificamos que a HFD administrada após o desmame, quando comparada à dieta controle, antecipou a abertura vaginal, o primeiro estro e o ínicio da ciclicidade em fêmeas Kiss1-hrGFP. Como indicativo de ativação do eixo reprodutivo neste grupo, foi observado aumento do peso do útero e ovário, aos 32 dias. Esses efeitos estiveram associados com: a) o aumento da concentração plasmática de leptina; b) aumento da ativação de p-STAT3 no núcleo arqueado (ARC) e c) aumento de p-STAT3 em neurônios Kiss1-hrGFP e em neurônios CART do ARC em resposta ao estímulo da leptina. A HFD não afetou a colocalização entre Kiss1-Dyn e Kiss1-NKB em fêmeas no início do desenvolvimento puberal. Em fêmeas Kiss1-hrGFP (63 dias) a administração da HFD por 6 semanas não alterou a regularidade do ciclo estral, houve elevação da leptina plasmática associada com diminuição da expressão da p-STAT3 no ARC em resposta ao estímulo com leptina, sem alterar a responsividade dos neurônios Kiss1-hrGFP a este hormônio. Em conclusão, 9 os resultados demonstram que a antecipação da puberdade induzida pela HFD está associada com o aumento da ação central da leptina, que ao estimular o sistema kisspeptidérgico, direta ou indiretamente, por meio de neurônios CART, pode aumentar o tônus excitatório sobre o eixo reprodutivo. A administração da HFD após o desmame (6 semanas) promoveu uma redução à resposta da leptina no ARC na vida adulta, porém não afetou a responsividade dos neurônios Kiss1-hrGFP a este hormônio e nem a regularidade do ciclo estral nas fêmeas Kiss1-hrGFP. Em conjunto, esses resultados indicam que as ações da leptina nesses neurônios foram preservadas, mesmo com a resistência parcial de suas ações no ARC. Estudos adicionais deverão ser realizados para se avaliar os efeitos a longo prazo da HFD sobre o eixo reprodutivo. / The increase of childhood obesity has been correlated with the early onset of puberty. In prepubertal animals it appears high fat diet (HFD) induces anticipation of puberty. Several studies have indicated that the effect of leptin on reproductive axis can be mediated by its action on Kiss1 neurons in the hypothalamus.A subpopulation of Kiss1 neurons in the arcuate nucleus also shows co-expression of neurokinin B and dynorphin A,presenting a self-regulatory action kisspeptin secretion. Since leptin, kisspeptin, neurokinin B and dynorphin are identified as regulators of puberty onset and reproductive function, this study investigated the influence of high fat diet on these modulators along puberty onset and after sexual maturation. A group of Kiss1-hrGFP-female mice received the HFD or regular chow after weaning. The Kiss1-hrGFP females were decapitated at 28, 32 and 63 days old (Diestrus II) and blood was collected for measurement of plasma leptin, LH and estradiol. A second group of females, treated with HFD or chow after weaning, received at 32 and 63 days old (Diestrus I) an intraperitoneal injection of leptin (0.5 mg / kg) or vehicle, and 45 minutes after they were anesthetized and subjected to cerebral perfusion. Immunohistochemistry was performed for double labeling: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn and Kiss1-NKB in the hypothalamus. It was found that, compared to controls; the HFD anticipated the vaginal opening, the first estrus and the beginning of cyclicity in Kiss1-hrGFP females. An increase in the uterine weight and ovaries, confirmed the activation of reproductive axis in the HFD group at 32 days old. These effects were associated with: a) an increase in plasma leptin concentration; b) the activation of p-STAT3 in the arcuate nucleus (ARC) and c) the increase of p-STAT3 in Kiss1-hrGFP neurons and in CART neurons of the ARC in response to the stimulus of leptin. HFD didnt affect colocalization between Kiss1-Dyn and Kiss1-NKB in females mice at hte pubertal onset . Kiss1-hrGFP female mice (63 days old), exposed to HFD for 6 weeks,exhibited regular estrous cycles, increased plasma levels of leptin associated with a decreased p-STAT3 expression in the ARC after leptin stimulation, with no changes in the responsiveness of Kiss1-hrGFP neurons to this hormone, compared to controls. In conclusion, 11 the results demonstrate that the early puberty induced by the HFD is associated with increased central leptin action, by stimulating the kisspeptidergic system, directly or indirectly, through CART neurons that can increase the excitatory tone in the reproductive axis. The administration of HFD after weaning (6 weeks) induced a decrease on the response to leptin in the ARC in adulthood, but without changes in the responsiveness of Kiss1-hrGFP neurons to this hormone and the regularity of the estrous cycle in Kiss1-hrGFP females. Taken together, these results indicate that the actions of leptin in kisspeptin neurons were preserved after 6 weeks of HFD, despite the partial resistance to this hormone in the ARC. Additional studies should be conducted to evaluate the long-term effects of HFD in the reproductive axis.

Female reproductive senescence is widespread among mammalian species, but menopause is limited to species with menstrual cycles. While hormonal changes at menopause have profound impacts in the lives of women at middle age, the complex mechanisms underlying this process remain obscure. All three levels of the hypothalamic-pituitary-gonadal (HPG) axis are involved in reproductive aging, and evidence highlights a critical role for the dysregulation of gonadotropin-releasing hormone (GnRH) neurons, the hypothalamic cells that drive reproductive function. To investigate neuroendocrine mechanisms that may initiate and perpetuate reproductive decline at each step in the transition to acyclicity, I utilized an ovarian-intact middle-aged female rat model of natural reproductive senescence. These studies focused on three hypothalamic nuclei that are known to control GnRH activity: the anteroventral periventricular nucleus (AVPV), the site of positive hormone feedback onto GnRH neurons; the arcuate nucleus (ARC), the site of negative feedback; and the median eminence (ME), the site of GnRH release, with the following specific aims: 1) Characterize neuroendocrine gene and protein expression in female rats throughout the natural transition to acyclicity; 2) Determine the effects of chronic N-methyl-D-asparate receptor subunit 2b (NMDAR-NR2b) inhibition in acyclic females; and 3) Examine neuroendocrine gene expression during premature reproductive senescence after perturbation of the HPG axis. The results of these studies identified novel molecular and cellular changes with age and reproductive cycle status in the ARC and ME, two regions that are underappreciated for their roles in reproductive senescence. Surprisingly, few molecular targets were identified in the AVPV, a region that is much better-studied in this context. In the ME and ARC, I found changes in transcription factors and evidence of altered hormone feedback via changes in sex steroid hormone receptors and enzyme expression with reproductive aging. I also discovered decreased expression of genes for the excitatory neuropeptides, kisspeptin and neurokinin B, as well as decreased percentage of kisspeptin immunoreactive cells and their co-expression with estrogen receptor alpha in the ARC. And finally, in the ME, neurotrophic factor expression was changed with age, and the presence and phosphorylation state of the NR2b subunit of the NMDA receptor contributes to a greater inhibitory tone with acyclicity. Together these studies have identified novel pathways, especially in the ARC and ME, that are related to reproductive decline. Furthermore, changes in the hypothalamic neural and glial network of neurotransmitters, neuropeptides, hormone receptors and other transcription factors are likely contributing to altered responses to hormonal feedback and decreased excitatory drive for GnRH release. / text

Orientadores: Felix Guillermo Reyes Reyes, Claudio Antonio Barbosa de Toledo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-17T12:09:40Z (GMT). No. of bitstreams: 1 Freitas_ThaisFernandaPintodeAlmeida_M.pdf: 1160984 bytes, checksum: b9379e4fd16f451c0710b2ca37537085 (MD5) Previous issue date: 2011 / Resumo: Alimentar-se faz parte da cultura do ser humano nao estando unicamente associado a necessidades fisiologicas. Um alimento e constituido de diversas moleculas dentre elas os aminoacidos. O glutamato (GLU) e o anion de um dos principais aminoacidos encontrados nos alimentos que, alem de fazer parte da composicao dos alimentos, e uma molecula essencial para a fisiologia do ser humano. Pode tambem ser ingerido devido ao uso do aditivo alimentar glutamato monossodico (MSG). O GLU desempenha inumeras funcoes no organismo, dentre elas podemos citar: neurotransmissor excitatorio do sistema nervoso central, precursor de GABA e de aminoacidos como prolina e glutamina. Como neurotransmissor o GLU atua sobre quatro tipos de receptores: ionotropicos (AMPA, Kainato e NMDA) e metabotropicos (mGluR1-mGluR8) divididos em tres grupos de acordo com a sua homologia genetica e mecanismos de acao. Esses receptores estao presentes em praticamente todo o sistema nervoso central (SNC) e em outros orgaos como coracao, pulmao e intestino. O MSG e utilizado como realcador de sabor em todo o mundo, sendo que o descobrimento do gosto basico conferido pelo glutamato propiciou a producao industrial do seu sal, glutamato monossodico. O gosto basico conferido pelo glutamato e denominado de Umami, que traduzido do japones significa gosto bom, delicioso. Existem inumeros estudos sobre o uso do MSG como aditivo alimentar (funcao tecnologica), assim como sobre sua funcao fisiologica e seus efeitos no organismo de mamiferos. Organizacoes internacionais e agencias de regulamentacao de muitos paises tem reportado e/ou avaliado que o uso do MSG como aditivo alimentar e seguro. Todavia, alguns autores tem relatado efeitos adversos no sistema nervoso central (SNC) associados a exposicao ao MSG. Assim, o presente estudo teve como objetivo exibir dado morfologico sobre a localizacao do receptor mGluR8 no nucleo arqueado do hipotalamo (NARC) de ratos Wistar e avaliar o efeito da ingestao de dietas adicionadas de diferentes concentracoes de MSG (0% (controle), 1%, 2,5% e 5%) sobre o mGluR8. Tambem foi avaliado o ganho de peso corporeo entre os grupos de animais alimentados com as dietas adicionadas de diferentes concentracoes de MSG. Para evidenciar a presenca do receptor mGluR8 foi utilizada a tecnica de imunohistoquimica. Para avaliar o ganho de peso corporeo os animais foram pesados semanalmente. Todos os dados, tanto da contagem celular da tecnica de imunohistoquimica quanto da pesagem, foram analisados por analise de variancia. Os resultados obtidos indicam nao haver diferenca significativa (p <0,05) entre os ratos que ingeriram as dietas adicionadas das diferentes concentracoes de MSG, tanto para o ganho de peso corporeo como para a presenca de receptores mGluR8 no nucleo arqueado do hipotalamo (NARC) / Abstract: Food is part of human culture not only associated to physiological needs. Food is composed of several molecules among them amino acids. Glutamate (GLU) is the anion of one of the main amino acids found in foods that, besides being part of the food composition, is a molecule essential for human physiology. It can also be ingested due to the use of the food additive monosodium glutamate (MSG). The GLU performs many functions in the body, among them we could mention: excitatory neurotransmitter in the central nervous system, precursor of GABA and other amino acids such as proline and glutamine. As a neurotransmitter GLU acts on four types of receptors: ionotropic (AMPA, NMDA and kainate) and metabotropic (mGluR1-mGluR8) divided into three groups according to their genetic homology. These receptors are present in nearly all central nervous system (CNS) and other organs such as heart, lung and intestine. MSG is used as a flavor enhancer all over the world. The discovery of the basic taste due to glutamate, led to the industrial production of its salt, monosodium glutamate. The basic taste induced by glutamate is called Umami, which translated from Japanese, means good taste, delicious. There are numerous studies on the use of MSG as a food additive (technological function), as well as its physiological functions and its effects in the organism. International organizations and regulatory agencies of many countries have reported and / or evaluated that the use of MSG as a food additive is safe. However, some authors have reported adverse effects associated with exposure to MSG. Thus, this study aimed to assess the presences of the metabotropic receptor mGluR8 in the arcuate nucleus of the hypothalamus (ARH) of Wistar rats, and to evaluate the effects in the mGluR8 receptor resulting from the dietary intake of different concentrations of MSG (0% [control], 1%, 2 , 5% and 5%) during 90 days. Also, it was evaluated the body weight gain of the rats fed with the diets containing MSG in the different concentrations. To demonstrate the presence of the mGluR8 receptor immunohistochemistry technique was employed, and in order to elucidate the weight gain, the animals were weighed weekly. All the data, cell counts from the immunohistochemistry technique and from the rats weighing, were evaluated by analysis of variance. The results showed no significant difference (p<0.05) for both: body weight gain and the presence of mGluR8 receptors among the animals that were fed with the diets containing the different MSG levels / Mestrado / Engenharia de Alimentos / Mestre em Ciência de Alimentos

O aumento da obesidade infantil tem sido correlacionado com a prevalência da puberdade precoce. Em animais pré-puberes verifica-se que a administração da dieta hiperlipídica induz a antecipação da puberdade. Estudos sugerem que a leptina pode influenciar o eixo reprodutivo por meio de sua ação em neurônios que expressam kisspeptina, no hipotálamo. Uma subpopulação de neurônios kisspeptidérgicos do núcleo arqueado coexpressa a dinorfina e neuroquinina B, que exercem uma ação autorreguladora sobre a secreção de kisspeptina. Considerando que a leptina, a kisspeptina, neuroquinina B e dinorfina são apontados como reguladores do início da puberdade e da função reprodutiva, este trabalho investigou como a dieta hiperlípidica (HFD) pode influenciar esses moduladores ao longo do início do desenvolvimento puberal e após a maturação sexual. Camundongos fêmeas Kiss1-hrGFP foram tratadas com a HFD ou dieta controle a partir do desmame e foram decapitadas aos 28, 32 e 63 dias (Diestro II) e o sangue coletado para dosagem plasmática de leptina, LH e estradiol. Um segundo grupo de fêmeas, tratadas com HFD ou dieta controle, receberam com 32 e 63 dias (Diestro I) uma injeção intraperitoneal de leptina (0,5 mg/kg) ou de veículo, 45 minutos após a injeção foram anestesiadas e submetidas à perfusão cerebral. Cortes hipotalâmicos foram utilizados para realização da técnica de imunoistoquímica para dupla marcação: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn e Kiss1-NKB. Verificamos que a HFD administrada após o desmame, quando comparada à dieta controle, antecipou a abertura vaginal, o primeiro estro e o ínicio da ciclicidade em fêmeas Kiss1-hrGFP. Como indicativo de ativação do eixo reprodutivo neste grupo, foi observado aumento do peso do útero e ovário, aos 32 dias. Esses efeitos estiveram associados com: a) o aumento da concentração plasmática de leptina; b) aumento da ativação de p-STAT3 no núcleo arqueado (ARC) e c) aumento de p-STAT3 em neurônios Kiss1-hrGFP e em neurônios CART do ARC em resposta ao estímulo da leptina. A HFD não afetou a colocalização entre Kiss1-Dyn e Kiss1-NKB em fêmeas no início do desenvolvimento puberal. Em fêmeas Kiss1-hrGFP (63 dias) a administração da HFD por 6 semanas não alterou a regularidade do ciclo estral, houve elevação da leptina plasmática associada com diminuição da expressão da p-STAT3 no ARC em resposta ao estímulo com leptina, sem alterar a responsividade dos neurônios Kiss1-hrGFP a este hormônio. Em conclusão, 9 os resultados demonstram que a antecipação da puberdade induzida pela HFD está associada com o aumento da ação central da leptina, que ao estimular o sistema kisspeptidérgico, direta ou indiretamente, por meio de neurônios CART, pode aumentar o tônus excitatório sobre o eixo reprodutivo. A administração da HFD após o desmame (6 semanas) promoveu uma redução à resposta da leptina no ARC na vida adulta, porém não afetou a responsividade dos neurônios Kiss1-hrGFP a este hormônio e nem a regularidade do ciclo estral nas fêmeas Kiss1-hrGFP. Em conjunto, esses resultados indicam que as ações da leptina nesses neurônios foram preservadas, mesmo com a resistência parcial de suas ações no ARC. Estudos adicionais deverão ser realizados para se avaliar os efeitos a longo prazo da HFD sobre o eixo reprodutivo. / The increase of childhood obesity has been correlated with the early onset of puberty. In prepubertal animals it appears high fat diet (HFD) induces anticipation of puberty. Several studies have indicated that the effect of leptin on reproductive axis can be mediated by its action on Kiss1 neurons in the hypothalamus.A subpopulation of Kiss1 neurons in the arcuate nucleus also shows co-expression of neurokinin B and dynorphin A,presenting a self-regulatory action kisspeptin secretion. Since leptin, kisspeptin, neurokinin B and dynorphin are identified as regulators of puberty onset and reproductive function, this study investigated the influence of high fat diet on these modulators along puberty onset and after sexual maturation. A group of Kiss1-hrGFP-female mice received the HFD or regular chow after weaning. The Kiss1-hrGFP females were decapitated at 28, 32 and 63 days old (Diestrus II) and blood was collected for measurement of plasma leptin, LH and estradiol. A second group of females, treated with HFD or chow after weaning, received at 32 and 63 days old (Diestrus I) an intraperitoneal injection of leptin (0.5 mg / kg) or vehicle, and 45 minutes after they were anesthetized and subjected to cerebral perfusion. Immunohistochemistry was performed for double labeling: Kiss1-p-STAT3/ p-STAT3-CART/ Kiss1-Dyn and Kiss1-NKB in the hypothalamus. It was found that, compared to controls; the HFD anticipated the vaginal opening, the first estrus and the beginning of cyclicity in Kiss1-hrGFP females. An increase in the uterine weight and ovaries, confirmed the activation of reproductive axis in the HFD group at 32 days old. These effects were associated with: a) an increase in plasma leptin concentration; b) the activation of p-STAT3 in the arcuate nucleus (ARC) and c) the increase of p-STAT3 in Kiss1-hrGFP neurons and in CART neurons of the ARC in response to the stimulus of leptin. HFD didnt affect colocalization between Kiss1-Dyn and Kiss1-NKB in females mice at hte pubertal onset . Kiss1-hrGFP female mice (63 days old), exposed to HFD for 6 weeks,exhibited regular estrous cycles, increased plasma levels of leptin associated with a decreased p-STAT3 expression in the ARC after leptin stimulation, with no changes in the responsiveness of Kiss1-hrGFP neurons to this hormone, compared to controls. In conclusion, 11 the results demonstrate that the early puberty induced by the HFD is associated with increased central leptin action, by stimulating the kisspeptidergic system, directly or indirectly, through CART neurons that can increase the excitatory tone in the reproductive axis. The administration of HFD after weaning (6 weeks) induced a decrease on the response to leptin in the ARC in adulthood, but without changes in the responsiveness of Kiss1-hrGFP neurons to this hormone and the regularity of the estrous cycle in Kiss1-hrGFP females. Taken together, these results indicate that the actions of leptin in kisspeptin neurons were preserved after 6 weeks of HFD, despite the partial resistance to this hormone in the ARC. Additional studies should be conducted to evaluate the long-term effects of HFD in the reproductive axis.

Background: The neuropeptide S receptor (NPSR1) and its ligand neuropeptide S (NPS) have received increased attention in the last few years, as both establish a previously unknown system of neuromodulation. Animal research studies have suggested that NPS may be involved in arousal/wakefulness and may also have a crucial role in sleep regulation. The single nucleotide polymorphism (SNP) rs324981 in NPSR1 has begun to shed light on a function of the NPS-system in human sleep regulation. Due to an amino acid exchange, the T-allele leads to an increased sensitivity of the NPSR1. In the only genomewide association study to date on circadian sleep parameters in humans, an association was found between rs324981 and regular bedtime. However, the sleep parameters in this study were only measured by self-rating. Therefore, our study aimed to replicate these findings using an objective measure of sleep. Methods: The study included n = 393 white subjects (62–79 years) who participated in an actigraphic assessment for determining sleep duration, rest duration, sleep onset, rest onset and sleep onset latency. Genotyping of the SNP rs324981 was performed using the TaqMan OpenArray System. Results: The genotype at rs324981 was not significantly associated with rest onset (bedtime) or sleep onset (p = .146 and p = .199, respectively). However, the SNP showed a significant effect on sleep- and rest duration (p = .007 and p = .003, respectively). Subjects that were homozygous for the minor T-allele had a significantly decreased sleep- and rest duration compared to A-allele carriers. Conclusion: The results of this study indicate that the sleep pattern in humans is influenced by the NPS-system. However, the previously reported association between bedtime and rs324981 could not be confirmed. The current finding of decreased sleep duration in T/T allele carriers is in accordance with studies in rodents reporting similar results after NPS application.:Background; Methods; Results; Conclusions

Thesis (Ph.D.)--University of Florida, 2004. / Typescript. Title from title page of source document. Document formatted into pages; contains 142 pages. Includes Vita. Includes bibliographical references.

Le concept de programmation métabolique repose sur les études épidémiologiques pionnières menées par David Barker à la fin des années 1980 : une sous-nutrition maternelle durant la grossesse est un contributeur majeur au faible poids de naissance d’une part, et au développement du syndrome métabolique à l’âge adulte d’autre part. Ces dernières années, le concept de programmation métabolique a été étendu de l’état physiopathologique à l’état de santé. Le concept « Developmental Origins of Health and Disease » (DOHaD) postule que l’environnement périnatal (grossesse et allaitement) conditionne la physiologie de la descendance à long terme. Au delà d’une sous-nutrition périnatale, de nombreuses études expérimentales menées chez l’animal ont montré qu’un régime maternel hyperlipidique périnatal pouvait également entrainer des modifications métaboliques chez les descendants adultes.Au niveau central, le noyau arqué hypothalamique (ARC) est un régulateur important du métabolisme. L’environnement périnatal altère-t-il certaines régulations moléculaires dans l’ARC ? Chez les mammifères, l’expression du génome est modulée par de petits ARN d’une vingtaine de nucléotides, les microARN (miARN), inhibiteurs de la traduction des ARNm en protéines ou initiateurs de leur dégradation. Les miARN participent-ils à la régulation de l’expression génique dans l’ARC ? Un régime hyperlipidique modifie-t-il leur expression ? C’est l’hypothèse que j’ai testée au cours de ma thèse.Mon travail de thèse a consisté d’une part à caractériser physiologiquement un modèle de programmation métabolique par un régime maternel hyperlipidique chez le rat, et d’autre part à étudier les populations de miARN dans l’ARC de la descendance. Pour cela, des mères ont été nourries avec un régime équilibré ou différents régimes hyperlipidiques (HF1 et HF2) durant les périodes de gestation et lactation.Le régime maternel hyperlipidique HF1 entraine une hyperinsulinémie attestant une insulino-résistance qu’il soit donné durant la période périnatale ou à l’âge adulte. La conjonction d’un régime HF durant la période périnatale et à l’âge adulte entraine en plus une hyperglycémie.J’ai étudié la descendance de mères nourries avec le régime HF1 (mères HF) à cinq stades postnataux (P4, P8, P14, P21 et P28), un stade juvénile (P75) et un stade adulte (P216). J’ai également étudié l’éventuel effet additif d’un régime hyperlipidique donné à l’âge adulte.J’ai étudié les profils d’expression de plus de 500 miARN pour chaque ARC par la technologie de séquençage haut-débit. J’ai tout d’abord établi la cinétique d’expression des miARN de chacune de ces descendances. 10% à 20% des miARN montrent une différence d’expression supérieure à trois fois stade à stade. Ceci est vrai que les descendants soient nés de mères nourries avec le régime équilibré ou HF1. Le régime maternel HF1 montre peu d’impact sur les profils d’expressions de miARN à ces stades de développement précoces. A chaque stade, seuls 3% à 7% des miARN ont une expression statistiquement supérieure à trois fois. Ces données démontrent que les miARN présentent une expression robuste au cours du développement post-natal, c’est à dire durant la période de maturation des circuits neuronaux de l’ARC.Aucune différence n’est observée durant la période juvénile. Cependant à l’âge adulte, 8% des miARN ont une expression statistiquement supérieure à trois fois chez les descendants HF1. L’ensemble de ces résultats démontre que l’expression des miARN chez l’adulte, après une période de latence, reflète le contexte lipidique périnatal. L’expression des miARN de l’ARC, tout comme la sensibilité à l’insuline, est soumis à une programmation par le régime alimentaire maternel. / The concept of metabolic programming is based on the pioneering epidemiological studies realised by David Barker in the late 1980s : maternal undernutrition during pregnancy is a major contributor to low birth weight on one hand and the development of metabolic syndrome in adulthood on the other hand. In the last few years, the metabolic syndrom spreads to the“Developmental Origins of Health and Disease”(DOHaD) concept that postulates that the perinatal (pregnancy and lactation) environment influences the long term offspring’s physiology. Beyond the perinatal undernutrition impact, many experimental studies carried out on the animals have highlighted that a maternal perinatal high fat diet is responsible for metabolic disorders on the offspring.In the central nervous system, the arcuate nucleus (ARC) is an important regulator of metabolism. May an perinatal environment impares the molecular regulations in the ARC ? In the mammals, the genome expression is modulated by small RNAs of about twenty nucleotides, the microRNAs (miRNAs), that regulate, post-transcriptionnally, target mRNA degradation or translation. May the miARN participate to the gene expression regulation in the ARC ? May a high fat diet modify their expression ? This is the hypothesis tested in my thesis.My thesis work consisted on one hand to characterized a metabolic programming experimental model by a high fat maternal diet in the rat and on the other hand to study the miRNA populations of the ARC of the offspring. To this end, the dams were fed with a balanced or different high fat diet (HF1 and HF2) during gestation and lactation.The maternal HF1 diet lead to an hyperinsulinemia revealing the insulino-resistance when the diet is given during the perinatal periode or adulthood stages. The combination of a HF diet during the perinatal period and in adulthood leads to an hyperglycaemia.I have studied the offspring of dams fed HF1 (dams HF1) at five developmental stages (P4, P8, P14, P21 et P28), one juvenile stage (P75) and one adulthood stage (P216). I also studied the potential impact of an additionnal high fat diet given at adulthood stages.I have studied the expression pattern of over 500 miARN for each ARC by the high throughput sequencing technology. First of all, I have established the miARN expression kinetiks of each offspring. 10 percent to 20 percent of miARN have a fold-change more than 3 between stages. This is right for the offspring of HF dams or balanced dams. The maternal HF1 diet reveals few impacts on miRNA expression pattern during the early post-natal period. At each stages, only 3 percent to 7 percent of miARN have a fold-change more than 3. This results demonstating the high dynamics and robustness of miRNA populations in ARC at a time of maturation and circuitry organization of ARC neurons.No differences were observed during the juvenile period. However, in adulthood, 8% of miRNAs have a fold-change more than 3 in offspring HF1. All of these results demonstrating that the miARN expression of the adult, after a period of latence, reflects the perinatal lipidic context. The miARN expression of the ARC, as well as the insulin sensitivity, is submitted to a programming by a maternal diet.

L’intégrité fonctionnelle des organismes vivants, Homme y compris dépend des rythmes biologiques. La perturbation de ces rythmes due aux conditions de vie du monde moderne (travail posté, jet-lag,…) ou des circonstances naturelles (vieillissement), favorise l’installation de pathologies spécifiques (troubles du sommeil, l’obésité, le diabète,…). Afin de retarder l’apparition de ces troubles, la conception des projets expérimentaux sur des modèles d’animaux diurnes et des modèles d’animaux vivants dans des biotopes particuliers où l’Homme est présent (zone désertique) est nécessaire. Le but de ma thèse est d’essayer de comprendre les mécanismes neurophysiologiques d’adaptation aux conditions environnementales extrêmes chez le dromadaire (adapté à la chaleur) et le hamster d’Europe (adapté au froid). Une première partie révèle qu’en plus de la photopériode, la température ambiante est un véritable synchroniseur de l’horloge biologique du moins en absence du cycle lumière-obscurité. Dans une seconde partie, notre objectif était de vérifier si les variations saisonniers de la température ambiante sont capables comme les changements photopériodiques d’induire des modifications saisonnières sur le rythme de mélatonine et/ou de la température corporelle. Nous avons commencé par la vérification de l’existence de l’hétérothermie adaptative chez le dromadaire surtout que ce phénomène a été contesté récemment. Nos résultats démontrent que le dromadaire privé d’eau de boisson et placé sous des températures ambiantes élevées, adopte une régulation complexe de thermorégulation caractérisée par une alternance quotidienne de phase de poïkilothermie et d’homéothermie. Cet état d’hétérothermie a également été observé chez des dromadaires parfaitement hydratés, sujet d’une restriction alimentaire. Nos résultats concluent donc que l’hétérothermie adaptative chez le dromadaire est une combinaison de plusieurs facteurs qui interagissent le long du cycle lumière-obscurité, à savoir la température ambiante, la privation hydrique et la prise alimentaire. Finalement, nous avons démontré chez le hamster d’Europe que les neurones des noyaux arqués sont capables d’intégrer le signal photopériodique et cela indépendamment de la présence de la mélatonine. L’existence de ce mécanisme particulier d’intégration de la photopériode chez d’autres mammifères y compris l’Homme doit maintenant être recherchée. Nos résultats ouvrent la voie à la mise en évidence de son intérêt pratique dans le contrôle des rythmes biologiques en particulier dans celui des rythmes circannuels. / The functional integrity of living organisms, including human, depends on the biological rhythms. Disruption of these rhythms due to the living conditions of the modern world (shift work, jet lag ...) or natural circumstances (aging), leads various abnormalities (sleep disorders, obesity, diabetes ...). In order to understand pathophysiology of these abnormalities and adaptation in extreme environment, we need to design experiments on diurnal animals that cohabitate with human in specific biotopes. The aim of my thesis is to understand the neurophysiological mechanisms of adaptation to extreme environmental conditions in the camel (adapted to heat) and the European hamster (adapted to cold). Earlier we found that in addition to photoperiod, ambient temperature is a real synchronizer of the biological clock, at least in the absence of light-dark cycle. In the second part of my project, we investigated if seasonal variations of ambient temperature are capable to changes the rhythm of melatonin secretion and/or body temperature like photoperiod. Since heterothermy in camel challenged recently, we started our study by confirming the existence of adaptive heterothermy in camels. Our results demonstrate that dehydrated camels during exposure to daily heat show adaptive heterothermy. This mechanism is more complex because it is characterized by a daily alternation of two periods of poikilothermy and homeothermy. This adaptive heterothermy was also observed when camels are hydrated and food deprived. Based on our results, we can conclude that adaptive heterothermy in the Arabian camel is a combination of three factors interacting throughout the light-dark cycle: heat stress, water restriction, and level of food intake. Finally, we have demonstrated in European hamster that neurons of the arcuate nucleus are able to integrate photoperiodic signal, independent of melatonin. The existence of this particular mechanism of integration of photoperiod in other mammals including humans must be investigated. Our results promote to study the role of this new mechanism of integration of photoperiod in the control of biological rhythms in particular the circannual rhythms