Yes / Metabolic syndrome is a global public health problem and predisposes individuals to obesity, diabetes and cardiovascular disease. Although the underlying mechanisms remain to be elucidated, accumulating evidence has uncovered a critical role of adipokines. Chemerin, encoded by the gene Rarres2, is a newly discovered adipokine involved in inflammation, adipogenesis, angiogenesis and energy metabolism. In humans, local and circulating levels of chemerin are positively correlated with body mass index and obesity-related biomarkers. In this review, we discuss both peripheral and central roles of chemerin in regulating body metabolism. In general, chemerin is upregulated in obese and diabetic animals. Previous studies by gain or loss of function show an association of chemerin with adipogenesis, glucose homeostasis, food intake and body weight. In the brain, the hypothalamus integrates peripheral afferent signals including adipokines to regulate appetite and energy homeostasis. Chemerin increases food intake in seasonal animals by acting on hypothalamic stem cells, the tanycytes. In peripheral tissues, chemerin increases cell expansion, inflammation and angiogenesis in adipose tissue, collectively resulting in adiposity. While chemerin signalling enhances insulin secretion from pancreatic islets, contradictory results have been reported on how chemerin links to obesity and insulin resistance. Given the association of chemerin with obesity comorbidities in humans, advances in translational research targeting chemerin are expected to mitigate metabolic disorders. Together, the exciting findings gathered in the last decade clearly indicate a crucial multifaceted role for chemerin in the regulation of energy balance, making it a promising candidate for urgently needed pharmacological treatment strategies for obesity.

Chez les hamsters, les jours longs activent la reproduction. Cette activation photopériodique (AP) impliquerait la mélatonine, l’hormone thyroïdienne (HT) et les RFamides hypothalamiques. Si les jours courts inhibiteurs sont maintenus au-delà de 20 semaines, une réactivation photoréfractaire (RP) de l’axe hypothalamo pituitaire gonadotrope (HPG) est déclenchée. Les mécanismes de cette RP sont inconnus. Notre analyse de la cinétique des changements moléculaires induits par l’AP et la RP montre dans les deux cas une inhibition précoce de l’expression de la Désiodinase 3 (Dio3), l’enzyme catabolisant les HT, dans les tanycytes. Associée à une inhibition tardive du transporteur MCT8 des HT, la diminution de Dio3 pourrait créer un pic d’HT dans l’hypothalamus. Dans les 2 activations, Kisspeptine et RFRP3 augmentent plusieurs semaines après l’inhibition de Dio3 et l’activation de l’HPG. Aussi, une inhibition d’RFRP3 lors de l’AP n’affecte pas l’HPG, mettant en cause le rôle du RFRP3. Nous avons donc découvert une inhibition précoce de Dio3 pouvant induire l’activation saisonnière de l’HPG. Le régulateur saisonnier précoce de Dio3 reste à découvrir. / In hamsters, reproduction is activated by long days. This photoactivation (PA) supposedly involves melatonin, hypothalamic thyroid hormones (TH) and RFamide peptides. Maintaining inhibitory short days for over 20 weeks triggers a photorefractory reactivation of the hypothalamo pituito gonadotropic axis (HPG) The mechanisms of this PR are so far unknown. Our cinetic analysis of the dynamic molecular changes in PA and PR revealed a conserved early inhibition of tanycytic deiodinase 3 (Dio3), which catabolizes TH, in both activation mechanisms. Associated with a late decrease of the TH transporter MCT8, the inhibition of Dio3 could generate an early peak of hypothalamic TH. In both activations, RFamide upregulation occurs several weeks after the initial Dio3 inhibition. Also, pharmacological inhibition of RFRP3 during PA does not influence the HPG activity, questioning the role of RFRP3 in HPG activation. We have thus uncovered a so far unreported early Dio3 inhibition that could be sufficient to seasonally reactivate the gonadotropic axis. The seasonal regulator of Dio3 remains to be discovered.

Pas de résumé / Production of new neurons in the brain decreases dramatically with age due to progressive physiological depletion of stem and progenitor cell populations (NSCs). Recent studies indicate that circulating factors constitute a systemic aging milieu regulating the birth of new cells. Interestingly, some long-lived mouse strains such as Ames dwarf mutants, with low circulating levels of GH and IGF-1, show increased neurogenesis and preserved hematopoietic stem cell pool. Thus, the possibility that genes regulating lifespan and aging also quantitatively modulate stem cells in mammals is more and more explored. IGF-1 plays a pivotal role in aging in different species, and I am asking whether some of the well-known longevity effects resulting from down-regulation of this signaling pathway could be explained by local regulation of stem and progenitor cell compartments. To validate this hypothesis, I pursued a dual approach based on biological experiments and mathematical modeling. Using a novel triple transgenic mouse model, I inactivated IGF-1 signaling specifically in adult NSCs, and traced knockout cell lineages with a fluorescent reporter transgene. By analyzing the phenotype at different time points after KO induction, I could distinguish between short and long-term effects of IGF signaling on cellular regeneration and identify cumulative physiological consequences of down-regulation of this pathway using behavioral tests. In my mathematical models, the dynamics of regenerative cell populations were described by a set of differential equations depending on circulating “growth-factor like molecules” (GFs). My results suggest that in aging tissues, the optimal distribution of GFs is a function that decreases with time. In the olfactory system, I showed that inactivation of IGF signaling in adult NSCs enhanced long-term maintenance of neuroblasts and increased the overall production of neurons. Mutants started with the same number of adult-born neurons as controls one month after KO induction at 4 months of age, but ended up having significantly more differentiated cells integrating the olfactory bulb at long-term, i.e. at 16 months of age. This highly increased neurogenic activity occurred without depletion of neural stem/progenitor cell compartments. In contrast, IGF-1R deletion in adult hippocampal stem cells did not change neurogenesis dynamics, pointing out a niche-dependent effect of IGFs. The important cellular changes in the olfactory bulb led to improved olfactory memory and odor discrimination in aged mutants. Strikingly, mutants also displayed altered energy homeostasis and increased sensitivity to metabolic hormones, namely leptin and insulin. This metabolic shift could be linked to enhanced olfactory function, and to changes in hypothalamic neurogenesis. Indeed, we observed that IGF-1R deletion in hypothalamic stem cells (HySC) protected α-tanycyte pool from age-related decline and increased the number of newborn neurons in the hypothalamus. Taken together, my results validate the hypothesis that life-long inhibition of IGF signaling in adult NSCs delays age-related decline of neurogenesis, in a niche-dependent manner. These data also show that local modulation of neural cell replacement has important physiological effects at the level of the whole organism, pointing out a novel pathophysiological role for adult neurogenesis.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O estado nutricional perinatal tem influência persistente sobre o desenvolvimento neural e a função cognitiva. Em humanos e outros animais, a malnutrição protéica durante o período perinatal acarreta alterações permanentes, incluindo a síndrome metabólica na idade adulta. A alimentação é modulada principalmente por fatores neuronais e hormonais que chegam ao hipotálamo. As reservas de glicogênio hipotalâmicos são uma fonte de glicose em altas demandas energéticas, como durante o desenvolvimento dos circuitos neurais. Como alguns circuitos hipotalâmicos estão sendo formados durante o período de lactação, focamos o estudo nos efeitos da desnutrição protéica, durante os primeiros 10 dias de lactação, sobre as reservas de glicogênio em núcleos hipotalâmicos envolvidos no controle do metabolismo energético. Ratas grávidas foram aleatoriamente separadas individualmente em gaiolas e alimentadas ad libitum com uma dieta normoproteíca (22% proteína). Após o parto, cada mãe ficava com 6 filhotes machos. Durante os 10 primeiros o grupo experimental recebia uma dieta isenta de proteína (D) e o grupo controle uma dieta normoproteíca (C). Em P10 a marcação para as reservas de glicogênio era muito intensa nos animais C no núcleo arqueado (ARC) e eminência média (EM). Em P20 no animais C, as reservas eram menores em comparação com P10. Os animais D apresentaram uma marcação menor do que os controles. Após P45 foi difícil determinar diferença entre os grupos porque o as reservas estavam diminuídas. Nós também mostramos que os tanicitos eram as células que apresentavam reservas de glicogênio. Nossos dados reforçam que o estado nutricional materno durante a lactação é crítico para a maturação cerebral, pois a malnutrição materna resulta em menor marcação nas reservas de glicogênio no hipotálamo, o que pode ser crítico para o estabelecimento da circuitaria neural. / Perinatal nutrition has persistent influences on neural development and cognition. In humans and other animals protein malnutrition during the perinatal period causes permanent changes, inducing to adulthood metabolic syndrome. Feeding is mainly modulated by neural and hormonal inputs to the hypothalamus. Hypothalamic glycogen stores are a source of glucose in high energetic demands, as during development of neural circuits. As some hypothalamic circuits are formed during lactation, we attempt to study the effects of malnutrition, during the first 10 days of lactation, on glycogen stores in hypothalamic nuclei involved in the control of energy metabolism. Female pregnant rats were fed ad libitum with a normorprotein diet (22% protein). After delivery each dam was kept with 6 male pups. During the first 10 days of lactation dams from experimental group received a protein free diet and the control group a normoprotein diet. By P10 glycogen stores were very high in the arcuate nucleus and median eminence of control group. Glycogen stores decreased during development. In P20 control animals, glycogen stores were lower when compared to P10 control animals. Animals submitted to malnutrition presented a staining even lower than control ones. After P45 it was difficult to determine differences between control and diet groups because glycogen stores were reduced. We also showed that tanycytes were the cells presenting glycogen stores. Our data reinforce that maternal nutritional state during lactation may be critical for neurodevelopment since it resulted in a low hypothalamic glycogen store, which may be critical for establishment of neuronal circuitry.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O estado nutricional perinatal tem influência persistente sobre o desenvolvimento neural e a função cognitiva. Em humanos e outros animais, a malnutrição protéica durante o período perinatal acarreta alterações permanentes, incluindo a síndrome metabólica na idade adulta. A alimentação é modulada principalmente por fatores neuronais e hormonais que chegam ao hipotálamo. As reservas de glicogênio hipotalâmicos são uma fonte de glicose em altas demandas energéticas, como durante o desenvolvimento dos circuitos neurais. Como alguns circuitos hipotalâmicos estão sendo formados durante o período de lactação, focamos o estudo nos efeitos da desnutrição protéica, durante os primeiros 10 dias de lactação, sobre as reservas de glicogênio em núcleos hipotalâmicos envolvidos no controle do metabolismo energético. Ratas grávidas foram aleatoriamente separadas individualmente em gaiolas e alimentadas ad libitum com uma dieta normoproteíca (22% proteína). Após o parto, cada mãe ficava com 6 filhotes machos. Durante os 10 primeiros o grupo experimental recebia uma dieta isenta de proteína (D) e o grupo controle uma dieta normoproteíca (C). Em P10 a marcação para as reservas de glicogênio era muito intensa nos animais C no núcleo arqueado (ARC) e eminência média (EM). Em P20 no animais C, as reservas eram menores em comparação com P10. Os animais D apresentaram uma marcação menor do que os controles. Após P45 foi difícil determinar diferença entre os grupos porque o as reservas estavam diminuídas. Nós também mostramos que os tanicitos eram as células que apresentavam reservas de glicogênio. Nossos dados reforçam que o estado nutricional materno durante a lactação é crítico para a maturação cerebral, pois a malnutrição materna resulta em menor marcação nas reservas de glicogênio no hipotálamo, o que pode ser crítico para o estabelecimento da circuitaria neural. / Perinatal nutrition has persistent influences on neural development and cognition. In humans and other animals protein malnutrition during the perinatal period causes permanent changes, inducing to adulthood metabolic syndrome. Feeding is mainly modulated by neural and hormonal inputs to the hypothalamus. Hypothalamic glycogen stores are a source of glucose in high energetic demands, as during development of neural circuits. As some hypothalamic circuits are formed during lactation, we attempt to study the effects of malnutrition, during the first 10 days of lactation, on glycogen stores in hypothalamic nuclei involved in the control of energy metabolism. Female pregnant rats were fed ad libitum with a normorprotein diet (22% protein). After delivery each dam was kept with 6 male pups. During the first 10 days of lactation dams from experimental group received a protein free diet and the control group a normoprotein diet. By P10 glycogen stores were very high in the arcuate nucleus and median eminence of control group. Glycogen stores decreased during development. In P20 control animals, glycogen stores were lower when compared to P10 control animals. Animals submitted to malnutrition presented a staining even lower than control ones. After P45 it was difficult to determine differences between control and diet groups because glycogen stores were reduced. We also showed that tanycytes were the cells presenting glycogen stores. Our data reinforce that maternal nutritional state during lactation may be critical for neurodevelopment since it resulted in a low hypothalamic glycogen store, which may be critical for establishment of neuronal circuitry.

Yes / Long-term and reversible changes in body weight are typical of seasonal animals. Thyroid hormone (TH) and retinoic acid (RA) within the tanycytes and ependymal cells of the hypothalamus have been implicated in the photoperiodic response. We investigated signalling downstream of RA and how this links to the control of body weight and food intake in photoperiodic F344 rats. Chemerin, an inflammatory chemokine, with a known role in energy metabolism, was identified as a target of RA. Gene expression of chemerin (Rarres2) and its receptors were localised within the tanycytes and ependymal cells, with higher expression under long (LD) versus short (SD) photoperiod, pointing to a physiological role. The SD to LD transition (increased food intake) was mimicked by 2 weeks of ICV infusion of chemerin into rats. Chemerin also increased expression of the cytoskeletal protein vimentin, implicating hypothalamic remodelling in this response. By contrast, acute ICV bolus injection of chemerin on a 12h:12h photoperiod inhibited food intake and decreased body weight with associated changes in hypothalamic neuropeptides involved in growth and feeding after 24hr. We describe the hypothalamic ventricular zone as a key site of neuroendocrine regulation, where the inflammatory signal, chemerin, links TH and RA signaling to hypothalamic remodeling. / BBSRC (grant number BB/K001043/1) and the Scottish Government.

L’hypothalamus médiobasal contient de nombreux noyaux régulant l’homéostasie énergétique en réponse aux variations des signaux métaboliques périphériques, telles que les nutriments et les hormones, l’informant de l’état énergétique de l’individu. Parmi ces noyaux, le noyau arqué hypothalamique (NA) est considéré comme le noyau clé de la régulation de la prise alimentaire. En effet, il est capable de recevoir et d’intégrer les informations métaboliques périphériques, pour ensuite les relayer vers les autres noyaux hypothalamiques régulant la prise alimentaire. Dans ce contexte, l’accès des molécules périphériques au NA est une étape importante dans la régulation de la prise alimentaire. L’organisation des interfaces sang/cerveau à ce niveau est d’ailleurs très particulière, suggérant une régulation spécifique de l’accès des molécules périphériques vers le NA. En effet, deux types de vaisseaux sont retrouvés dans cette région cérébrale : 1- les vaisseaux de la barrière hématoencéphalique (BHE) dans le NA et 2- les vaisseaux fenêtrés dans l’éminence médiane (EM), un organe circumventriculaire (OCV) adjacent au NA. Alors que les vaisseaux de la BHE présentent des propriétés de barrière et régulent les échanges sang/NA, les vaisseaux de l’EM possèdent de nombreuses fenestrations facilitant les échanges sang/EM. Ces deux types de vaisseaux ont la particularité d’être contactés par des cellules épendymaires hautement spécialisées formant le bas du 3ème ventricule. Ces cellules, appelées « tanycytes », expriment des protéines de jonctions serrées suggérant leur participation à la régulation des échanges sang/cerveau dans cette région cérébrale. En effet, des études menées au sein du laboratoire ont montré que les tanycytes de l’EM, contactant les vaisseaux fenêtrés, expriment des protéines de jonctions serrées (JS) organisées en ceinture continue autour de leur pôle apical. Ces JS créent ainsi un épendyme étanche qui limite les échanges EM/LCR. A l’inverse, les tanycytes du NA, contactant les vaisseaux de la BHE, expriment des protéines de JS non organisées en leur pôle apical. L’épendyme du NA est ainsi perméable et favorise les échanges LCR/NA. Le but de mon travail de thèse a donc été de comprendre, en prenant en compte tous ces éléments -c’est-à-dire la présence de vaisseaux fenêtrés, de vaisseaux de la BHE et des tanycytes -, comment est organisé l’accès des signaux métaboliques périphériques vers le NA et si cet accès pouvait être modulé afin de contrôler l’homéostasie énergétique. Nos expériences ont montré que, chez la souris mâle adulte, une glucopénie induite par le jeûne ou par le 2-desoxyglucose induisait une réorganisation structurale des vaisseaux et de l’épendyme au niveau de l’EM et du NA, modifiant ainsi les échanges sang/cerveau. En effet, chez ces souris, nous avons observé une augmentation du nombre de vaisseaux fenêtrés au niveau de l’EM et du NA, ainsi qu’une réorganisation fonctionnelle des protéines de JS au niveau du ventricule : les tanycytes du NA, contactant des vaisseaux fenêtrés à présent, réorganisent leurs protéines de jonctions serrées (JS) afin d’assurer l’homéostasie cérébrale. Ces réorganisations induisent alors un meilleur accès des molécules périphériques vers le NA. De plus, nos résultats ont montré que cette plasticité est induite par le VEGF-A, produit localement par les tanycytes. En effet, la neutralisation du VEGF-A bloque la plasticité de l’EM/NA induite par l’hypoglycémie et perturbent la réponse physiologique hyperphagique lors de la réalimentation. Enfin, nos données supplémentaires indiquent que cette plasticité de l’EM/NA est conservée dans différents modèles alimentaires et se produit également au cours de la journée, suggérant son implication dans le contrôle circadien du comportement alimentaire. / The mediobasal hypothalamus contains numerous nuclei regulating energy homeostasis in response to peripheral metabolic signals. Among these nuclei, the arcuate nucleus of the hypothalamus (ARH) is considered to be a critical component of the neural circuits regulating energy balance. Indeed, the ARH is able to integrate the metabolic information carried by nutrients and peripheral hormones, and to transmit their message to secondary nuclei regulating food intake. Therefore, the delivery of peripheral molecules conveying metabolic information to the ARH is a critical step in the regulation of food intake. At the level of the ARH, the organization of blood-brain interface is indeed peculiar. Both the vessels of the blood brain barrier (BBB) and the fenestrated vessels of the median eminence (ME), a circumventricular organ adjacent to the ARH, represent two pathways by which peripheral molecules may reach the ARH. While BBB vessels possess barrier properties restricting the access of peripheral molecules to the ARH, the vessels in the ME possess numerous fenestrations facilitating blood-brain exchanges. These two types of vessels are contacted by specialized ependymal cells lining the floor of the 3rd ventricle called tanycytes. Tanycytes express tight junction (TJ) proteins providing a putative role in the regulation of blood/brain exchanges. While ME tanycytes contact the fenestrated vessels which lie in the ME, they express TJ proteins organized as a continuous belt around their apical pole, creating a tight ependyma limiting ME/cerebrospinal fluid exchanges. In contrast, ARH tanycytes contact BBB vessels and express TJ proteins in a disorganized pattern thus creating a permeable ependyma allowing CSF/ARH exchanges. In the following studies we wished to examine how peripheral signals may reach the ARH through this peculiar blood-brain interface. Moreover, we wished to determine whether the blood-brain interface undergoes dynamic remodeling according to the energetic status of individual. To these aims the plasticity of the BBB vessels, fenestrated vessels of the ME, and tanycytes were analyzed under various metabolic challenges. Our studies show that both fasting- or 2-deoxyglucose-induced glucopenia induce vascular and ependymal reorganization in the ME and the ARH. This reorganization is characterized by an increase in the number of fenestrated vessels in the ME and in the ARH, and a reorganization of TJ proteins in ARH tanycytes, and led to improved access of peripheral molecules to the ARH. Furthermore, our results reveal that VEGF-A expression in tanycytes modulates the plasticity at the blood/brain interface. Indeed, the neutralization of VEGF signaling blocks fasting-induced barrier remodeling and significantly impairs the physiological response to refeeding. Finally, our supplementary results show that the ME-ARH reorganization is also observed in mice under different diet, as well as implicated in the circadian regulation of food intake. Strikingly, the ME-ARH organization is disturbed in diet-induced obese mice, which could be the origin of hormonal resistances observed in these mice. Altogether, our results suggest a new concept in the regulation of the food intake: peripheral glucose modulates blood/brain interface in the ME through a VEGF-dependent mechanism to improve the access of the metabolic signals towards the ARH. As other circumventricular organs possess a similar organization, characterized by fenestrated vessels and a tanycyte barrier, these exciting results pave the way to for future studies examining the remodeling of the blood-brain interface and its role in other neuroendocrine processes.

Yes / Seasonal rhythms in energy balance are well documented across temperate and equatorial zones animals. The long-term regulated changes in seasonal physiology consists of a rheostatic system that is essential to successful time annual cycles in reproduction, hibernation, torpor, and migration. Most animals use the annual change in photoperiod as a reliable and robust environmental cue to entrain endogenous (i.e. circannual) rhythms. Research over the past few decades has predominantly examined the role of first order neuroendocrine peptides for the rheostatic changes in energy balance. These anorexigenic and orexigenic neuropeptides in the arcuate nucleus include neuropeptide y (Npy), agouti-related peptide (Agrp), cocaine and amphetamine related transcript (Cart) and pro-opiomelanocortin (Pomc). Recent studies also indicate that VGF nerve growth factor inducible (Vgf) in the arcuate nucleus is involved in the seasonal regulation of energy balance. In situ hybridization, qPCR and RNA-sequencing studies have identified that Pomc expression across fish, avian and mammalian species, is a neuroendocrine marker that reflects seasonal energetic states. Here we highlight that long-term changes in arcuate Pomc and Vgf expression is conserved across species and may provide rheostatic regulation of seasonal energy balance. / Academy of Medical Sciences, Leverhulme Trust