Y1 and Y5 Receptors Are Both Required for the Regulation of Food Intake and Energy Homeostasis in Mice

Neuropeptide Y (NPY) acting in the hypothalamus is one of the most powerful orexigenic agents known. Of the five known Y receptors, hypothalamic Y1 and Y5 have been most strongly implicated in mediating hyperphagic effects. However, knockout of individual Y1 or Y5 receptors induces late-onset obesity – and Y5 receptor knockout also induces hyperphagia, possibly due to redundancy in functions of these genes. Here we show that food intake in mice requires the combined actions of both Y1 and Y5 receptors. Germline Y1Y5 ablation in Y1Y5−/− mice results in hypophagia, an effect that is at least partially mediated by the hypothalamus, since mice with adult-onset Y1Y5 receptor dual ablation targeted to the paraventricular nucleus (PVN) of the hypothalamus (Y1Y5Hyp/Hyp) also exhibit reduced spontaneous or fasting-induced food intake when fed a high fat diet. Interestingly, despite hypophagia, mice with germline or hypothalamus-specific Y1Y5 deficiency exhibited increased body weight and/or increased adiposity, possibly due to compensatory responses to gene deletion, such as the decreased energy expenditure observed in male Y1Y5−/− animals relative to wildtype values. While Y1 and Y5 receptors expressed in other hypothalamic areas besides the PVN – such as the dorsomedial nucleus and the ventromedial hypothalamus – cannot be excluded from having a role in the regulation of food intake, these studies demonstrate the pivotal, combined role of both Y1 and Y5 receptors in the mediation of food intake

NPY Neuron-Specific Y2 Receptors Regulate Adipose Tissue and Trabecular Bone but Not Cortical Bone Homeostasis in Mice

BACKGROUND: Y2 receptor signalling is known to be important in neuropeptide Y (NPY)-mediated effects on energy homeostasis and bone physiology. Y2 receptors are located post-synaptically as well as acting as auto receptors on NPY-expressing neurons, and the different roles of these two populations of Y2 receptors in the regulation of energy homeostasis and body composition are unclear. METHODOLOGY/PRINCIPAL FINDINGS: We thus generated two conditional knockout mouse models, Y2(lox/lox) and NPYCre/+;Y2(lox/lox), in which Y2 receptors can be selectively ablated either in the hypothalamus or specifically in hypothalamic NPY-producing neurons of adult mice. Specific deletion of hypothalamic Y2 receptors increases food intake and body weight compared to controls. Importantly, specific ablation of hypothalamic Y2 receptors on NPY-containing neurons results in a significantly greater adiposity in female but not male mice, accompanied by increased hepatic triglyceride levels, decreased expression of liver carnitine palmitoyltransferase (CPT1) and increased expression of muscle phosphorylated acetyl-CoA carboxylase (ACC). While food intake, body weight, femur length, bone mineral content, density and cortical bone volume and thickness are not significantly altered, trabecular bone volume and number were significantly increased by hypothalamic Y2 deletion on NPY-expressing neurons. Interestingly, in situ hybridisation reveals increased NPY and decreased proopiomelanocortin (POMC) mRNA expression in the arcuate nucleus of mice with hypothalamus-specific deletion of Y2 receptors in NPY neurons, consistent with a negative feedback mechanism between NPY expression and Y2 receptors on NPY-ergic neurons. CONCLUSIONS/SIGNIFICANCE: Taken together these data demonstrate the anti-obesogenic role of Y2 receptors in the brain, notably on NPY-ergic neurons, possibly via inhibition of NPY neurons and concomitant stimulation of POMC-expressing neurons in the arcuate nucleus of the hypothalamus, reducing lipogenic pathways in liver and/or skeletal muscle in females. These data also reveal as an anti-osteogenic effect of Y2 receptors on hypothalamic NPY-expressing neurons on trabecular but not on cortical bone

Greater bone formation of Y2 knockout mice is associated with increased osteoprogenitor numbers and altered Y1 receptor expression

Germ line or hypothalamus-specific deletion of Y2 receptors in mice results in a doubling of trabecular bone volume. However, the specific mechanism by which deletion of Y2 receptors increases bone mass has not yet been identified. Here we show that cultured adherent bone marrow stromal cells from Y2(-/-) mice also demonstrate increased mineralization in vitro. Isolation of two populations of progenitor cell types, an immature mesenchymal stem cell population and a more highly differentiated population of progenitor cells, revealed a greater number of the progenitor cells within the bone of Y2(-/-) mice. Analysis of Y receptor transcripts in cultured stromal cells from wild-type mice revealed high levels of Y1 but not Y2, Y4, Y5, or y6 receptor mRNA. Interestingly, germ line Y2 receptor deletion causes Y1 receptor down-regulation in stromal cells and bone tissue possibly due to the lack of feedback inhibition of NPY release and subsequent overstimulation of Y1 receptors. Furthermore, deletion of Y1 receptors resulted in increased bone mineral density in mice. Together, these findings indicate that the greater number of mesenchymal progenitors and the altered Y1 receptor expression within bone cells in the absence of Y2 receptors are a likely mechanism for the greater bone mineralization in vivo and in vitro, opening up potential new treatment avenues for osteoporosis

Novel role of Y1 receptors in coordinated regulation of bone and energy homeostasis

The importance of neuropeptide Y (NPY) and Y2 receptors in the regulation of bone and energy homeostasis has recently been demonstrated. However, the contributions of the other Y receptors are less clear. Here we show that Y1 receptors are expressed on osteoblastic cells. Moreover, bone and adipose tissue mass are elevated in Y1-/- mice with a generalized increase in bone formation on cortical and cancellous surfaces. Importantly, the inhibitory effects of NPY on bone marrow stromal cells in vitro are absent in cells derived from Y1-/- mice, indicating a direct action of NPY on bone cells via this Y receptor. Interestingly, in contrast to Y2 receptor or germ line Y1 receptor deletion, conditional deletion of hypothalamic Y1 receptors in adult mice did not alter bone homeostasis, food intake, or adiposity. Furthermore, deletion of both Y1 and Y2 receptors did not produce additive effects in bone or adiposity. Thus Y1 receptor pathways act powerfully to inhibit bone production and adiposity by nonhypothalamic pathways, with potentially direct effects on bone tissue through a single pathway with Y2 receptors

Altered function of arcuate leptin receptor expressing neuropeptide Y neurons depending on energy balance

Objective: One of leptin's main targets in the hypothalamus are neuropeptide Y (NPY) neurons, with selective deletion of leptin receptors (Lepr) specifically in Npy neurons resulting in major alterations of energy partitioning between fat and bone mass. However, the specific action of these Npy+/Lepr+ neurons compared to Npy-negative Lepr (Npy−/Lepr+) neurons in regard to energy homeostasis regulation is unknown. Methods: Specific AAV viral vectors were generated using DREADD and INTRSECT technology and used in male LeprCre/+ and LeprCre/+;NpyFlp/+ mice to assess the effect of activating either all Lepr neurons or specifically Npy+/Lepr+ or Npy−/Lepr+ neurons only on feeding, energy homeostasis control, and body composition. Results: Selective stimulation of Npy+/Lepr+ neurons led to an immediate decrease in respiratory quotient followed by a delayed increase in food intake in standard chow fed, but interestingly not in high fat diet (HFD) fed mice. In addition, stimulation of Npy+/Lepr+ neurons led to a robust increase in brown adipose tissue thermogenesis and improved glucose tolerance. These effects were not observed in standard chow fed mice when Npy−/Lepr+ expressing neurons were specifically activated, suggesting the effects of leptin on these parameters are driven by NPY. However, under HFD condition when leptin levels are elevated, the stimulation of the Npy−/Lepr+ neurons increased food intake, physical activity and energy expenditure. Interestingly, chronic stimulation of Npy-positive Lepr neurons was able to increase bone mass independently of bodyweight, whilst chronic stimulation of the Npy−/Lepr+ neurons resulted in increased bodyweight and fat mass with proportionate increases in bone mass. Conclusions: Together, these data indicate that leptin signalling through Npy-positive Lepr-expressing neurons controls energy partitioning via stimulation of thermogenesis, energy expenditure, and the use of fat as a fuel source. However, under prolonged HFD, leptin resistance may occur and actions of leptin signalling through Npy-negative Lepr hypothalamic neurons may exacerbate excess food intake

Synergistic Effects of Y2 and Y4 Receptors on Adiposity and Bone Mass Revealed in Double Knockout Mice

Neuropeptide Y regulates numerous physiological processes via at least five different Y receptors, but the specific roles of each receptor are still unclear. We previously demonstrated that Y2 receptor knockout results in a lean phenotype, increased cancellous bone volume, and an increase in plasma pancreatic polypeptide (PP), a ligand for Y4 receptors. PP-overexpressing mice are also known to have a lean phenotype. Deletion of the Y4 receptor also produced a lean phenotype and increased plasma PP levels. We therefore hypothesized that part of the Y2 phenotype results from increased PP action on Y4 receptors and tested this in PP transgenic Y4(−/−) and Y2(−/−) Y4(−/−) double knockout mice. Bone mass was not altered in Y4 knockout mice. Surprisingly, despite significant hyperphagia, Y2(−/−) Y4(−/−) mice retained a markedly lean phenotype, with reduced body weight, white adipose tissue mass, leptinemia, and insulinemia. Furthermore, bone volume was also increased threefold in Y2(−/−) Y4(−/−) mice, and this was associated with enhanced osteoblastic activity. These changes were more pronounced than those observed in Y2(−/−) mice, suggesting synergy between Y2 and Y4 receptor pathways. The lack of bone changes in PP transgenic mice suggests that PP alone is not responsible for the bone mass increases but might play a major role in the lean phenotype. However, a synergistic interaction between Y2 and Y4 pathways seems to regulate bone volume and adiposity and could have important implications for possible interventions in obesity and for anabolic treatment of osteoporotic bone loss

Critical interplay between neuropeptide Y and sex steroid pathways in bone and adipose tissue homeostasis

Synchronous languages are used to program critical reactive systems. Today, systems require to find a way to execute them safely and in parallel. Parallelism has always been part of synchronous langages, but for modeling purpose. Their formal semantics allow to distribute them, but preserving the semantics may be ressource costly and prevent good parallel execution.The Kahn networks model is of great interest. It models distributed computers, communicating through unbounded FIFOs, ensuring that the computed values are deterministic, without any need of added synchronization.We develop the langage Heptagon, a first order functional synchronous son of Lustre.The compiler is an academic prototype of the industrial tool Scade. Thanks to its Kahn semantics, it can be distributed. In order to be efficient, one need to maximize the decoupling of computations and maximize the computation granularity. However, synchronous langages allow for very tight computation coupling and usually require thin computation granularity to ensure reactivity of the system.We opt for two research directions. The first one is to give the control of the execution parallelism to the programer. To this mean, we add futures to the source langage Heptagon. They provide control over starting and end of parallel computations, while preserving the functional semantics. Moreover, we provide a compilation for embedded systems, using statically allocated memory. The second one is to study Kahn synchronous semantics to understand data dependencies and maximize granularity of the computations. This touches deeply to the synchronous languages, mixing the usually separated questions of causality and clock calculus. We define the class of reactive ordered Kahn networks. They are the one which may be modularly compiled and whose behavior may be expressed with a clock signature. Moreover, we show that their is a normal form for this signature, maximizing the granularity of the network. To express it, we extend clocks to integer clocks. Then we come back to the synchronous languages we know to understand how to use it. The result is fully used and explained on Lucy-n, the synchronous language closest to Kahn networks.Les langages synchrones ont été fondés pour modéliser et implémenter les systèmes réactifs temps-réels critiques. Avec la complexité toujours croissante des systèmes contrôlés, la vitesse d'exécution devient un critère important. Nous sommes donc à la recherche d'une exécution parallèle, combinant efficacité et sûreté.Les langages synchrones ont toujours intégré la notion de parallélisme, mais ce, pour l'expressivité de la modélisation. Leurs compilations visent principalement les circuits ou la génération de code séquentiel. Tous ont une sémantique formelle, qui rend possible la distribution correcte du code. Mais la préservation de cette sémantique peut être un obstacle à l'efficacité du code généré, particulièrement s'il est nécessaire de préserver une notion d'instant global au système.Le modèle sémantique qui nous intéresse est celui des réseaux de Kahn. Ces réseaux modélisent des calculateurs distribués, communiquant au travers de files de taille non bornée. Dans ce cadre, la distribution ne demande aucune communication ni synchronisation supplémentaire. En considérant l'histoire des files de communication, la sémantique de Kahn permet de s'abstraire de l'exécution effective, tout en garantissant le déterminisme du calcul. Pour cela, chaque nœud du réseau doit avoir une sémantique fonctionnelle continue.Le langage que nous développons est Heptagon, un langage synchrone fonctionnel du premier ordre, déscendant de Lustre. Son compilateur est un prototype universitaire, apparenté à l'outil industriel Scade. Grâce à sa sémantique de Kahn, la distribution d'un programme Heptagon ne pose pas de question, son efficacité beaucoup plus.L'efficacité requiert de minimiser les synchronisations. Cela revêt deux aspects non indépendants. Avoir un découplage suffisant des calculs : il y a des délais dans les dépendances entre calculs. Avoir une granularité importante des calculs : un fort ratio temps de calcul sur fréquence de communication. Or la sémantique synchrone et les horloges d'un programme Heptagon reflètent exactement l'inverse. Elles permettent au programmeur de se contenter d'un découplage d'un instant et à chaque instant, au maximum une valeur est calculée. De plus, les instants sont typiquement courts, pour assurer que le système réagit rapidement.Des précédents travaux sur le sujet, nous tirons deux constats.Le premier est que nous souhaitons le contrôle du parallélisme par le programmeur, directement dans le code source. Il doit pouvoir maîtriser à quels instants il y a communication ou synchronisation. La solution que nous proposons dans ce manuscrit est l'utilisation des futures dans Heptagon. Ils fournissent ce pouvoir au programmeur, tout en restant des annotations qui peuvent être supprimées sans changer la sémantique dénotationnelle du programme.Le deuxième constat est que la question de la granularité des calculs est une question profonde, touchant en particulier aux questions de dépendance de données, de choix des horloges et de compilation modulaire. Heptagon, comme ses parents, restreint les réseaux de Kahn qui peuvent être écrits, de telle sorte que ces trois questions se traitent séparément. Pour mieux comprendre le lien entre ces éléments, nous revenons aux réseaux de Kahn. Notre principal résultat est la définition de la sous-classe des réseaux ordonnés réactifs. Ceux-ci sont les seuls pour lesquels nous pouvons décrire modulairement le comportement avec des horloges, sans restreindre les contextes d'appels. Ces réseaux ont une signature d'horloge en forme normale, qui maximise la granularité. Pour l'exprimer, nous introduisons les horloges entières, décrivant la communication de plusieurs valeurs en un seul instant. Nous appliquons ensuite nos résultats pour voir sous un nouveau jour Heptagon, Signal, les politiques des objets de Lucid Synchrone, mais aussi proposer une analyse pleinement modulaire de Lucy-n langage synchrone le plus fidèle aux réseaux de Kahn