The Nutritional Induction of COUP-TFII Gene Expression in Ventromedial Hypothalamic Neurons Is Mediated by the Melanocortin Pathway Lina Sabra-Makke1,2, Ce´ cile Tourrel-Cuzin3, Raphae¨ l G. P. Denis4, Marthe Moldes1,2, Jean-Paul Pe´ gorier1,2, Serge Luquet4, Mireille Vasseur-Cognet1,2*, Pascale Bossard1,2* 1 Institut Cochin, Universite´ Paris Descartes, Department of Endocrinology, Metabolism and Cancer, CNRS (UMR 8104), Paris, France, 2 INSERM, U1016, Paris, France, 3 Laboratoire de Biologie et Pathologie du Pancre´as Endocrine, Unite´ ‘‘Biologie Fonctionnelle & Adaptative’’, Universite´ Paris Diderot-Paris 7, CNRS EAC 4413, Paris, France, 4 Team HERGE, Unite´ ‘‘Functional and Adaptive Biology, Biologie Fonctionnelle & Adaptative’’, Universite´ Paris Diderot-Paris 7, CNRS EAC 4413, Paris, France Abstract Background: The nuclear receptor chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is an important coordinator of glucose homeostasis. We report, for the first time, a unique differential regulation of its expression by the nutritional status in the mouse hypothalamus compared to peripheral tissues. Methodology/Principal Findings: Using hyperinsulinemic-euglycemic clamps and insulinopenic mice, we show that insulin upregulates its expression in the hypothalamus. Immunofluorescence studies demonstrate that COUP-TFII gene expression is restricted to a subpopulation of ventromedial hypothalamic neurons expressing the melanocortin receptor. In GT1-7 hypothalamic cells, the MC4-R agonist MTII leads to a dose dependant increase of COUP-TFII gene expression secondarily to a local increase in cAMP concentrations. Transfection experiments, using a COUP-TFII promoter containing a functional cAMP responsive element, suggest a direct transcriptional activation by cAMP. Finally, we show that the fed state or intracerebroventricular injections of MTII in mice induce an increased hypothalamic COUP-TFII expression associated with a decreased hepatic and pancreatic COUP-TFII expression. Conclusions/Significance: These observations strongly suggest that hypothalamic COUP-TFII gene expression could be a central integrator of insulin and melanocortin signaling pathway within the ventromedial hypothalamus. COUP-TFII could play a crucial role in brain integration of circulating signal of hunger and satiety involved in energy balance regulation. Citation: Sabra-Makke L, Tourrel-Cuzin C, Denis RGP, Moldes M, Pe´gorier J-P, et al. (2010) The Nutritional Induction of COUP-TFII Gene Expression in Ventromedial Hypothalamic Neurons Is Mediated by the Melanocortin Pathway. PLoS ONE 5(10): e13464. doi:10.1371/journal.pone.0013464 Editor: Bernadette Breant, INSERM, France Received June 18, 2010; Accepted September 23, 2010; Published October 18, 2010 Copyright: 2010 Sabra-Makke et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by grants from ALFEDIAM-Antadir, France, 2008, and from the BQR (Bonus Qualite Recherche), Paris V University, France, 2007. L.S.M. is the recipient of a doctoral fellowship from the Conseil national de la recherche scientifique du Liban and the Fondation pour la Recherche Me´dicale. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: pascale.bossard@inserm.fr (PB); mireille.vasseur@inserm.fr (MV-C) Introduction The hypothalamus receives information from circulating nutri- ents and hormones and integration of these signals by appropriate neurocircuitry will in turn translate into adaptive metabolic and behavioral responses in order to sustain energy balance. Food intake modulation, insulin secretion and peripheral glucose metabolism together with endocrine regulation are part of this integrated response. Any disturbance of this mechanism leads to metabolic diseases such as obesity and diabetes [1,2]. Nuclear receptors are part of a family of ligand-activated transcription factor such as PPARs, LXRs, RXR, involved in the regulation of many key metabolic functions both at the central and peripheral level. Among these transcription factors, our team has focused its attention on the Chicken Ovalbumin Upstream Promoter-Transcription Factor II (COUP-TFII also named NR2F2). COUP-TFII is an orphan member of the steroid/ thyroid hormone receptor superfamily with no identified physio- logical ligand. It binds DNA by a zinc finger motif on a variety of hormone responsive elements that contain direct or inverted imperfect AGGTCA repeats with various spacings [3]. COUP- TFII is involved in the control of development, cellular differentiation, growth and metabolism [4,5,6]. In terms of metabolic control, we and others showed its functional role in respect to genetic programs associated with insulin biosynthesis and secretion in pancreatic beta cells [7,8], in the regulation of lipid utilization/cholesterol homeostasis in skeletal muscle cells [9] and in white adipose tissue development and energy metabolism [10]. The fact that heterozygous COUP-TFII gene inactivation mouse models, i.e conditional beta cells [8] or complete invalidation [10] (50% decrease in COUP-TFII expression) displayed a strong phenotype suggests that any small variations of COUP-TFII expression does lead to a disturbed physiology. Interestingly, its expression is modulated by the nutritional status in several tissues. We previously reported that COUP-TFII expression was reduced in the pancreas and liver of mice refed PLoS ONE | www.plosone.org 1 October 2010 | Volume 5 | Issue 10 | e13464