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Fabio V Comim Imperial College London, Institute of Biodiversity, Institute of Reproductive and Developmental Biology, London W12 0NN, UK

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Kate Hardy Imperial College London, Institute of Biodiversity, Institute of Reproductive and Developmental Biology, London W12 0NN, UK

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Jane Robinson Imperial College London, Institute of Biodiversity, Institute of Reproductive and Developmental Biology, London W12 0NN, UK

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Stephen Franks Imperial College London, Institute of Biodiversity, Institute of Reproductive and Developmental Biology, London W12 0NN, UK

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polycystic ovaries ( Webber et al . 2003 , 2007 , Stubbs et al . 2007 ), these findings support the concept that aberrant follicle development in PCOS is programmed by exposure of the ovary to excess androgen during foetal life. Declaration of interest

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S E Chadio Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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B Kotsampasi Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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G Papadomichelakis Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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S Deligeorgis Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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D Kalogiannis Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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I Menegatos Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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G Zervas Laboratory of Anatomy and Physiology of Domestic Animals,
Laboratory of Animal Nutrition,
Laboratory of Animal Breeding, Department of Animal Science, Agricultural University of Athens, 75, Iera odos, 11855 Athens, Greece

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Introduction A series of experimental and epidemiological studies had led to the fetal programming hypothesis, which implies that adverse environmental factors, acting in utero program the development of fetal tissues, producing

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Hao Wu Department of Nephrology, The Second Hospital of Jilin University, Changchun, Jilin, People’s Republic of China
The ‘973’ National Basic Research Program of China, Changchun University of Chinese Medicine, Changchun, Jilin, People’s Republic of China

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Junduo Wu Department of Cardiology, The Second Hospital of Jilin University, Changchun, Jilin, People’s Republic of China

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Shengzhu Zhou Department of Anesthesiology, The Second Hospital of Jilin University, Changchun, Jilin, People’s Republic of China

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Wenlin Huang School of Science and Technology, Georgia Gwinnett College, Lawrenceville, Georgia, USA

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Ying Li Department of Dermatology, Affiliated Hospital of Beihua University, Jilin, Jilin, People’s Republic of China

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Huan Zhang Operating Theatre, China-Japan Union Hospital of Jilin University, Changchun, Jilin, People’s Republic of China

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Junnan Wang Department of Cardiology, The Second Hospital of Jilin University, Changchun, Jilin, People’s Republic of China

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Ye Jia Department of Nephrology, The First Hospital of Jilin University, Changchun, Jilin, People’s Republic of China

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interest that could be perceived as prejudicing the impartiality of the research reported. Funding This work was supported in part by the National Natural Science Foundation of China (81600573) and Norman Bethune Program of Jilin University (2015438

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L Johnsen Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Groennegaardsvej 7, DK-1870 Frederiksberg, Denmark

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A H Kongsted Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Groennegaardsvej 7, DK-1870 Frederiksberg, Denmark

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M O Nielsen Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Groennegaardsvej 7, DK-1870 Frederiksberg, Denmark

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exposures during fetal life with altered body functions and health outcomes later in life is often referred to as fetal metabolic programing (FMP). Experience from developing countries undergoing rapid economic transition indicates that individuals who have

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Kazunari Nohara Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Suhuan Liu Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Matthew S Meyers Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Aurélie Waget Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Mathieu Ferron Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Gérard Karsenty Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Rémy Burcelin Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Franck Mauvais-Jarvis Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA
Division of Endocrinology, Division of Endocrinology, Department of Genetics and Development, INSERM U1048, Metabolism and Molecular Medicine and Comprehensive Center on Obesity, Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA

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Introduction Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in females of reproductive age and is believed to have a developmental origin in which maternal and fetal androgen excess during pregnancy programs both the

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Anne-Lise Lecoq Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France

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Philippe Zizzari Inserm U894, Centre de Psychiatrie et Neurosciences, Université Paris Descartes, Sorbonne Paris Cité, Paris, France

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Mirella Hage Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France

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Lyvianne Decourtye Sorbonne Universités, Univ Paris 06 UMRS 938, Inserm U938, CDR Saint-Antoine, Paris, France

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Clovis Adam Assistance Publique-Hôpitaux de Paris, Service d’Anatomie et Cytologie Pathologiques, Hôpital Bicêtre, Le Kremlin Bicêtre, France

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Say Viengchareun Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France

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Johannes D Veldhuis Department of Medicine, Endocrine Research Unit, Mayo School of Graduate Medical Education, Clinical Translational Science Center, Mayo Clinic, Rochester, Minnesota, USA

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Valérie Geoffroy Inserm U1132, Hôpital Lariboisière, Université Paris Diderot, Sorbonne Paris Cité, Paris, France

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Marc Lombès Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France

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Virginie Tolle Inserm U894, Centre de Psychiatrie et Neurosciences, Université Paris Descartes, Sorbonne Paris Cité, Paris, France

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Anne Guillou Unité Mixte de Recherche-5203, Centre National de la Recherche Scientifique, Institut de Génomique Fonctionnelle, Montpellier, France

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Auli Karhu Department of Medical Genetics, Genome-Scale Biology Research Program Biomedicum, University of Helsinki, Helsinki, Finland

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Laurent Kappeler Sorbonne Universités, Univ Paris 06 UMRS 938, Inserm U938, CDR Saint-Antoine, Paris, France

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Philippe Chanson Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France
Assistance Publique-Hôpitaux de Paris, Service d’Endocrinologie et des Maladies de la Reproduction, Hôpital Bicêtre, Le Kremlin Bicêtre, France

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Peter Kamenický Institut National de la Santé et de la Recherche Médicale (Inserm) U1185, Le Kremlin-Bicêtre, France
Université Paris-Sud, Faculté de Médecine Paris-Sud, Le Kremlin-Bicêtre, France
Assistance Publique-Hôpitaux de Paris, Service d’Endocrinologie et des Maladies de la Reproduction, Hôpital Bicêtre, Le Kremlin Bicêtre, France

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VECTOR M.O.M. Immunodetection Kit FMK-2201 (Vector Laboratories, Eurobio) and the Dako Biotin Blocking System (X0590, Dako). Slides were incubated with a rabbit anti-GH antibody 1:20,000 (AFP5641801, NIDDK, National Hormone and Peptide Program

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Bo He Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, China
School of Pharmaceutical Sciences and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming, China

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Yinxian Wen Department of Orthopedic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China
Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, China

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Shuwei Hu Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, China

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Guihua Wang Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, China

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Wen Hu Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, China

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Jacques Magdalou UMR 7561 CNRS-Université de Lorraine, Faculté de Médicine, Vandoeuvre-lès-Nancy, Nancy, France

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Liaobin Chen Department of Orthopedic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China
Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, China

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Hui Wang Department of Pharmacology, Wuhan University School of Basic Medical Sciences, Wuhan, China
Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan, China

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have demonstrated that PCE can lead to fetal overexposure to maternal glucocorticoids and hypothalamic-pituitary-adrenal (HPA) axis-associated neuroendocrine metabolic programming alterations, resulting in the increased susceptibility of adult offspring

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Rob H P Hilgers
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Suzanne Oparil
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Wout Wouters Division of Cardiovascular Disease, Pantarhei Bioscience, Department of Medicine, Vascular Biology and Hypertension Program, University of Alabama at Birmingham, 1034 Zeigler Research Building, 703 19th Street South, Birmingham, Alabama 35294-0007, USA

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Herjan J T Coelingh Bennink Division of Cardiovascular Disease, Pantarhei Bioscience, Department of Medicine, Vascular Biology and Hypertension Program, University of Alabama at Birmingham, 1034 Zeigler Research Building, 703 19th Street South, Birmingham, Alabama 35294-0007, USA

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This study compared ex vivo relaxing responses to the naturally occurring human hormone estetrol (E4) vs 17β-estradiol (E2) in eight different vascular beds. Arteries were mounted in a myograph, contracted with either phenylephrine or serotonin, and cumulative concentration-response curves (CRCs) to E4 and E2 (0.1–100 μmol/l) were constructed. In all arteries tested, E4 had lower potency than E2, although the differential effect was less in larger than smaller arteries. In uterine arteries, the nonselective estrogen receptor (ER) blocker ICI 182 780 (1 μmol/l) caused a significant rightward shift in the CRC to both E4 and E2, indicating that the relaxation responses were ER dependent. Pharmacological blockade of nitric oxide (NO) synthases by N ω-nitro-l-arginine methyl ester (l-NAME) blunted E2-mediated but not E4-mediated relaxing responses, while inhibition of prostaglandins and endothelium-dependent hyperpolarization did not alter relaxation to either E4 or E2 in uterine arteries. Combined blockade of NO release and action with l-NAME and the soluble guanylate cyclase (sGC) inhibitor ODQ resulted in greater inhibition of the relaxation response to E4 compared with E2 in uterine arteries. Endothelium denudation inhibited responses to both E4 and E2, while E4 and E2 concentration-dependently blocked smooth muscle cell Ca2 + entry in K+-depolarized and Ca2 +-depleted uterine arteries. In conclusion, E4 relaxes precontracted rat arteries in an artery-specific fashion. In uterine arteries, E4-induced relaxations are partially mediated via an endothelium-dependent mechanism involving ERs, sGC, and inhibition of smooth muscle cell Ca2 + entry, but not NO synthases or endothelium-dependent hyperpolarization.

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Chalida Nakalekha Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Chieko Yokoyama Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Hiroyuki Miura Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Neil Alles Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Kazuhiro Aoki Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Keiichi Ohya Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Ikuo Morita Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School
Departments of, Cellular Physiological Chemistry, Fixed Prosthodontics, International Research Center for Molecular Science in Tooth and Bone Diseases (Global COE program), Institute of Cellular and System Medicine, Department of Hard Tissue Engineering (Pharmacology), Graduate School

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Prostaglandins (PGs) are key regulatory factors that affect bone metabolism. Prostaglandin E2 (PGE2) regulates bone resorption and bone formation. Prostacyclin (PGI2) is one of the major products derived from arachidonic acid by the action of cyclooxygenase and PGI2 synthase (PGIS). Unlike PGE2, there are few reports about the role of PGI2 in bone regulation. Therefore, we investigated the potential effect of PGI2 on bone metabolism. We used PGIS knockout (PGIS−/−), PGIS heterozygous (PGIS+ /−), and wild-type mice to investigate the role of PGI2. Notably, PGIS−/− mice gradually displayed an increase in trabecular bone mass in adolescence. Adult PGIS−/− mice showed an increase in trabecular bone volume/tissue volume. Histomorphometric analysis showed that PGIS−/− mice displayed increases in both bone formation and bone resorption parameters. Levels of serum osteocalcin and C-telopeptides were increased in adult PGIS−/− mice. Furthermore, the increased bone mass patterns were rescued in PGIS/tg mice. In conclusion, adult PGIS−/− mice displayed an overall increase in the levels of both bone formation and bone resorption parameters, which suggests that PGI2 deficiency accelerates high bone turnover activity with a greater increase in bone mass in aging. These results indicated that PGI2 may contribute to the maintenance of normal bone mass and micro-architecture in mice in age-dependent manner. Our findings demonstrate for the first time that PGI2 is involved in bone metabolism in vivo.

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Virginia L Pszczolkowski Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Endocrinologh and Reproductive Physiology Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

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Meghan K Connelly Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Vita Plus Corporation, Madison, Wisconsin, USA

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Adam D Beard Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Endocrinologh and Reproductive Physiology Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

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Amara D Benn Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA

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Jimena Laporta Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Endocrinologh and Reproductive Physiology Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

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Laura L Hernandez Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Endocrinologh and Reproductive Physiology Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

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Sebastian I Arriola Apelo Department of Animal and Dairy Sciences, University of Wisconsin, Madison, Wisconsin, USA
Endocrinologh and Reproductive Physiology Graduate Training Program, University of Wisconsin, Madison, Wisconsin, USA

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Energy partitioning in lactating cows affects milk production, feed efficiency, and body reserves, with the latter having health implications for the transition into the following lactation. One molecule likely involved in the regulation of energy partitioning is serotonin. The objective of this experiment was to explore how increasing circulating serotonin, by intravenous infusion of the serotonin precursor 5-hydroxytryptophan (5-HTP), affects metabolic responses to a glucose challenge in midlactation cows as a means to manipulate energy partitioning. We intravenously infused Holstein cows with 5-HTP (1 mg/kg bodyweight dissolved in saline, n = 11) or saline alone as control (n = 9) over 1 h/day for 3 days. Cows were fasted overnight on day 2. On day 3, fasted cows were given an intravenous bolus of glucose (0.092 g/kg bodyweight). Blood samples were collected for the following 120 min for metabolic and hormonal analysis. Infusion of 5-HTP elevated circulating concentrations of serotonin and free fatty acids, reduced the concentration of insulin and amino acids, and did not affect the concentration of glucose and glucagon before the glucose challenge. Surrogate insulin sensitivity indices indicated improved insulin sensitivity in 5-HTP cows, but due to the unique metabolism of lactating ruminants, these index changes may instead reflect effects in insulin-independent glucose disposal, like milk synthesis. Challenging 5-HTP-treated cows with a glucose bolus reduced the insulin spike and blunted the decrease in free fatty acids, compared to saline cows, without changing glucose dynamics. Overall, these results suggest that serotonin stimulates insulin-independent glucose disposal, requiring less insulin to maintain normoglycemia.

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