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Emma M Roberts
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Michael J F Newson
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George R Pope
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Rainer Landgraf Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, Max Planck Institute of Psychiatry, University of Bristol, Bristol BS1 3NY, UK

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Stephen J Lolait
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Anne-Marie O'Carroll
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2002 Apelin-immunoreactivity in the rat hypothalamus and pituitary . Neuroscience Letters 327 193 – 197 . Burbach JPH Luckman SM Murphy D Gainer H 2001 Gene-regulation in the magnocellular hypothalamo-neurohypophsial system

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M J F Newson Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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G R Pope Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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E M Roberts Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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S J Lolait Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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A-M O'Carroll Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology (LINE), School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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. Journal of Biological Chemistry 279 26274 – 26279 . ( doi:10.1074/jbc.M404149200 ) Jaszberenyi M Bujdoso E Telegdy G 2004 Behavioural, neuroendocrine and thermoregulatory actions of apelin-13 . Neuroscience 129 811 – 816 . ( doi:10

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Michael J F Newson Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Emma M Roberts Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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George R Pope Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Stephen J Lolait Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Anne-Marie O'Carroll Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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-immunoreactivity in the rat hypothalamus and pituitary . Neuroscience Letters 327 193 – 197 . Droste SK Chandramohan Y Hill LE Linthorst ACE Reul JMHM 2007 Voluntary exercise impacts on the rat hypothalamic–pituitary–adrenocortical axis mainly at

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L A Nolan Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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A Levy Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Neuroscience 2 894 – 897 . Candolfi M Zaldivar V Jaita G Seilicovich A 2006 Anterior pituitary cell renewal during the estrous cycle . Frontiers of Hormone Research 35 9 – 21 . Carbajo-Perez E Watanabe YG 1990 Cellular proliferation

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Yizhou Zhang Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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Meiqin Chen Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China

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Huan Chen Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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Shixiong Mi Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China

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Chang Wang Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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Hongchun Zuo Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China

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Leigang Song Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China

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Juan Du Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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Huixian Cui Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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Sha Li Department of Human Anatomy, Hebei Medical University, Shijiazhuang, Hebei, China
Neuroscience Research Center, Hebei Medical University, Shijiazhuang, Hebei, China
Hebei Key Laboratory of Neurodegenerative Disease Mechanism, Hebei Medical University, Shijiazhuang, Hebei, China

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feminization in the mouse . Nature 227 1217 – 1219 . ( https://doi.org/10.1038/2271217a0 ) Magee JC & Grienberger C 2020 Synaptic plasticity forms and functions . Annual Review of Neuroscience 43 95 – 117 . ( https://doi.org/10.1146/annurev

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Rui Gao Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, UK

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Samuel Acreman Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, UK
Department of Physiology, Institute of Neuroscience and Physiology, Metabolic Research Unit, University of Gothenburg, Göteborg, Sweden

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Jinfang Ma Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, UK

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Fernando Abdulkader Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, Brazil

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Anna Wendt Department of Clinical Sciences Malmö, Islet Cell Exocytosis, Lund University Diabetes Centre, Lund University, Malmö, Sweden

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Quan Zhang Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, UK
CNC - Center for Neuroscience and Cell Biology, CIBB - Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, Coimbra, Portugal

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Amperometric detection of single vesicle acetylcholine release events from an artificial cell . ACS Chemical Neuroscience 6 181 – 188 . ( https://doi.org/10.1021/cn5002667 ) Kimball CP & Murlin JR 1923 Aqueous extracts of pancreas: III . Journal of

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David M Golding School of Biosciences, Cardiff University, Cardiff, UK

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Daniel J Rees Institute of Life Sciences, College of Medicine, Swansea University, Swansea, UK

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Jennifer R Davies Behavioural Genetics Group, MRC Centre for Neuropsychiatric Genetics and Genomics, Neuroscience and Mental Health Research Institute, Schools of Medicine & Psychology, Cardiff University, Cardiff, UK

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Dinko Relkovic Behavioural Genetics Group, MRC Centre for Neuropsychiatric Genetics and Genomics, Neuroscience and Mental Health Research Institute, Schools of Medicine & Psychology, Cardiff University, Cardiff, UK

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Hannah V Furby Behavioural Genetics Group, MRC Centre for Neuropsychiatric Genetics and Genomics, Neuroscience and Mental Health Research Institute, Schools of Medicine & Psychology, Cardiff University, Cardiff, UK

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Irina A Guschina School of Biosciences, Cardiff University, Cardiff, UK

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Anna L Hopkins School of Biosciences, Cardiff University, Cardiff, UK

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Jeffrey S Davies Institute of Life Sciences, College of Medicine, Swansea University, Swansea, UK

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James L Resnick Center for Mammalian Genetics, University of Florida, College of Medicine, Gainesville, Florida, USA

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Anthony R Isles Behavioural Genetics Group, MRC Centre for Neuropsychiatric Genetics and Genomics, Neuroscience and Mental Health Research Institute, Schools of Medicine & Psychology, Cardiff University, Cardiff, UK

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Timothy Wells School of Biosciences, Cardiff University, Cardiff, UK

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Neuroscience 42 2105 – 2113 . ( doi:10.1111/ejn.12972 ) Davies JS Kotokorpi P Eccles SR Barnes SK Tokarczuk PF Allen SK Whitworth HS Guschina IA Evans BA Mode A 2009 Ghrelin induces abdominal obesity via GHS

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K L Davies Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK

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E J Camm Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK
The Ritchie Centre, Hudson Institute of Medical Research, Clayton, Australia

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D J Smith Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK

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O R Vaughan Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK
Institute for Women’s Health, University College London, London, UK

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A J Forhead Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK
Department of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK

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A J Murray Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK

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A L Fowden Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK

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authors thank the staff of the University Biomedical Services for their care of the animals and the technical staff of the Department of Physiology, Development and Neuroscience who assisted with this study. References Arvier M Langoutte L

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K L Davies Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK

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J Miles Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK

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E J Camm Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK
The Ritchie Centre, Hudson Institute of Medical Research, Clayton, Australia

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D J Smith Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK

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P Barker MRC Metabolic Diseases Unit, Mouse Biochemistry Laboratory, Cambridge Biomedical Campus, Cambridge, UK

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K Taylor Endocrine Laboratory, Blood Sciences, Cambridge University Hospitals NHS Foundation Trust, Hills Road, Cambridge, UK

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A J Forhead Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK
Department of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK

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A L Fowden Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge, UK

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hypothalamic-pituitary-adrenal axis: development, programming actions of hormones and maternal-fetal interactions . Frontiers in Behavioral Neuroscience 14 601939 . ( https://doi.org/10.3389/fnbeh.2020.601939 ) Sloboda DM Moss TJM Gurrin LC Challis

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Ann R Finch Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Kathleen R Sedgley Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Christopher J Caunt Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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Craig A McArdle Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK

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In heterologous expression systems, human GnRH receptors (hGnRHRs) are poorly expressed at the cell surface and this may reflect inefficient exit from the endoplasmic reticulum. Here, we have defined the proportion of GnRHRs at the cell surface using a novel assay based on adenoviral transduction with epitope-tagged GnRHRs followed by staining and semi-automated imaging. We find that in MCF7 (breast cancer) cells, the proportional cell surface expression (PCSE) of hGnRHRs is remarkably low (<1%), when compared with Xenopus laevis (X) GnRHRs (∼40%). This distinction is retained at comparable whole cell expression levels, and the hGnRHR PCSE is increased by addition of the XGnRHR C-tail (h.XGnRHR) or by a membrane-permeant pharmacological chaperone (IN3). The IN3 effect is concentration- and time-dependent and IN3 also enhances the hGnRHR-mediated (but not h.XGnRHR- or mouse GnRHR-mediated) stimulation of [3H]inositol phosphate accumulation and the hGnRHR-mediated reduction in cell number. We also find that the PCSE for hGnRHRs and h.XGnRHRs is low and is greatly increased by IN3 in two hormone-dependent cancer lines, but is higher and less sensitive to IN3 in a gonadotrope line. Finally, we show that the effect of IN3 on hGnRHR PCSE is not mimicked or blocked by two peptide antagonists although they do increase the PCSE for h.XGnRHRs, revealing that an antagonist-occupied cell surface GnRHR conformation can differ from that of the unoccupied receptor. The low PCSE of hGnRHRs and this novel peptide antagonist effect may be important for understanding GnRHR function in extrapituitary sites.

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