The Journal of Clinical Endocrinology & Metabolism, 2021, Vol. 106, No. 2, e534–e550 doi:10.1210/clinem/dgaa782 Clinical Research Article e534 https://academic.oup.com/jcem ISSN Print 0021-972X ISSN Online 1945-7197 Printed in USA © The Author(s) 2020. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com Clinical Research Article A Novel Mutation in the FSH Receptor (I423T) Affecting Receptor Activation and Leading to Primary Ovarian Failure Teresa Zariñán, 1 Julio Mayorga, 2 Eduardo Jardón-Valadez, 3 Rubén Gutiérrez-Sagal, 1 José Luis Maravillas-Montero, 1 Nancy R. Mejía-Domínguez, 1 Iván Martínez-Luis, 1 Omar G. Yacini-Torres 1 , Ma.-del-Carmen Cravioto, 2 Eric Reiter, 4 and Alfredo Ulloa-Aguirre 1 1 Red de Apoyo a la Investigación, National University of Mexico-Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico; 2 Department of Reproductive Biology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico; 3 Departamento de Recursos de la Tierra, Universidad Autónoma Metropolitana, Unidad Lerma, Lerma, Edo. de Mexico, Mexico; and 4 Physiologie de la Reproduction et des Comportements (PRC), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre National de la Recherche Scientifique (CNRS), Université de Tours, Tours, France ORCiD numbers: 0000-0003-3701-429X (E. Jardón-Valadez); 0000-0003-3755-662X (J. L. Maravillas-Montero); 0000-0002-9532- 3886 (A. Ulloa-Aguirre). Abbreviations: AUC, areas under the curve; BSA, bovine serum albumin; cAMP, cyclic adenosine monophosphate; DNA, deoxyribonucleic acid; ECD, extracellular domain; FSH, follicle-stimulating hormone; FSHR, follicle-stimulating hormone receptor; HBSD, hormone binding specific domain; IBMX, 3-isobutyl-methyl-xanthine; Ig, immunoglobulin; LH, luteinizing hormone; LHCGR, luteinizing hormone choriogonadotropin receptor; MSF, mean square fluctuation; PBS, phosphate- buffered saline; PC, principal components; PCA, principal components analysis; PCR, polymerase chain reaction; PKA, protein kinase A; PM, plasma membrane; POF, primary ovarian failure; RIA, radioimmunoassay; RMSF, root mean square fluctuation; TMD, transmembrane helical domain; WT, wild type. Received: 9 September 2020; Editorial Decision: 19 October 2020; First Published Online: 29 October 2020; Corrected and Typeset: 3 December 2020. Abstract Context: Follicle-stimulating hormone (FSH) plays an essential role in gonadal function. Loss-of-function mutations in the follicle-stimulating hormone receptor (FSHR) are an infrequent cause of primary ovarian failure. Objective: To analyze the molecular physiopathogenesis of a novel mutation in the FSHR identifed in a woman with primary ovarian failure, employing in vitro and in silico approaches, and to compare the features of this dysfunctional receptor with those shown by the traffcking-defective D408Y FSHR mutant. Methods: Sanger sequencing of the FSHR cDNA was applied to identify the novel mutation. FSH-stimulated cyclic adenosine monophosphate (cAMP) production, ERK1/2 phosphorylation, and desensitization were tested in HEK293 cells. Receptor expression was analyzed by immunoblotting, receptor-binding assays, and fow cytometry. Molecular Downloaded from https://academic.oup.com/jcem/article/106/2/e534/5942952 by guest on 17 June 2022