title: MicroRNA miR-513a-3p acts as a co-regulator of luteinizing hormone/chorionic gonadotropin receptor gene expression in human granulosa cells.
abstract: The luteinizing hormone/chorionic gonadotropin receptor (LHCGR) is essential for normal male and female reproductive processes. The spatial and temporal LHCGR gene expression is controlled by a complex system of regulatory mechanisms which are crucial for normal physiological function, especially during the female cycle. In this study, we aimed to elucidate whether microRNAs are involved in this network and play a role in regulating LHCGR expression. Computational analysis predicted that miR-513a-3p interacts with the LHCGR mRNA via three binding sites located in the 3'UTR region, enabling a synergistic action. Moreover, using a luciferase-based reporter assay we found that miR-513a-3p targets the LHCGR, resulting in a significant down-regulation of its expression. In human primary granulosa cell cultures we detected a dynamic, inversely associated expression pattern of miR-513a-3p and the LHCGR. In addition, transfection with miR-513a-3p or its specific inhibitor led to a down- or up-regulation at the LHCGR mRNA level, respectively. An increased amount of miR-513a-3p resulted in the down-regulation of the LHCGR mRNA, reflected by the attenuation of cAMP synthesis after hormonal stimulation. In conclusion, these data demonstrate that miR-513a-3p is involved in the control of the LHCGR expression by an inversely regulated mechanism at the post-transcriptional level and show for the first time that this kind of post-transcriptional process contributes to the multifaceted system of the human LHCGR regulation.
authors.affiliation: Centre of Reproductive Medicine and Andrology, University Hospital Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Troppmann
authors.affiliation: Centre of Reproductive Medicine and Andrology, University Hospital Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Kossack
authors.affiliation: Centre of Reproductive Medicine and Andrology, University Hospital Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Nordhoff
authors.affiliation: Department of Gynaecology and Obstetrics, University Hospital Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany.
authors.forename: A N
authors.initials: AN
authors.lastname: Schüring
authors.affiliation: Centre of Reproductive Medicine and Andrology, University Hospital Muenster, Albert-Schweitzer-Campus 1, 48149 Muenster, Germany. Electronic address: joerg.gromoll@ukmuenster.de.
authors.forename: J
authors.initials: J
authors.lastname: Gromoll
mesh_terms: D020413:3' Untranslated Regions; D000328:Adult; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D020641:Polymorphism, Single Nucleotide; D034622:RNA Interference; D011974:Receptors, LH / Q000235:genetics* / Q000378:metabolism; D027724:Reproductive Techniques, Assisted
chemical_list: D020413:3' Untranslated Regions; C546275:MIRN513A1 microRNA, human; D035683:MicroRNAs; D011974:Receptors, LH
keywords: Chorionic gonadotropin receptor (LHCGR); Granulosa cells; Luteinizing hormone; Post-transcriptional regulation; Reproduction; microRNA