title: Follicle-stimulating hormone receptor (FSHR) alternative skipping of exon 2 or 3 affects ovarian response to FSH.
abstract: Genes critical for fertility are highly conserved in mammals. Interspecies DNA sequence variation, resulting in amino acid substitutions and post-transcriptional modifications, including alternative splicing, are a result of evolution and speciation. The mammalian follicle-stimulating hormone receptor (FSHR) gene encodes distinct species-specific forms by alternative splicing. Skipping of exon 2 of the human FSHR was reported in women of North American origin and correlated with low response to ovarian stimulation with exogenous follicle-stimulating hormone (FSH). To determine whether this variant correlated with low response in women of different genetic backgrounds, we performed a blinded retrospective observational study in a Turkish cohort. Ovarian response was determined as low, intermediate or high according to retrieved oocyte numbers after classifying patients in four age groups (<35, 35-37, 38-40, >40). Cumulus cells collected from 96 women undergoing IVF/ICSI following controlled ovarian hyperstimulation revealed four alternatively spliced FSHR products in seven patients (8%): exon 2 deletion in four patients; exon 3 and exons 2 + 3 deletion in one patient each, and a retention of an intron 1 fragment in one patient. In all others (92%) splicing was intact. Alternative skipping of exons 2, 3 or 2 + 3 were exclusive to low responders and was independent of the use of agonist or antagonist. Interestingly, skipping of exon 3 occurs naturally in the ovaries of domestic cats--a good comparative model for human fertility. We tested the signaling potential of human and cat variants after transfection in HEK293 cells and FSH stimulation. None of the splicing variants initiated cAMP signaling despite high FSH doses, unlike full-length proteins. These data substantiate the occurrence of FSHR exon skipping in a subgroup of low responders and suggest that species-specific regulation of FSHR splicing plays diverse roles in mammalian ovarian function.
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Cengiz
authors.initials: C
authors.lastname: Karakaya
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA Present address: Department of Obstetrics and Gynecology, Ohio State University College of Medicine, Columbus, OH 43210, USA.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Guzeloglu-Kayisli
authors.affiliation: Smithsonian Conservation Biology Institute, National Zoological Park, Washington, DC 20008, USA Present address: Taronga Conservation Society Australia, Taronga Western Plains Zoo, Duboo, NSW 2830, Australia.
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Hobbs
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Tsilya
authors.initials: T
authors.lastname: Gerasimova
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Asli
authors.initials: A
authors.lastname: Uyar
authors.affiliation: Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Erdem
authors.affiliation: Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Mesut
authors.initials: M
authors.lastname: Oktem
authors.affiliation: Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Erdem
authors.affiliation: Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Seyhan
authors.initials: S
authors.lastname: Gumuslu
authors.affiliation: Division of Reproductive Endocrinology and Infertility, IVF Center, Department of Obstetrics and Gynecology, Gazi University Medical School, Ankara 06500 Turkey.
authors.forename: Deniz
authors.initials: D
authors.lastname: Ercan
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA Present address: Boston IVF, Waltham, MA 02451, USA.
authors.forename: Denny
authors.initials: D
authors.lastname: Sakkas
authors.affiliation: Smithsonian Conservation Biology Institute, National Zoological Park, Washington, DC 20008, USA.
authors.forename: Pierre
authors.initials: P
authors.lastname: Comizzoli
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA.
authors.forename: Emre
authors.initials: E
authors.lastname: Seli
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT 06510, USA maria.lalioti@yale.edu.
authors.forename: Maria D
authors.initials: MD
authors.lastname: Lalioti
mesh_terms: D000328:Adult; D017398:Alternative Splicing; D000818:Animals; D002415:Cats; D054885:Cumulus Cells / Q000187:drug effects / Q000378:metabolism; D005091:Exons; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D007247:Infertility, Female / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D010053:Ovary / Q000187:drug effects / Q000378:metabolism*; D010062:Ovulation Induction; D011962:Receptors, FSH / Q000235:genetics / Q000378:metabolism*; D012189:Retrospective Studies
chemical_list: D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone
keywords: FSHR; IVF or ICSI; alternative splicing; cumulus cells; ovarian response