Molecular Analysis of a Mutated FSH Receptor Detected in a Patient with Spontaneous Ovarian Hyperstimulation Syndrome

Sayaka Uchida, Hiroshi Uchida, Tetsuo Maruyama, Takashi Kajitani, Hideyuki Oda, Kaoru Miyazaki, Maki Kagami, Yasunori Yoshimura

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Abstract

Spontaneous ovarian hyperstimulation syndrome (sOHSS) is a rare event that may result from a FSH-producing pituitary adenoma (FSHoma), activating mutations of the FSH receptor (FSHR), and cross-reactivity of the FSHR to elevated hCG and TSH in the setting of pregnancy or hypothyroidism. The objective of this study was to investigate whether an aberrant FSHR was present in a woman with sOHSS and a non-surgically diagnosed FSHoma whose serum FSH levels and FSH bioactivity were nearly normal. Sequencing of the patient's FSHR gene revealed a heterozygous novel missense mutation c. 1536G>A resulting in an amino acid substitution M512I. We asked whether this mutant FSHR affected FSHR-mediated signaling pathways involving cAMP/protein kinase A (PKA), phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) and v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog kinase (SRC)/ p42/p44 extracellular signal-regulated protein kinases (ERK1/2). Thus, 293T cells expressing wild-type (FSHRwt), the mutant FSHR (FSHRmt), or both (FSHRwt/mt) were treated with FSH and subjected to measurements of intracellular cAMP, cAMP-induced CRE (cAMP response element)-mediated luciferase assays and immunoblot analyses of phosphorylated PI3K and ERK1/2. There were no differences in luciferase activities or phosphorylation levels of ERK1/2 among FSHRwt, FSHRmt cells and FSHwt/mt cells. However, FSHRmt cells showed a significant reduction in both cAMP production and PI3K phosphorylation levels with unchanged phosphorylation of ERK1/2 upon FSH stimulation in comparison to FSHwt cells. Also, FSH treatment did not provoke PI3K phosphorylation in FSHwt/mt cells. These results indicate that the novel missense M512I FSHR mutation identified herein did not participate in hyperactivation of FSHR-mediated signaling pathways but rather in hypoactivation of the FSH-mediated PI3K/AKT pathway. Thus, this study demonstrates a new functional property of this novel mutatnt FSHR, which, however, might not be involved in the pathogenesis of sOHSS in this FSHoma patient.

Original languageEnglish
Article numbere75478
JournalPLoS One
Volume8
Issue number9
DOIs
Publication statusPublished - 2013 Sep 13

Fingerprint

FSH Receptors
Ovarian Hyperstimulation Syndrome
Phosphatidylinositol 3-Kinase
receptors
phosphatidylinositol 3-kinase
Phosphorylation
phosphorylation
mutants
cells
luciferase
Luciferases
protein kinases
mutation
Proto-Oncogene Proteins c-akt
Mutation
missense mutation
hypothyroidism
cAMP-dependent protein kinase
HEK293 Cells
response elements

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Molecular Analysis of a Mutated FSH Receptor Detected in a Patient with Spontaneous Ovarian Hyperstimulation Syndrome. / Uchida, Sayaka; Uchida, Hiroshi; Maruyama, Tetsuo; Kajitani, Takashi; Oda, Hideyuki; Miyazaki, Kaoru; Kagami, Maki; Yoshimura, Yasunori.

In: PLoS One, Vol. 8, No. 9, e75478, 13.09.2013.

Research output: Contribution to journalArticle

Uchida, Sayaka ; Uchida, Hiroshi ; Maruyama, Tetsuo ; Kajitani, Takashi ; Oda, Hideyuki ; Miyazaki, Kaoru ; Kagami, Maki ; Yoshimura, Yasunori. / Molecular Analysis of a Mutated FSH Receptor Detected in a Patient with Spontaneous Ovarian Hyperstimulation Syndrome. In: PLoS One. 2013 ; Vol. 8, No. 9.
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