title: Follicle-stimulating hormone receptor polymorphism (G-29A) is associated with altered level of receptor expression in Granulosa cells.
abstract: BACKGROUND
  Polymorphisms of the FSHR gene are associated with variable ovarian response to FSH stimulation in subjects undergoing in vitro fertilization (IVF) treatment. The type of ovarian response is correlated with the level of FSH receptor (FSHR) expression on granulosa cells.
  
  
  OBJECTIVE
  We investigated whether the polymorphism at position -29 in the promoter of the FSHR gene may contribute in altered receptor expression.
  
  
  METHODS
  FSHR polymorphism at position -29 was studied in 100 subjects undergoing IVF treatment. Association of this polymorphism with level of FSHR expression was retrospectively analyzed.
  
  
  METHODS
  The study was conducted at an academic research institute and private IVF clinic.
  
  
  METHODS
  The genotype at position -29 of the FSHR gene was studied in IVF subjects by PCR-restriction fragment length polymorphism. Total RNA and protein was extracted from granulosa cells. The relative FSHR mRNA expression was carried out by real-time PCR. The receptor protein expression was evaluated by Western blot and confocal microscopy.
  
  
  RESULTS
  The clinical and endocrinological parameters revealed that almost 72% of subjects with the AA genotype at position -29 of FSHR gene were poor ovarian responders (odds ratio 8.63, 95% confidential interval 1.84-45.79; P = 0.001). The lower cleavage intensity predicted by in silico analysis for A allele as compared with the G allele suggest the difference in the DNA-protein binding affinity. The relative expression of FSHR at mRNA and protein level was significantly reduced in subjects with AA genotype as compared with the GG genotype.
  
  
  CONCLUSIONS
  Poor ovarian response observed in subjects with the AA genotype at position -29 of the FSHR gene is due to reduced receptor expression.
authors.affiliation: National Institute for Research in Reproductive Health, Mumbai 400 012, India.
authors.forename: Swapna S
authors.initials: SS
authors.lastname: Desai
authors.forename: Swati K
authors.initials: SK
authors.lastname: Achrekar
authors.forename: Bhakti R
authors.initials: BR
authors.lastname: Pathak
authors.forename: Sadhana K
authors.initials: SK
authors.lastname: Desai
authors.forename: Vijay S
authors.initials: VS
authors.lastname: Mangoli
authors.forename: Ranjana V
authors.initials: RV
authors.lastname: Mangoli
authors.forename: Smita D
authors.initials: SD
authors.lastname: Mahale
mesh_terms: D000328:Adult; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology / Q000627:therapeutic use; D005838:Genotype; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D007247:Infertility, Female / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D010053:Ovary / Q000187:drug effects / Q000378:metabolism; D010062:Ovulation Induction; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D011962:Receptors, FSH / Q000235:genetics* / Q000378:metabolism
chemical_list: D011962:Receptors, FSH; D005640:Follicle Stimulating Hormone