title: A novel mutation (N233K) in the transactivating domain and the N756S mutation in the ligand binding domain of the androgen receptor gene are associated with male infertility.
abstract: OBJECTIVE
  Resistance to androgens has been suggested as a possible cause of male infertility. This hypothesis is based mainly on binding studies in genital skin fibroblasts but the molecular evidence is sparse.
  
  
  METHODS
  Molecular studies of the androgen receptor gene were performed in 10 azoo- or oligozoospermic men, presenting with clinical signs of low androgen activity-poor virilization and high serum LH despite elevated testosterone levels, but without genital malformations.
  
  
  METHODS
  Ten men with serum LH >10 IU/l and testosterone >30 nmol/l as well as a low sperm concentration < 20 x 106/ml.
  
  
  METHODS
  Genomic DNA was prepared from peripheral leucocytes and PCR-amplification of the coding region of androgen receptor was performed, followed by direct sequencing. Identified mutations were reconstructed by site-directed mutagenesis and the functional properties of the mutants were analysed, using transient expression in COS-1 cells and subsequent transactivation assays. Hormone binding assays were performed in genital skin fibroblasts from the patients.
  
  
  RESULTS
  Two of the 10 men were shown to have a mutation in the androgen receptor gene. Subject 1, who presented with azoospermia, serum testosterone (T) 50 nmol/l and LH 20 IU/l, had a mutation in exon 1, changing amino acid asparagine 233 to lysine (N233K). In fibroblasts cultured from genital skin, the receptor affinity for 5alpha-dihydrotestosterone (DHT) was normal as compared to healthy controls, but the receptor-hormone complex was thermolabile at 42 degrees C. Subject 2 exhibited severe oligozoospermia and a similar endocrine pattern (T = 50 nmol/l and LH = 25 IU/l). He had a mutation in exon 5 changing asparagine 756 to serine (N756S). The affinity for DHT in cultured genital fibroblasts from this patient was reduced. Transactivation was abnormal for both mutants, N233K reaching 46% and N756S 38% of wild type activity when stimulated with 10 nmol/l DHT.
  
  
  CONCLUSIONS
  Androgen receptor mutations may affect sperm production without resulting in genital malformations. Thus, in infertile men with a clinical presentation of poor androgen activity and an endocrine profile compatible with androgen resistance, mutations in the androgen receptor should be taken into consideration.
authors: Department of Molecular Medicine, Karolinska Hospital, Stockholm, Sweden. ay.giwercman@swipnet.se
authors: Y L
authors: YL
authors: Giwercman
authors: A
authors: A
authors: Nikoshkov
authors: B
authors: B
authors: Byström
authors: Pousette A
authors: S
authors: S
authors: Arver
authors: A
authors: A
authors: Wedell
mesh_terms: D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002478:Cells, Cultured; D013196:Dihydrotestosterone / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005640:Follicle Stimulating Hormone / Q000097:blood; D006801:Humans; D007248:Infertility, Male / Q000097:blood / Q000235:genetics* / Q000473:pathology; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011485:Protein Binding; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D012738:Sex Hormone-Binding Globulin / Q000032:analysis; D013076:Sperm Count; D013739:Testosterone / Q000097:blood; D014162:Transfection
chemical_list: D011944:Receptors, Androgen; D012738:Sex Hormone-Binding Globulin; D013196:Dihydrotestosterone; D013739:Testosterone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone