title: Detailed functional studies on androgen receptor mild mutations demonstrate their association with male infertility.
abstract: BACKGROUND
  Mutations in the androgen receptor (AR) gene can cause the androgen insensitivity syndrome (AIS). For complete and severe partial AIS, well-characterized in vitro functional assays can be used for genotype-phenotype correlation; however, for mild forms of AIS, as associated with male infertility, experimental evidence is scarce or lacking. In particular, optimal in vitro functional tests informative about the genotype-phenotype relation have not been described.
  
  
  OBJECTIVE
  The objective of this study was to investigate the association among genotype and phenotype for AR mutations found in infertile males by conventional functional assays and additional in-depth studies performed with several gene reporters.
  
  
  METHODS
  To this aim, we selected four AR missense mutations associated with isolated male infertility (L547F and two novel mutations A474V and S650G) or partial AIS (Y571H). After introduction of the specific mutations in AR expression plasmid, we performed classical in vitro studies (Western immunoblotting, electrophoretic mobility shift assay, hormone-response curves) and transactivation assays with different reporter constructs (MMTV, Sc-ARU-TK, TAT-GRE- 2X, Slp-ARU-TK and PEM).
  
  
  CONCLUSIONS
  Our results showed that standard functional tests provide sufficient information only for severe AR mutations, whereas for AR mutations found in mild AIS patients with male infertility, only an extensive analysis with different in vitro systems, and in particular with PEM promoter, can give information on the functionality of the AR and therefore on the pathogenicity of the mutations and on genotype-phenotype correlation.
authors.affiliation: Department of Histology, Microbiology and Medical Biotechnology, University of Padova, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Zuccarello
authors.forename: A
authors.initials: A
authors.lastname: Ferlin
authors.forename: C
authors.initials: C
authors.lastname: Vinanzi
authors.forename: E
authors.initials: E
authors.lastname: Prana
authors.forename: A
authors.initials: A
authors.lastname: Garolla
authors.forename: L
authors.initials: L
authors.lastname: Callewaert
authors.forename: F
authors.initials: F
authors.lastname: Claessens
authors.forename: A O
authors.initials: AO
authors.lastname: Brinkmann
authors.forename: C
authors.initials: C
authors.lastname: Foresta
mesh_terms: D000328:Adult; D013734:Androgen-Insensitivity Syndrome / Q000235:genetics*; D005838:Genotype; D006801:Humans; D066298:In Vitro Techniques; D007248:Infertility, Male / Q000235:genetics*; D008297:Male; D020125:Mutation, Missense; D010641:Phenotype; D010957:Plasmids; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism; D012720:Severity of Illness Index
chemical_list: D011944:Receptors, Androgen