title: Fluorescence in situ hybridization and molecular studies in infertile men with dysplasia of the fibrous sheath.
abstract: OBJECTIVE
  To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS).
  
  
  METHODS
  Retrospective study.
  
  
  METHODS
  Regional Referral Center for Male Infertility, Siena, Italy.
  
  
  METHODS
  Twelve infertile patients with DFS sperm defects.
  
  
  METHODS
  Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis.
  
  
  METHODS
  The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction.
  
  
  RESULTS
  The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls.
  
  
  CONCLUSIONS
  These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.
authors: Department of Pediatrics, Obstetrics and Reproductive Medicine, Siena University, Regional Referral Center for Male Infertility, Azienda Ospedaliera Universitaria Senese, Siena, Italy. baccetti@unisi.it
authors: Baccio
authors: B
authors: Baccetti
authors: Giulia
authors: G
authors: Collodel
authors: Laura
authors: L
authors: Gambera
authors: Elena
authors: E
authors: Moretti
authors: Francesca
authors: F
authors: Serafini
authors: Paola
authors: P
authors: Piomboni
mesh_terms: D000328:Adult; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D041321:Chromosomes, Human, X / Q000235:genetics; D041322:Chromosomes, Human, Y / Q000235:genetics; D004171:Diploidy; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007248:Infertility, Male / Q000235:genetics* / Q000473:pathology*; D008297:Male; D012189:Retrospective Studies; D013094:Spermatozoa / Q000473:pathology* / Q000502:physiology* / Q000648:ultrastructure