title: Assessment of acrosome and nuclear abnormalities in human spermatozoa with large vacuoles.
abstract: BACKGROUND
  Spermatozoa with large vacuoles (SLV) may have a negative impact on embryo development. The origin of these vacuoles is unknown. We evaluated acrosome and nucleus alterations in isolated SLV, versus unselected spermatozoa.
  
  
  METHODS
  We studied 20 patients with teratozoospermia. Spermatozoa from the native semen sample and spermatozoa presenting a vacuole occupying >13.0% total head area, isolated under high magnification (×6600), were assessed. Confocal and transmission electron microscope evaluations were performed on SLV and native sperm, respectively. Acrosome morphology and DNA fragmentation were analysed using proacrosin immunolabelling (monoclonal antibody 4D4) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay. Chromatin condensation was evaluated with aniline blue staining. Sperm aneuploidy was assessed using fluorescence in situ hybridization.
  
  
  RESULTS
  SLV represented 38.0 ± 5.10% of motile spermatozoa obtained after gradient density centrifugation. Vacuoles were mainly in the anterior and median sperm head (45.7 ± 2.90 and 46.1 ± 3.00%, respectively). Abnormal acrosomes were increased in SLV compared with unselected spermatozoa (77.8 ± 2.49 versus 70.6 ± 2.62%; P = 0.014). Microscopic observations showed an exclusively nuclear localization of large vacuoles. Complete DNA fragmentation was higher in native spermatozoa (P < 0.0001) than SLV, while chromatin condensation was altered in SLV (P < 0.0001). Aneuploidy and diploidy rates were increased in SLV (P < 0.0001).
  
  
  CONCLUSIONS
  Sperm vacuoles were exclusively nuclear. In our selected teratozoospermic population, aneuploidy and chromatin condensation defects were the main alterations observed in SLV. Based on results from this small sample of spermatozoa, we propose a global impairment of the spermatogenesis process as a common origin of the morphological alterations.
authors.affiliation: EA 4308 Spermatogenesis and Male Gamete Quality, Reproductive Biology Laboratory-CECOS, Rouen University Hospital, University of Rouen, Rouen, France.
authors.forename: A
authors.initials: A
authors.lastname: Perdrix
authors.forename: A
authors.initials: A
authors.lastname: Travers
authors.forename: M H
authors.initials: MH
authors.lastname: Chelli
authors.forename: D
authors.initials: D
authors.lastname: Escalier
authors.forename: J L
authors.initials: JL
authors.lastname: Do Rego
authors.forename: J P
authors.initials: JP
authors.lastname: Milazzo
authors.forename: N
authors.initials: N
authors.lastname: Mousset-Siméon
authors.forename: B
authors.initials: B
authors.lastname: Macé
authors.forename: N
authors.initials: N
authors.lastname: Rives
mesh_terms: D000177:Acrosome / Q000648:ultrastructure*; D000328:Adult; D000782:Aneuploidy; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D053938:DNA Fragmentation; D047108:Embryonic Development; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D020287:In Situ Nick-End Labeling; D007248:Infertility, Male / Q000473:pathology*; D008297:Male; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D055101:Semen Analysis / Q000379:methods*; D013094:Spermatozoa / Q000648:ultrastructure*; D014617:Vacuoles / Q000648:ultrastructure*
chemical_list: D002843:Chromatin