title: Characterization of human sperm populations using conventional parameters, surface ubiquitination, and apoptotic markers.
abstract: OBJECTIVE
  To directly compare distinct assays proposed to monitor human sperm quality and possibly preselect sperm populations for assisted reproductive technology (ART).
  
  
  METHODS
  Analysis of human sperm sample quality using several methodologies.
  
  
  METHODS
  Academic and clinical institutions.
  
  
  METHODS
  Samples from consenting patients undergoing routine semen analysis or ART.
  
  
  METHODS
  Human sperm samples were analyzed in terms of World Health Organization parameters and processed for annexin V, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling of DNA (TUNEL), and the sperm-ubiquitin tag immunoassay (SUTI). Samples were analyzed both by flow cytometry and fluorescence microscopy.
  
  
  METHODS
  Correlations among apoptotic markers (outer leaflet phosphatidylserine exposure, membrane integrity, and DNA fragmentation), external ubiquitination, and semen parameters in human spermatozoa.
  
  
  RESULTS
  Nonviable sperm, TUNEL-positive cells, and ubiquitin fluorescence intensity means inversely correlate with semen parameters. Apoptotic markers do not correlate with sperm surface ubiquitination. Normozoospermic samples have a higher number of viable cells and lower DNA fragmentation compared with samples with abnormal parameters. Nonviable sperm are more prevalent in samples with low counts and poor morphology but not low motility. Not all sperm with morphologic abnormalities present surface ubiquitination.
  
  
  CONCLUSIONS
  Sperm quality is inversely correlated with lack of viability, DNA fragmentation, and ubiquitin fluorescence intensity means. However, none of the apoptotic markers correlate with ubiquitin labeling. Elimination of defective sperm cells prior to ART using surface markers (annexin V, ubiquitin) seems unwarranted at this stage.
authors.affiliation: Center for Neuroscience and Cell Biology, Department of Zoology, University of Coimbra, Coimbra, Portugal.
authors.forename: Sandra
authors.initials: S
authors.lastname: Varum
authors.forename: Carla
authors.initials: C
authors.lastname: Bento
authors.forename: Ana Paula M
authors.initials: AP
authors.lastname: Sousa
authors.forename: Carina S S
authors.initials: CS
authors.lastname: Gomes-Santos
authors.forename: Paula
authors.initials: P
authors.lastname: Henriques
authors.forename: Teresa
authors.initials: T
authors.lastname: Almeida-Santos
authors.forename: Cristina
authors.initials: C
authors.lastname: Teodósio
authors.forename: Artur
authors.initials: A
authors.lastname: Paiva
authors.forename: João
authors.initials: J
authors.lastname: Ramalho-Santos
mesh_terms: D017304:Annexin A5 / Q000032:analysis; D017209:Apoptosis / Q000502:physiology*; D002470:Cell Survival; D053938:DNA Fragmentation; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D008856:Microscopy, Fluorescence; D010718:Phosphatidylserines / Q000378:metabolism; D011744:Pyrimidinones; D013081:Sperm Motility; D013094:Spermatozoa / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D025801:Ubiquitin / Q000032:analysis
chemical_list: D017304:Annexin A5; D010718:Phosphatidylserines; D011744:Pyrimidinones; D025801:Ubiquitin; C003954:merocyanine dye