title: Variance in total levels of phospholipase C zeta (PLC-ζ) in human sperm may limit the applicability of quantitative immunofluorescent analysis as a diagnostic indicator of oocyte activation capability.
abstract: OBJECTIVE
  To examine whether similar levels of phospholipase C zeta (PLC-ζ) protein are present in sperm from men whose ejaculates resulted in normal oocyte activation, and to examine whether a predominant pattern of PLC-ζ localization is linked to normal oocyte activation ability.
  
  
  METHODS
  Laboratory study.
  
  
  METHODS
  University laboratory.
  
  
  METHODS
  Control subjects (men with proven oocyte activation capacity; n = 16) and men whose sperm resulted in recurrent intracytoplasmic sperm injection failure (oocyte activation deficient [OAD]; n = 5).
  
  
  METHODS
  Quantitative immunofluorescent analysis of PLC-ζ protein in human sperm.
  
  
  METHODS
  Total levels of PLC-ζ fluorescence, proportions of sperm exhibiting PLC-ζ immunoreactivity, and proportions of PLC-ζ localization patterns in sperm from control and OAD men.
  
  
  RESULTS
  Sperm from control subjects presented a significantly higher proportion of sperm exhibiting PLC-ζ immunofluorescence compared with infertile men diagnosed with OAD (82.6% and 27.4%, respectively). Total levels of PLC-ζ in sperm from individual control and OAD patients exhibited significant variance, with sperm from 10 out of 16 (62.5%) exhibiting levels similar to OAD samples. Predominant PLC-ζ localization patterns varied between control and OAD samples with no predictable or consistent pattern.
  
  
  CONCLUSIONS
  The results indicate that sperm from control men exhibited significant variance in total levels of PLC-ζ protein, as well as significant variance in the predominant localization pattern. Such variance may hinder the diagnostic application of quantitative PLC-ζ immunofluorescent analysis.
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: Junaid
authors.initials: J
authors.lastname: Kashir
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: Celine
authors.initials: C
authors.lastname: Jones
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: Ginny
authors.initials: G
authors.lastname: Mounce
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: Walaa M
authors.initials: WM
authors.lastname: Ramadan
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Lemmon
authors.affiliation: Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium.
authors.forename: Bjorn
authors.initials: B
authors.lastname: Heindryckx
authors.affiliation: Department for Reproductive Medicine, Ghent University Hospital, Ghent, Belgium.
authors.forename: Petra
authors.initials: P
authors.lastname: de Sutter
authors.affiliation: Department of Pharmacology, University of Oxford, Oxford, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Parrington
authors.affiliation: Oxford Fertility Unit, Institute of Reproductive Sciences, Oxford, United Kingdom.
authors.forename: Karen
authors.initials: K
authors.lastname: Turner
authors.affiliation: Oxford Fertility Unit, Institute of Reproductive Sciences, Oxford, United Kingdom.
authors.forename: Tim
authors.initials: T
authors.lastname: Child
authors.affiliation: Oxford Fertility Unit, Institute of Reproductive Sciences, Oxford, United Kingdom.
authors.forename: Enda
authors.initials: E
authors.lastname: McVeigh
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Women's Centre, John Radcliffe Hospital, Headington, Oxford, United Kingdom. Electronic address: kevin.coward@obs-gyn.ox.ac.uk.
authors.forename: Kevin
authors.initials: K
authors.lastname: Coward
mesh_terms: D000328:Adult; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis* / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D054801:Phosphoinositide Phospholipase C / Q000378:metabolism*; D027724:Reproductive Techniques, Assisted; D020554:Sperm Injections, Intracytoplasmic; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology*
chemical_list: C499235:PLCZ1 protein, human; D054801:Phosphoinositide Phospholipase C