title: Phospholipase Cζ rescues failed oocyte activation in a prototype of male factor infertility.
abstract: OBJECTIVE
  To determine the effect of infertility-linked sperm phospholipase Cζ (PLCζ) mutations on their ability to trigger oocyte Ca(2+) oscillations and development, and also to evaluate the potential therapeutic utility of wild-type, recombinant PLCζ protein for rescuing failed oocyte activation and embryo development.
  
  
  METHODS
  Test of a novel therapeutic approach to male factor infertility.
  
  
  METHODS
  University medical school research laboratory.
  
  
  METHODS
  Donated unfertilized human oocytes from follicle reduction.
  
  
  METHODS
  Microinjection of oocytes with recombinant human PLCζ protein or PLCζ cRNA and a Ca(2+)-sensitive fluorescent dye.
  
  
  METHODS
  Measurement of the efficacy of mutant and wild-type PLCζ-mediated enzyme activity, oocyte Ca(2+) oscillations, activation, and early embryo development.
  
  
  RESULTS
  In contrast to the wild-type protein, mutant forms of human sperm PLCζ display aberrant enzyme activity and a total failure to activate unfertilized oocytes. Subsequent microinjection of recombinant human PLCζ protein reliably triggers the characteristic pattern of cytoplasmic Ca(2+) oscillations at fertilization, which are required for normal oocyte activation and successful embryo development to the blastocyst stage.
  
  
  CONCLUSIONS
  Dysfunctional sperm PLCζ cannot trigger oocyte activation and results in male factor infertility, so a potential therapeutic approach is oocyte microinjection of active, wild-type PLCζ protein. We have demonstrated that recombinant human PLCζ can phenotypically rescue failed activation in oocytes that express dysfunctional PLCζ, and that this intervention culminates in efficient blastocyst formation.
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom; National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece. Electronic address: mixosn@yahoo.com.
authors.forename: Michail
authors.initials: M
authors.lastname: Nomikos
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Yuansong
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Khalil
authors.initials: K
authors.lastname: Elgmati
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom; National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Theodoridou
authors.affiliation: IVF Wales, University Hospital of Wales, Heath Park, Cardiff, United Kingdom.
authors.forename: Karen
authors.initials: K
authors.lastname: Campbell
authors.affiliation: National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece.
authors.forename: Vyronia
authors.initials: V
authors.lastname: Vassilakopoulou
authors.affiliation: National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece.
authors.forename: Christos
authors.initials: C
authors.lastname: Zikos
authors.affiliation: National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece.
authors.forename: Evangelia
authors.initials: E
authors.lastname: Livaniou
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Nazar
authors.initials: N
authors.lastname: Amso
authors.affiliation: National Center for Scientific Research "Demokritos," Aghia Paraskevi, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Nounesis
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Karl
authors.initials: K
authors.lastname: Swann
authors.affiliation: Cardiff University School of Medicine, Heath Park, Cardiff, United Kingdom. Electronic address: lait@cf.ac.uk.
authors.forename: F Anthony
authors.initials: FA
authors.lastname: Lai
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007248:Infertility, Male / Q000188:drug therapy* / Q000503:physiopathology; D008297:Male; D051379:Mice; D008845:Microinjections; D023421:Models, Animal; D009154:Mutation / Q000235:genetics; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D013084:Sperm-Ovum Interactions / Q000187:drug effects* / Q000502:physiology
chemical_list: D011994:Recombinant Proteins; C499235:PLCZ1 protein, human; D054801:Phosphoinositide Phospholipase C; D002118:Calcium