title: Motile sperm organelle morphology evaluation-selected globozoospermic human sperm with an acrosomal bud exhibits novel patterns and higher levels of phospholipase C zeta.
abstract: OBJECTIVE
  Does motile sperm organelle morphology examination (MSOME) affect levels and localization patterns of the oocyte activation factor phospholipase C zeta (PLCζ) in globozoospermic sperm with and without an acrosomal bud?
  
  
  CONCLUSIONS
  MSOME identified round-headed globozoospermic sperm with increased levels of PLCζ relative to sperm from the same sample that did not undergo MSOME, and identified novel patterns of PLCζ localization in sperm exhibiting an acrosomal bud.
  
  
  BACKGROUND
  Absence or reduction in the level of PLCζ in the sperm head, abnormal localization patterning, or defective functional ability as a result of PLCζ gene mutation, have been linked to certain types of human male factor infertility in which oocyte activation is deficient. It has been determined that a subpopulation of sperm (1%) from a patient exhibiting 100% globozoospermia presented with an acrosome bud upon MSOME. A cycle of intracytoplasmic morphologically selected sperm injection, carried out with sperm exhibiting an acrosomal bud led to pregnancy and birth of a healthy baby boy, without the use of assisted oocyte activation (AOA).
  
  
  METHODS
  Immunofluorescent analysis of PLCζ in globozoospermic sperm from three patients, before and after MSOME.
  
  
  METHODS
  Quantitative immunofluorescence was used to investigate PLCζ levels and localization patterns in individual sperm (n = 1 patient) identified by MSOME and isolated by micromanipulation, and presenting with and without the acrosomal bud. A secondary aim was to investigate levels and localization patterns of PLCζ in sperm before and after MSOME from two other globozoospermic men.
  
  
  RESULTS
  Non-globozoospermic control sperm exhibited characteristic localization patterns of PLCζ immunofluorescence. Completely round-headed globozoospermic sperm from patients 1-3 were either devoid of PLCζ immunofluorescence, or exhibited an abnormal, punctate, pattern of PLCζ localization. PLCζ immunofluorescence in sperm exhibiting an acrosomal bud was observed in the midpiece with varying fluorescent intensity and was detected in 28.5% of such sperm. The majority of sperm with an acrosomal bud (43.0%) exhibited punctate patterns of PLCζ localization within the sperm head. A further 28.5% of sperm exhibited PLCζ in both the head and the midpiece. Total levels of PLCζ, and the proportions of sperm exhibiting PLCζ immunoreactivity, showed significant variance (P ≤ 0.05) amongst control [45.8 arbitrary units (a.u.) and 95.7%, respectively], non-MSOME-selected (25.9 a.u. and 46.1%, respectively) and MSOME-selected globozoospermic sperm (33.4 a.u. and 65.0%, respectively). Total levels of PLCζ immunofluorescence, and proportions of sperm exhibiting PLCζ immunoreactivity, in control sperm was significantly higher (P≤ 0.05) compared with non-MSOME-selected sperm, but not significantly different from MSOME-selected sperm.
  
  
  CONCLUSIONS
  The low numbers of sperm analysed may not be ideal for conclusive statistical analysis. Evaluation of the effects of MSOME on morphologically normal sperm would confirm conclusions.
  
  
  CONCLUSIONS
  The present findings provide hope for the future treatment of globozoospermia without the need for AOA, and provide further evidence for the clinical application of PLCζ as a therapeutic and prognostic tool.
  
  
  BACKGROUND
  The research described herein was funded by the Nuffield Department of Obstetrics and Gynaecology, University of Oxford. The authors report no conflict of interest.
authors.affiliation: Nuffield Department of Obstetrics and Gynaecology, Level 3, Women's Centre,  John Radcliffe Hospital, Headington, Oxford OX3 9DU,  UK.
authors.forename: Junaid
authors.initials: J
authors.lastname: Kashir
authors.forename: Nathalie
authors.initials: N
authors.lastname: Sermondade
authors.forename: Christophe
authors.initials: C
authors.lastname: Sifer
authors.forename: Su Lin
authors.initials: SL
authors.lastname: Oo
authors.forename: Celine
authors.initials: C
authors.lastname: Jones
authors.forename: Ginny
authors.initials: G
authors.lastname: Mounce
authors.forename: Karen
authors.initials: K
authors.lastname: Turner
authors.forename: Tim
authors.initials: T
authors.lastname: Child
authors.forename: Enda
authors.initials: E
authors.lastname: McVeigh
authors.forename: Kevin
authors.initials: K
authors.lastname: Coward
mesh_terms: D000177:Acrosome / Q000378:metabolism / Q000473:pathology; D000328:Adult; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism / Q000473:pathology*; D008297:Male; D015388:Organelles / Q000378:metabolism / Q000473:pathology*; D054801:Phosphoinositide Phospholipase C / Q000378:metabolism*; D021381:Protein Transport; D055101:Semen Analysis / Q000009:adverse effects*; D013077:Sperm Head / Q000378:metabolism / Q000473:pathology*; D032961:Sperm Midpiece / Q000378:metabolism / Q000473:pathology
chemical_list: C499235:PLCZ1 protein, human; D054801:Phosphoinositide Phospholipase C