Phospholipase C-zeta-induced Ca2+ oscillations cause coincident cytoplasmic movements in human oocytes that failed to fertilize after intracytoplasmic sperm injectiond

Karl Swann*, Shane Windsor, Karen Campbell, Khalil Elgmati, Michail Nomikos, Magdalena Zernicka-Goetz, Nazar Amso, Anthony Lai, Adrian Thomas, Christopher Graham

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

33 Citations (Scopus)

Abstract

Objective: To evaluate the imaging of cytoplasmic movements in human oocytes as a potential method to monitor the pattern of Ca2+ oscillations during activation.

Design: Test of a laboratory technique.

Setting: University medical school research laboratory.

Patient(s): Donated unfertilized human oocytes from intracytoplasmic sperm injection (ICSI) cycles.

Intervention(s): Microinjection of oocytes with phospholipase C (PLC) zeta (zeta) cRNA and a Ca2+-sensitive fluorescent dye.

Main Outcome Measure(s): Simultaneous detection of oocyte cytoplasmic movements using particle image velocimetry (PIV) and of Ca2+ oscillations using a Ca2+-sensitive fluorescent dye.

Result(s): Microinjection of PLC zeta cRNA into human oocytes that had failed to fertilize after ICSI resulted in the appearance of prolonged Ca2+ oscillations. Each transient Ca2+ concentration change was accompanied by a small coordinated movement of the cytoplasm that could be detected using PIV analysis.

Conclusion(s): The occurrence and frequency of cytoplasmic Ca2+ oscillations, a critical parameter in activating human zygotes, can be monitored by PIV analysis of cytoplasmic movements. This simple method provides a novel, noninvasive approach to determine in real time the occurrence and frequency of Ca2+ oscillations in human zygotes. (Fertil Steril (R) 2012; 97: 742-7. (C) 2012 by American Society for Reproductive Medicine.)

Original languageEnglish
Pages (from-to)742-747
Number of pages6
JournalFertility and Sterility
Volume97
Issue number3
DOIs
Publication statusPublished - Mar 2012

Keywords

  • calcium
  • zygote
  • CALCIUM
  • PROTEIN
  • Oocyte
  • EMBRYO DEVELOPMENT
  • cross correlation
  • IN-VITRO
  • PARTHENOGENETIC ACTIVATION
  • PLC-ZETA
  • HYDROLYSIS
  • movement
  • phospholipase C zeta
  • MOUSE EGGS
  • particle image velocimetry
  • EGG ACTIVATION

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