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.)
- EMBRYO DEVELOPMENT
- cross correlation
- PARTHENOGENETIC ACTIVATION
- phospholipase C zeta
- MOUSE EGGS
- particle image velocimetry
- EGG ACTIVATION