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Fertil Steril. 2012 Mar;97(3):742-7. doi: 10.1016/j.fertnstert.2011.12.013. Epub 2012 Jan 2.

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

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  • 1School of Medicine, Cardiff University, Heath Park, Cardiff, United Kingdom. Swannk1@cardiff.ac.uk

Abstract

OBJECTIVE:

To evaluate the imaging of cytoplasmic movements in human oocytes as a potential method to monitor the pattern of Ca(2+) 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 (ζ) cRNA and a Ca(2+)-sensitive fluorescent dye.

MAIN OUTCOME MEASURE(S):

Simultaneous detection of oocyte cytoplasmic movements using particle image velocimetry (PIV) and of Ca(2+) oscillations using a Ca(2+)-sensitive fluorescent dye.

RESULT(S):

Microinjection of PLCζ cRNA into human oocytes that had failed to fertilize after ICSI resulted in the appearance of prolonged Ca(2+) oscillations. Each transient Ca(2+) 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 Ca(2+) 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 Ca(2+) oscillations in human zygotes.

Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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