Phospholipase C-ζ-induced Ca^(2+) oscillations cause coincident cytoplasmic movements in human oocytes that failed to fertilize after intracytoplasmic sperm injection

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.

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