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

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• 作者：Karl Swann ; Ph.D.^a ; ^{Swannk1@cardiff.ac.uk} ; Shane Windsor ; Ph.D.^b ; Karen Campbell ; Ph.D.^c ; Khalil Elgmati ; M.Sc.^a ; Michail Nomikos ; Ph.D.^a ; Magdalena Zernicka-Goetz ; Ph.D.^d ; Nazar Amso ; Ph.D.^a ; F. Anthony Lai ; Ph.D.^a ; Adrian Thomas ; Ph.D.^b ; Christopher Graham ; D.Phil.^b
• 关键词：Oocyte ; zygote ; calcium ; movement ; phospholipase C zeta ; cross correlation ; particle image velocimetry
• 刊名：Fertility and Sterility
• 出版年：2012
• 出版时间：March, 2012
• 年：2012
• 卷：97
• 期：3
• 页码：742-747
• 全文大小：315 K

文摘

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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 (¦Æ) 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¦Æ 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.