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Hum. Reprod. Advance Access originally published online on November 14, 2007
Human Reproduction 2008 23(2):365-373; doi:10.1093/humrep/dem350
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Preimplantation development of mouse oocytes activated by different levels of human phospholipase C zeta

Y. Yu1, C.M. Saunders2, F.A. Lai2 and K. Swann1,3

1 Department of Obstetrics and Gynaecology, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK 2 Wales Heart Research Institute, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK

3 Correspondence address. Tel: +44 2920 742039; Fax: +44 2920 744399; E-mail: swannk1{at}cardiff.ac.uk

BACKGROUND: A sperm-specific phospholipase C zeta (PLC{zeta}) has been shown to trigger Ca2+ oscillations in mouse and human oocytes and appears to be the sperm factor responsible for activation at fertilization. Previously, complementary RNA (cRNA) injection was used to introduce PLC{zeta} into oocytes, but it was unclear how much PLC{zeta} protein is required for development. Here we have injected cRNA encoding luciferase-tagged human PLC{zeta} (hPLC{zeta}-luc) into mouse oocytes and established the relationship between hPLC{zeta}-luc expression, Ca2+ oscillations and development.

METHODS: Mouse oocytes were injected with hPLC{zeta}-luc cRNA and a fluorescent Ca2+dye to monitor hPLC{zeta}-luc expression and Ca2+ oscillations, respectively. After inducing diploidy, development in vitro was monitored in hPLC{zeta}-luc cRNA microinjected oocytes and compared with parallel oocytes activated by incubation in Sr2+.

RESULTS: Repetitive Ca2+ oscillations and oocyte activation were triggered by hPLC{zeta} over a wide range of luciferase expression levels. However, subsequent development of embryos to the blastocyst stage was observed only when expression of hPLC{zeta}-luc was optimized within a specific range. The blastocyst cell number was also affected by the level of hPLC{zeta} expression.

CONCLUSIONS: Human PLC{zeta} can readily activate mouse oocytes, however, effective development to blastocyst stages is only achieved within a specific window of hPLC{zeta}-luc protein expression levels.

Key words: calcium/fertilization/oocyte/activation/human phospholipase C zeta

Submitted on June 14, 2007; resubmitted on July 24, 2007; accepted on July 28, 2007.


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