期刊
MOLECULAR HUMAN REPRODUCTION
卷 21, 期 10, 页码 783-791出版社
OXFORD UNIV PRESS
DOI: 10.1093/molehr/gav042
关键词
PLC zeta; ICSI; fertilization; sperm; oocyte
资金
- Libyan Ministry of Education
- EU-FP7 Marie Curie Intra-European Fellowship [628634]
- Cook Medical Technologies LLC.
- Cardiff University
Artificial oocyte activation to overcome failed fertilization after intracytoplasmic sperm injection (ICSI) in human oocytes typically employs Ca2+ ionophores to produce a single cytosolic Ca2+ increase. In contrast, recombinant phospholipase Czeta (PLC zeta) causes Ca2+ oscillations indistinguishable from those occurring during fertilization, but remains untested for its efficacy in a scenario of ICSI fertilization failure. Here, we compare PLC zeta with other activation stimuli in a mouse model of failed oocyte activation after ICSI, in which heat-treated sperm are injected into mouse oocytes. We show that increasing periods of 56 degrees C exposure of sperm produces a progressive loss of Ca2+ oscillations after ICSI. The decrease in Ca2+ oscillations produces a reduction in oocyte activation and embryo development to the blastocyst stage. We treated such oocytes that failed to activate after ICSI either with Ca2+ ionophore, or with Sr2+ media which causes Ca2+ oscillations, or we injected them with recombinant human PLC zeta. All these treatments rescued oocyte activation, although Sr2+ and PLC zeta gave the highest rates of development to blastocyst. When recombinant PLC zeta was given to oocytes previously injected with control sperm, they developed normally to the blastocyst stage at rates similar to that after control ICSI. The data suggest that recombinant human PLC zeta protein is an efficient means of rescuing oocyte activation after ICSI failure and that it can be effectively used even if the sperm already contains endogenous Ca2+ releasing activity.
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