Abstract
BackgroundHatching is crucial for mammalian embryo implantation, since difficulties during this process can lead to implantation failure, ectopic pregnancy and consequent infertility. Despite years of intensive researches, how internal and external factors affecting embryo hatch are still largely unclear.MethodsThe effects of parental genetic material and oxygen concentration on hatch process were examined. Fertilized and parthenogenetic mouse preimplantation embryos were cultured in vitro under 5 and 20% oxygen for 120 h. Zona pellucida drilling by Peizo micromanipulation were performed to resemble the breach by sperm penetration.ResultsFirstly, parthenogenetic embryos had similarly high blastocyst developmental efficiency as fertilized embryos, but significantly higher hatch ratio than fertilized embryos in both O2 concentrations. 5% O2 reduced the hatch rate of fertilized embryos from 58.2 to 23.8%, but increased that of parthenogenetic embryos from 81.2 to 90.8% significantly. Analogously, 5% O2 decreased the ratio of Oct4-positive cells in fertilized blastocysts, whereas increased that in parthenogenetic blastocysts. Additionally, 5% O2 increased the total embryonic cell number in both fertilized and parthegenetic embryos, when compared to 20% O2, and the total cell number of fertilized embryos was also higher than that of parthegenetic embryos, despite O2 concentration. Real-time PCR revealed that the expression of key genes involving in MAPK pathway and superoxide dismutase family might contribute to preimplantation development and consequent blastocyst hatch in vitro. Finally, we showed that fertilized and parthenogenetic embryos have diverse hatch dynamics in vitro, although the zona pellucida integrity is not the main reason for their mechanistic differences.ConclusionBoth parental genetic material and O2 concentration, as the representative of intrinsic and extrinsic factors respectively, have significant impacts on mouse preimplantation development and subsequent hatch dynamics, probably by regulating the gene expression involving in MAPK pathway and superoxide dismutase family to control embryonic cell proliferation and allocation of ICM cells.
Highlights
Hatching is crucial for mammalian embryo implantation, since difficulties during this process can lead to implantation failure, ectopic pregnancy and consequent infertility
Hatching is a process essential for mammalian embryo implantation, in which the embryo at blastocyst stage escapes from its zona pellucida (ZP), an outer shell which is composed of glycoproteins and is responsible for the preventation of polyspermy and ectopic pregnancy [1,2,3]
Significant difference was defined as P < 0.05. Both the oxygen concentration and genetic material affect the blastocyst hatch process in vitro To assess the effects of oxygen concentration and genetic material on embryo development and hatch process, we carried out four combinatorial treatments in a twofactor design by using the in vitro model of embryo culture, in which both fertilized zygotes from naturally mated female mice and parthenogenetic embryos from chemically activated MII oocytes were cultured for 120 h in 5 and 20% O2 respectively
Summary
Hatching is crucial for mammalian embryo implantation, since difficulties during this process can lead to implantation failure, ectopic pregnancy and consequent infertility. Other studies argued that there are no beneficial effects of 5% O2 culture on embryo subsequent development, such as implantation rate, pregnancy rate and fetal weight, when compared with atmospheric O2 culture [19, 26, 27]. It is still controversial whether low O2 concentrations can be adopted as the standard for human embryo culture, at least for blastocyst formation. One of aims in our current study is to find out the relationship between O2 concentration and embryo hatching ability in vitro
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