亲代遗传物质和氧浓度影响体外培养的小鼠胚胎孵化动力学。
Parental genetic material and oxygen concentration affect hatch dynamics of mouse embryo in vitro.
机构信息
Center for Reproductive Medicine, Key Laboratory for Reproductive Medicine of Guangdong Province, Key Laboratory for Major Obstetric Diseases of Guangdong Province, and Key Laboratory for Reproduction and Genetics of Guangdong Higher Education Institutes, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China.
Beijing Acorndx Biotechnology Co. Ltd, Beijing, 100176, China.
出版信息
Reprod Biol Endocrinol. 2018 Apr 21;16(1):39. doi: 10.1186/s12958-018-0356-8.
BACKGROUND
Hatching 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.
METHODS
The 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.
RESULTS
Firstly, parthenogenetic embryos had similarly high blastocyst developmental efficiency as fertilized embryos, but significantly higher hatch ratio than fertilized embryos in both O concentrations. 5% O 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% O decreased the ratio of Oct4-positive cells in fertilized blastocysts, whereas increased that in parthenogenetic blastocysts. Additionally, 5% O increased the total embryonic cell number in both fertilized and parthegenetic embryos, when compared to 20% O, and the total cell number of fertilized embryos was also higher than that of parthegenetic embryos, despite O 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.
CONCLUSION
Both parental genetic material and O 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.
背景
孵化对于哺乳动物胚胎着床至关重要，因为这一过程中的困难可能导致着床失败、宫外孕和随之而来的不孕。尽管经过多年的深入研究，内部和外部因素如何影响胚胎孵化仍在很大程度上不清楚。
方法
研究了亲代遗传物质和氧浓度对孵化过程的影响。将受精和孤雌激活的小鼠胚胎在体外分别于 5%和 20%的氧浓度下培养 120 小时。采用压电微操作进行透明带钻孔，模拟精子穿透时的破裂。
结果
首先，孤雌胚胎的囊胚发育效率与受精胚胎相似，但在两种氧浓度下的孵化率均显著高于受精胚胎。5%的氧降低了受精胚胎的孵化率，从 58.2%降至 23.8%，但显著增加了孤雌胚胎的孵化率，从 81.2%增至 90.8%。类似地，5%的氧降低了受精囊胚中 Oct4 阳性细胞的比例，而增加了孤雌囊胚中的比例。此外，与 20%的氧相比，5%的氧增加了受精和孤雌胚胎的总胚胎细胞数量，并且受精胚胎的总细胞数量也高于孤雌胚胎，尽管氧浓度不同。实时 PCR 显示，涉及 MAPK 途径和超氧化物歧化酶家族的关键基因的表达可能有助于体外胚胎的植入前发育和随后的囊胚孵化。最后，我们表明，尽管透明带完整性不是它们机制差异的主要原因，但受精和孤雌胚胎在体外具有不同的孵化动力学。
结论
亲代遗传物质和氧浓度（分别代表内在和外在因素）都对小鼠植入前发育和随后的孵化动力学有显著影响，可能通过调节涉及 MAPK 途径和超氧化物歧化酶家族的基因表达来控制胚胎细胞的增殖和 ICM 细胞的分配。
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