| Drinking alcohol during pregnancy significant ly increased the risk of fetal malformations.In recent years,the number of women who drink alcohol has increased and their age has decreased.Women drinking alcohol in an unknown pregnancy further increases the chances of alcohol exposure to the fetus,especially le ading to a high-risk environment for early(preimplantation)embryos.In previous work,we have found that alcohol exposure to early mouse embryos can lead to serious problems at all stages of embryo development,including pre-and post-implantation develop ment.At the same time,the tissue structure of placenta also showed obvious changes.As an important organ to ensure the normal development of the fetus,the placenta plays an irreplaceable function in many aspects,including the transport of nutrients an d the exchange of oxygen / carbon dioxide,as well as the release and regulation of reproductive endocrine hormones and the provision of physical barriers.Therefore,an in-depth understanding of the effect of alcohol exposure of early embryos on placental development and its related mechanism is an important link to fully understand the damage caused by alcohol to embryonic development.Studies have found that placental development is closely related to the expression of imprinted genes,and they are of great significance for placental development and functional exercise.As an important maternal imprinting gene,Plagl1(Pleiomorphic adenom a gene-like 1)plays an important role in the development of placenta and can affect the development of fetal adrenal gland,pituitary and fetal nervous system.In this study,E13.5 fetus and placenta were obtained by embryo culture in vitro and embryo tra nsfer.Enzyme-linked immunosorbent assay((ELISA)),real-time fluorescence quantitative PCR(RT-q PCR),immunohistochemist--ry(IHC)and sulfite sequencing(BSP)were used to explore the expression changes and potential regulatory mechanism of Plagl1 in placental development abnormalities induced by early embryonic alcohol exposure.The results showed that: 1)compared with the contro l group,alcohol exposure of early mouse embryos resulted in abnormal placental function,decreased expression of progesterone and estradiol,and changes in placental specific gene expression,glucose transporter expression and placental tissue at 13.5 day s.2)q PCR 、 Western Blotting and Immunohistochemistry were used to detect the expression of Plagl1 in early embryo blastocysts exposed to alcohol.The results showed that the expression of Plagl1 and Plakoglobin(γ-catenin,γ-Junction plakoglobin)in alcohol group was significantly higher than that in the control group.The expression of Plagl1 and Plakoglobin in the placenta of 13.5-day fetus was significantly increased in alcohol group,which was consistent with that in blastocyst.3)the sulfite sequencing(BSP)of genomic DNA in blastocyst and 13.5-day fetus showed that the level of genomic DNA methylation in the promoter region of Plagl1 decreased.The results of Plakoglobi n knockdown and overexpression in preimplantation embryos and cells suggest that the expression of Plagl1 is regulated by Plakoglobin expression and methylation in its promoter region as well as histone acetylation.To sum up,this study preliminarily elucidates the possible ways of the effect of alcohol in early embryos on placental development.That is,alcohol exposure will affect the expression of plakoglobin and epigenetic modification of imprinted gene Plagl1,change its expression,affect the structure and function of placenta,and then affect the growth and development of fetus.Although there is no data on conditional knockout of this Plagl1 in placenta in this study,because of its core role in the imprinted gene regulatory network,it can be inferred that Plagl1 plays an important role in the apparent effect of alcohol on placental development.This study will help to understand the developmental damage of embryo and placenta caused by alcohol,and provide ideas for further prevention and treatment of related diseases. |
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