The Study Of Ascorbic Acid Improves Parthenogenetic Embryo Development Through TET Proteins In Mice

The TET?Ten-Eleven Translocation?proteins was originally named after a chromosomal translocation identified in patients with acute myeloid leukemia in the early 21st century and are?-ketoglutarate and Fe²⁺dependent dioxygenases??-KGDDs?that catalyze the oxidation of 5mC to 5hmC during early mammalian embryogenesis,which it's an important enzyme in the process of DNA demethylation and play crucial roles in embryonic development.After the sperm and oocyte of the mouse are combined,a series of epigenetic reprogramming will occur,in which a very important change in DNA methylation will occur,and the change in methylation status mainly occurs in the paternal genome,and the occurrence of the nail A process of basicization.5mC is a representative cytosine of whole genome methylation.If the gene is demethylated,the genome 5mC will oxidize to 5hmC.The expression levels of 5mC and 5hmC were used as the evaluation criteria for overall methylation.The above process of 5mC oxidation to 5hmC is mainly because the TET protein mainly converts 5mC into 5hmC by hydrogen bonding and stacking action of 5mC pyrimidine ring.Ascorbic acid?Vc,vitamin C?is a cofactor for Fe2+and?-KG-dependent dioxygenase and stimulates the expression of TET proteins,whereas DMOG?dimethylallyl glycine?is a structural analog of a 2-oxoglutarate cofactor that acts as a small molecule inhibitor of 2-oxoglutarate?2-OG?-dependent dioxygenase and inhibits TET expression.Epigenetic modification has a strong correlation with the early embryonic development potential of mice.DNA methylation is considered to be one of the epigenetic modifications involved in imprinting and is considered to be a major epigenetic modification.Abnormalities in methylation blots may be part of the reason for the failure of parthenogenetic embryo development.To investigate the role of TET1,TET2 and TET3 in parthenogenetic?PA?embryonic development,Vc and DMOG treatments were administered during early embryonic development.The results showed that Vc treatment increased the blastocyst rate?20.73±0.46%vs 26.57±0.53%?.It can significantly improve the developmental capacity of parthenogenetic embryos.By contrast,DMOG reduced the blastocyst rate?20.73±0.46%vs 11.18±0.13%?in PA embryos.Quantitative real-time PCR?qRT-PCR?and immunofluorescence?IF?staining results revealed that TET1,TET2,and TET3 expression were significantly lower in PA embryos compared with normal fertilized?Con?embryos.Our results revealed that Vc stimulated the expression of TET proteins in PA embryos.However,treatment with DMOG significantly inhibited the expression of TET proteins.It inhibits the development of parthenogeneticembryos.Weusedreal-timequantitativePCRand immunofluorescence staining to analyze the expression of 5mC and 5hmC in parthenogenetic embryos and the expression of three TET proteins.In addition,5hmC was increased following treatment with Vc and suppressed by DMOG in PA embryos.Taken together,these results indicate that the expression of TET proteins plays crucial roles mediated by 5hmC in PA embryonic development.