Abstract
Glycosylation is one of the most fundamental post-translational modifications. However, the glycosylation patterns of glycoproteins have not been analyzed in mammalian preimplantation embryos, because of technical difficulties and scarcity of the required materials. Using high-throughput lectin microarrays of low-input cells and electrochemical techniques, an integration analysis of the DNA methylation and glycosylation landscapes of mammal oogenesis and preimplantation embryo development was performed. Highly noticeable changes occurred in the level of protein glycosylation during these events. Further analysis identified several stage-specific lectins including LEL, MNA-M, and MAL I. It was later confirmed that LEL was involved in mammalian oogenesis and preimplantation embryogenesis, and might be a marker of FGSC differentiation. Modified nanocomposite polyaniline/AuNPs were characterized by electron microscopy and modification on bare gold electrodes using layer-by-layer assembly technology. These nanoparticles were further subjected to accuracy measurements by analyzing the protein level of ten-eleven translocation protein (TET), which is an important enzyme in DNA demethylation that is regulated by O-glycosylation. Subsequent results showed that the variations in the glycosylation patterns of glycoproteins were opposite to those of the TET levels. Moreover, analysis of correlation between the changes in glyco-gene expression and female germline stem cell glycosylation profiles indicated that glycosylation was related to DNA methylation. Subsequent integration analysis showed that the trend in the variations of glycosylation patterns of glycoproteins was similar to that of DNA methylation and opposite to that of the TET protein levels during female germ cell and preimplantation embryo development. Our findings provide insight into the complex molecular mechanisms that regulate human embryo development, and a foundation for further elucidation of early embryonic development and informed reproductive medicine.
Highlights
Post-translational modification refers to the covalent processing of translated proteins
Because it is difficult to obtain a large number of embryos, we first established lectin microarrays for low-input cells using SIM mouse embryo-derived thioguanine and ouabain-resistant (STO) feeder cells
The results showed that the background signal of the microarray sealed with protein-free sealing solution was low, and the intensity and signal-to-noise ratio of the positive signal in the sample were high (Supplementary Figures S2A–C)
Summary
Post-translational modification refers to the covalent processing of translated proteins. Glycosylation is one of the most important post-translational modifications It is the process by which carbohydrate residues, or glycans, are attached to biomolecules to produce glycoconjugates, and is the primary form of protein and lipid post-translational modifications (Spiro, 2002). Glycoproteins are widely distributed in organisms and involved in the regulation of many biological processes, including mammal oogenesis and preimplantation embryo development (PED). The dynamic patterns of epigenetic regulation have been analyzed, including DNA methylation (Guo et al, 2014), histone modification (Liu et al, 2016), non-coding RNAs (Fan et al, 2015), and 3D chromatin structures (Ke et al, 2017), the glycosylation patterns of glycoproteins have not been investigated in mammalian oogenesis and preimplantation embryo development because of technical difficulties and the scarcity of required materials
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