Dissecting the Role of O-GlcNAc in Mouse Preimplantation Development

Chitnavis, Urvashi Milind (2022) Dissecting the Role of O-GlcNAc in Mouse Preimplantation Development. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Urvashi Milind Chitnavis (17MS051)) 17MS051_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (1MB)

Abstract

O-linked β-N-Acetylglucosamine (O-GlcNAc) is a protein-carbohydrate linkage found on serine and threonine residues of a variety of cytosolic and nuclear proteins. O-GlcNAc and the protein that catalyses this modification, O-GlcNAc transferase (OGT), have been widely studied in a range of cell types and metabolic contexts. However, despite evidence of a critical role for OGT in mammalian embryonic development, the context and mechanism of its function during this crucial time window remains unknown. In this study, we attempt to elucidate the role of OGT and O-GlcNAc during the preimplantation period of mammalian embryonic development - beginning in the zygote, through embryonic genome activation at the 2-cell stage, until the blastocyst. Due to the evolutionary proximity of mice to humans and the ability to extend information obtained in the mouse to understanding general mammalian development, we study OGT and O-GlcNAc in the developing mouse embryo as well as in mouse embryonic stem cell cultures. We use techniques ranging from embryo immunofluorescence and microinjections, to siRNA-based knockdown, western blotting and flow cytometry, to formulate an understanding of the function of O-GlcNAc in the preimplantation embryo. We find that OGT and O-GlcNAc have distinct distribution and functions during the various developmental stages of the mouse embryo, and our results indicate several lines of experimentation that will further elucidate their role in mammalian embryonic gene regulation.

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