Analysis Of Glycoproteins Involved In The Proliferation And Differentiation Of C17.2 Neural Stem Cells

Neural stem cells (NSCs), distributed in the nervous system, have the capability of self-renewal and multipotential, and under specific conditions, they can differentiate into neurons, astrocytes and oligodendrocytes. Since we know NSCs’ ability of self-proliferation and multipotential, we hope to see their therapeutic effect in such difficult cases as the damage of central nervous system and degenerative diseases. Due to their significant prospect of application, it has great clinical significance and scientific to study NSCs’ new surface markers as well as those proteins related to the self-renewal, proliferation and differentiation of NSCs. Glycosylation, as one of the most important post-translational modifications, has played an important role on the structure and functions of proteins. Research finds that the proliferation and differentiation of NSCs are regulated precisely by signaling pathway, in which process glycoproteins perform important physiological functions; therefore, large-scale, high-throughput study on the difference of glycoprotein expression and the variation of glycosylation by using quantitative glycoproteomic technology, which will contribute to the discovery of new molecules of NSCs’cell surface markers and key proteins involved in the proliferation and differentiation of NSCs, and it will also help to extend our understanding of the underline mechanisms of NSCs’ proliferation and differentiation. The objective of this experiment is to study the changes of glycoproteins in the process of astroglial differentiation of C17.2 neural stem cells. C17.2 neural stem cells, with the qualities of immortality, multipotential, and could be in vivo tracked, are good carriers for the in vitro study of neural stem cells at present. We made a quantitative analysis of glycoproteins of C17.2 neural stem cells differentiating into astrocytes after AICAR stimulation by means of SILAC (Stable isotope labeling by amino acids in cell culture) technology combined with lectin affinity and hydrazine chemistry. By mass spectrometry, we identify N-linked 123 glycosylated proteins, of which 57 glycoproteins have relative quantitative information and 16 cell plasma membrane glycoproteins display significant differences in the process of differentiating from NSCs to astrocytes. These glycoproteins play an important physiological role in cell adhesion, cell proliferation, cell differentiation and the process of nervous system development. Our research not only enlarges the dataset of glycoproteins, but also provides an important theoretical basis for discovering new neural cell surface markers, studying the proliferation and directed differentiation of NSCs, as well as the treatment of nervous system diseases.