Bioorthogonal labeling reveals different expression of glycans in mouse hippocampal neuron cultures during their development

doi:10.3390/molecules25040795

Please use this identifier to cite or link to this item: https://hdl.handle.net/1822/64096

Title:	Bioorthogonal labeling reveals different expression of glycans in mouse hippocampal neuron cultures during their development
Author(s):	Costa, Diana Pereira Soares Sousa, João Carlos Mesquita, Sandro Gabriel Ferreira Dá Petkova, Nevena Ivanova Marques, Fernanda Reis, R. L. Sousa, Nuno Pashkuleva, I.
Keywords:	Bioorthogonal chemistry glycosylation Imaging Neuronal development Biorthogonal chemistry
Issue date:	Feb-2020
Publisher:	MDPI
Journal:	Molecules
Citation:	Soares da Costa D., Sousa J. C., da Mesquita S., Petkova-Yankova N., Marques F., Reis R. L., Sousa N., Pashkuleva I. Bioorthogonal Labeling Reveals Different Expression of Glycans in Mouse Hippocampal Neuron Cultures during Their Development, Molecules, Vol. 25, Issue 4, pp. 795, doi:10.3390/molecules25040795, 2020
Abstract(s):	The expression of different glycans at the cell surface dictates cell interactions with their environment and other cells, being crucial for the cell fate. The development of the central nervous system is associated with tremendous changes in the cell glycome that is tightly regulated. Herein, we have employed biorthogonal Cu-free click chemistry to image temporal distribution of different glycans in live mouse hippocampal neurons during their maturation in vitro. We show development-dependent glycan patterns with increased fucose and decreased mannose expression at the end of the maturation process. We also demonstrate that this approach is biocompatible and does not affect glycan transport although it relies on an administration of modified glycans. The applicability of this strategy to tissue sections unlocks new opportunities to study the glycan dynamics under more complex physiological conditions.
Type:	Article
Description:	The following are available online. Scheme S1: Synthesis of peracetylated azidomannose (Ac4ManNAz); Scheme S2: Alternative synthesis of GlcNAz using chloroacetic anhydride and NaOH as a base; Scheme S3: Synthesis of peracetylated azidofucose (Ac4FucAz); Figure S1: 1H-NMR spectra of ManNAz (D2O, 300 MHz); Figure S2: 1H-NMR spectra of Ac4ManNAz (CDCl3 , 300 MHz), mixture of anomers; Figure S3: HPLC chromatogram of purified Ac4ManNAz showing the two anomers; Figure S4: 1H-NMR spectra of GlcNAz (D2O, 300MHz); Figure S5: 1H-NMR spectra of Ac4GlcNAz (CDCl3 , 300 MHz), mixture of anomers; Figure S6: HPLC chromatogram of purified Ac4GlcNAz showing the two anomers; Figure S7: 1H-NMR spectra of 6-azido-1,2,3,4-tetra-O-acetyl-6-deoxy-α,β-L-galactopyranose Ac4FucAz (CDCl3 , 300 MHz): mixture of anomers; Table S1: Primer sequences used in qRT-PCR.
URI:	https://hdl.handle.net/1822/64096
DOI:	10.3390/molecules25040795
ISSN:	1420-3049
Publisher version:	https://www.mdpi.com/1420-3049/25/4/795
Peer-Reviewed:	yes
Access:	Open access
Appears in Collections:	3B’s - Artigos em revistas/Papers in scientific journals ICVS - Artigos em revistas internacionais / Papers in international journals