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TitleBioorthogonal 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.
KeywordsBioorthogonal chemistry
Neuronal development
Biorthogonal chemistry
Issue dateFeb-2020
CitationSoares 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.
DescriptionThe 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.
Publisher version
AccessOpen access
Appears in Collections:3B’s - Artigos em revistas/Papers in scientific journals
ICVS - Artigos em revistas internacionais / Papers in international journals

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