Tissue-based metabolic labeling of polysialic acids in living primary hippocampal neurons.
Proc Natl Acad Sci U S A
; 112(3): E241-8, 2015 Jan 20.
Article
in En
| MEDLINE
| ID: mdl-25564666
ABSTRACT
The posttranslational modification of neural cell-adhesion molecule (NCAM) with polysialic acid (PSA) and the spatiotemporal distribution of PSA-NCAM play an important role in the neuronal development. In this work, we developed a tissue-based strategy for metabolically incorporating an unnatural monosaccharide, peracetylated N-azidoacetyl-D-mannosamine, in the sialic acid biochemical pathway to present N-azidoacetyl sialic acid to PSA-NCAM. Although significant neurotoxicity was observed in the conventional metabolic labeling that used the dissociated neuron cells, neurotoxicity disappeared in this modified strategy, allowing for investigation of the temporal and spatial distributions of PSA in the primary hippocampal neurons. PSA-NCAM was synthesized and recycled continuously during neuronal development, and the two-color labeling showed that newly synthesized PSA-NCAMs were transported and inserted mainly to the growing neurites and not significantly to the cell body. This report suggests a reliable and cytocompatible method for in vitro analysis of glycans complementary to the conventional cell-based metabolic labeling for chemical glycobiology.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Sialic Acids
/
Hippocampus
/
Neurons
Limits:
Animals
Language:
En
Journal:
Proc Natl Acad Sci U S A
Year:
2015
Type:
Article