Neuro-Compatible Metabolic Glycan Labeling of Primary Hippocampal Neurons in Noncontact, Sandwich-Type Neuron Astrocyte Coculture

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Glycans are intimately involved in several facets of neuronal development and neuropathology. However, the metabolic labeling of surface glycans in primary neurons is a difficult task because of the neurotoxicity of unnatural monosaccharides that are used as a metabolic precursor, hindering the progress of metabolic engineering in neuron-related fields. Therefore, in this paper, we report a neurosupportive, neuron astrocyte coculture system that neutralizes the neurotoxic effects of unnatural monosaccharides, allowing for the long-term observation and characterization of glycans in primary neurons in vitro. Polysialic acids in neurons are selectively imaged; via the metabolic labeling of sialoglycans with peracetylated Nazidoacetyl-o-mannosamine (Ac(4)ManNAz), for up to 21 DIV. Two-color labeling shows that neuronal activities, such as neurite outgrowth and recycling of membrane components, are highly dynamic and change over time during development. In addition, the insertion sites of membrane components are suggested to not be random, but be predominantly localized in developing neurites. This work provides a new research platform and also suggests advanced 3D systems for metabolic -labeling studies of glycans in primary neurons.
Publisher
AMER CHEMICAL SOC
Issue Date
2017-12
Language
English
Article Type
Article
Keywords

CELL-ADHESION MOLECULE; CENTRAL-NERVOUS-SYSTEM; POLYSIALIC ACID; IN-VIVO; SIALIC ACIDS; SURFACE GLYCANS; STEM-CELLS; BIOSYNTHESIS; EXPRESSION; PLASTICITY

Citation

ACS CHEMICAL NEUROSCIENCE, v.8, no.12, pp.2607 - 2612

ISSN
1948-7193
DOI
10.1021/acschemneuro.7b00300
URI
http://hdl.handle.net/10203/239476
Appears in Collection
CH-Journal Papers(저널논문)
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