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Exploring O‐GlcNAcylation and phosphorylation in the developing chick brain
Author(s) -
Farach Andrew M.,
Galileo Deni S.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a57-b
Subject(s) - neurofilament , intermediate filament , vimentin , biology , microbiology and biotechnology , axon , intermediate filament protein , phosphorylation , western blot , cytoskeleton , neuroscience , immunohistochemistry , cell , biochemistry , immunology , gene
Interest in modification of proteins by O‐linked beta‐N‐acetylglucosamine (O‐GlcNAc) is growing rapidly due to its continued implication in the regulation of cellular processes. Since roles for O‐GlcNAc in brain development have not yet been described, we characterized the expression of O‐GlcNAc in the developing chick midbrain (optic tectum) to see if patterns existed in specific cell types, in subcellular processes like axons, or in specific proteins. Based on the recurrent O‐GlcNAcylation of highly phosphorylated proteins in other vertebrate systems, neurofilament modification in neuronal axons was examined. Our studies indicate that a regulated developmental pattern of O‐GlcNAcylation exists in the developing chick brain. Immunohistochemical analyses in sections of progressive stages of development suggest upregulation of O‐GlcNAc in the ependyma, tectobulbar neuron bodies, and radial glial processes, but not in neurofilament (+) axons. In contrast, O‐GlcNAcylation of most axons occurred in cultures made from embryonic day 7 brain cells. Western blot analysis showed O‐GlcNAc modification of a few discrete proteins throughout development, including one of approximate weight of vimentin (52kDa), an intermediate filament marker of radial glia in the chick brain. These results and labeling of radial glia in brain sections indicate that it is vimentin that is modified by O‐GlcNAcylation. This potentially indicates an undescribed developmental role for O‐GlcNAc modification that could include vimentin intermediate filaments as well as neurofilaments. Funded by HHMI, Charles Peter White Fellowship, and UD Undergraduate Research Program.