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Location and Partners of Polysialic Acid in the Dorsal Horn and Trigeminal Nucleus of Adult Rat
Author(s) -
Shahbazian Shila,
Lee Albert,
EverestDass Arun,
Bokiniec Phill,
Packer Nicolle,
Goodchild Ann
Publication year - 2016
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.30.1_supplement.1096.1
Subject(s) - polysialic acid , western blot , neural cell adhesion molecule , nucleus , microbiology and biotechnology , chemistry , immunohistochemistry , neuroscience , biology , cell , biochemistry , immunology , cell adhesion , gene
Polysialic acid (PSA) is a large cell‐surface glycan predominantly attached to the neural cell adhesion molecule (NCAM) in the mammalian brain. PSA has discrete expression patterns to specific areas of the CNS, postulated to have high levels of synaptic plasticity. The regulation, expression and interacting partners of PSA is complex and remains poorly understood. In this study, we aimed to examine the cellular distribution of PSA by immunohistochemistry in the dorsal horn and trigeminal nucleus of adult Sprague Dawley rats. Both regions receive significant sensory input thus are likely to have high levels of plasticity. We further investigated interacting partners of PSA in these regions using co‐immunoprecipitation (IP) followed by label‐free liquid chromatography tandem mass spectrometry (LC‐MS/MS) analysis. We found that PSA was highly expressed in the superficial laminae of the dorsal horn, and within the trigeminal complex distribution was restricted to only the superficial layers of nucleus caudalis (TNc). Using LC‐MS/MS analysis, 14 proteins were identified in the dorsal horn as potential binding partners of PSA‐carrying NCAM. Three candidates (Receptor expression‐enhancing protein 5 (REEP5), Guanine nucleotide‐binding protein G(o) subunit alpha (GNOA1), and Clathrin heavy chain 1 (CLTC) were validated by IP and western blot analyses. Moreover, co‐IP of PSA from TNc preparations followed by western blot analyses of the three candidates confirmed their PSA status within TNc. This is the first study to demonstrate the detailed cellular distribution of PSA in the rat trigeminal nucleus and the identities of three potential interacting partners. These findings will improve our understanding of the mechanisms by which PSA affects synaptic plasticity and may reveal novel PSA functions. Support or Funding Information Anti‐polysialic acid antibody (mAb735) was kindly provided by prof. Rita Gerardy‐Schahn.