title: The use of proteomic analysis to study trafficking defects in axons.
abstract: Mutations in microtubule subunits and microtubule-associated proteins are the causes of many neurological disorders. These human conditions are usually associated with axonal tract defects or degeneration. The molecular mechanisms of these axonal dysfunction are still largely unknown. Conventional methods may not yield a complete analysis of downstream molecules related to axonal dysfunctions. Therefore, we devised a simple unbiased method to screen molecular motors and axonal molecules, which might be involved in axonal defects. We performed our analysis in the mouse with a targeted deletion in the doublecortin (Dcx) gene. Dcx is a microtubule-associated protein with direct effects on microtubule motors. Furthermore, the knockout of Dcx and its functionally redundant structurally similar paralog, doublecortin-like kinase 1 (Dclk1), in mouse results in thinner or absent axon tracts, including the corpus callosum and anterior commissures. We compared protein profiles of corpus callosum from Dcx knockout and wild-type mouse of P0-P2 using mass spectrometry. This strategy allowed us to identify novel candidates downstream of Dcx involved in axon transport.
authors.affiliation: Center for Neuroscience Research, Children's National Health System, Washington, DC, USA.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Fu
authors.affiliation: Research Center for Genetic Medicine, Children's National Health System, Washington, DC, USA; Department of Integrative Systems Biology, Institute of Biomedical Sciences, The George Washington University, Washington, DC, USA.
authors.forename: Kristy J
authors.initials: KJ
authors.lastname: Brown
authors.affiliation: Research Center for Genetic Medicine, Children's National Health System, Washington, DC, USA; Department of Integrative Systems Biology, Institute of Biomedical Sciences, The George Washington University, Washington, DC, USA.
authors.forename: Sree
authors.initials: S
authors.lastname: Rayavarapu
authors.affiliation: Research Center for Genetic Medicine, Children's National Health System, Washington, DC, USA; Department of Integrative Systems Biology, Institute of Biomedical Sciences, The George Washington University, Washington, DC, USA.
authors.forename: Kanneboyina
authors.initials: K
authors.lastname: Nagaraju
authors.affiliation: Center for Neuroscience Research, Children's National Health System, Washington, DC, USA.
authors.forename: Judy S
authors.initials: JS
authors.lastname: Liu
mesh_terms: D000818:Animals; D001370:Axonal Transport / Q000235:genetics*; D001369:Axons / Q000378:metabolism* / Q000473:pathology; D002478:Cells, Cultured; D003337:Corpus Callosum / Q000166:cytology / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D000089742:Doublecortin-Like Kinases; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D009479:Neuropeptides / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D021381:Protein Transport / Q000235:genetics*; D040901:Proteomics / Q000379:methods
chemical_list: C000714328:DCX protein, human; C000714847:Dcx protein, mouse; D000089622:Doublecortin Domain Proteins; D000089662:Doublecortin Protein; D008869:Microtubule-Associated Proteins; D009479:Neuropeptides; D000089742:Doublecortin-Like Kinases; C485525:Dclk1 protein, mouse; D017346:Protein Serine-Threonine Kinases
keywords: Corpus callosum; Doublecortin; Lissencephaly; Mass spectrometry; Proteomics