Detection of Structural and Metabolic Changes in Traumatically Injured Hippocampus by Quantitative Differential Proteomics

• DOI: 10.1089/neu.2012.2391
• Publication Date: 2012-07-04T22:31:00Z
• Authors: Ping Wu; Yingxin Zhao; Sigmund J. Haidacher; Enyin Wang; Margaret O. Parsley; Junling Gao; Rovshan G. Sadygov; Jonathan M. Starkey; Bruce A. Luxon; Heidi Spratt; Douglas S. DeWitt; Donald S. Prough; Larry Denner
• Keywords: Male; Proteomics; Calcineurin; Blotting, Western; Hippocampus; Rats; 3. Good health; Rats, Sprague-Dawley; Disease Models, Animal; 03 medical and health sciences; 0302 clinical medicine; Tandem Mass Spectrometry; Brain Injuries; Animals; Chromatography, Liquid

Traumatic brain injury (TBI) is a complex and common problem resulting in the loss of cognitive function. In order to build comprehensive knowledge base proteins that underlie these deficits, we employed unbiased quantitative mass spectrometry, proteomics, bioinformatics identify quantify dysregulated CA3 subregion hippocampus fluid percussion model TBI rats. Using stable isotope 18O-water differential labeling multidimensional tandem liquid chromatography (LC)-MS/MS with high stringency statistical analyses filtering, identified quantified 1002 proteins, 124 increased 76 decreased. The ingenuity pathway analysis (IPA) tool had profound effects on downregulating global energy metabolism, including glycolysis, Krebs cycle, oxidative phosphorylation, as well cellular structure Widespread upregulation actin-related cytoskeletal dynamics was also found. IPA indicated integrative signaling node, calcineurin B1 (CANB1, CaNBα, or PPP3R1), which downregulated by TBI. Western blotting confirmed regulatory subunit, CANB1, its catalytic binding partner PP2BA, were decreased without changes other subunits. CANB1 plays critical role networks calcium homeostasis through calmodulin calmodulin-dependent kinase II highly interconnected structural dominated tubulins. This large-scale lays foundation for identification novel therapeutic targets rescue