Selection and Identification of Acute Proteins with Proteomic Techniques in Rhinencephalon of Cyprinus Carpio After Wounded Acutely

Abstract

A combined available approach of low osmosis and freeze-soluble with liquid nitrogen was used to disrupt the rhinencephalon tissue of Cyprinus carpio (CC). A centrifugal method with low speed was used to extract proteins in rhinencephalon tissue of CC (CCRT), and high-resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) map with 1200 protein spots was obtained. In this study, several techniques were used for extracting and separating proteome in CCRT. Three lysis solutions were prepared to disrupt the tissue in the fish, and used to extract proteome, water-soluble proteins and fat-soluble proteins, respectively. Both water-soluble proteins and fat-soluble protein from the CCRT were effectively separated by an optimized 2D-PAGE method with high resolution. With treating by 10% acetic acid, six acute proteins in CCRT were selected by proteomic techniques, and further identified by peptide mass fingerprinting (PMF) and database research, respectively. Three of these differential proteins can be primarily defined to be 70 kD heat-shock protein, DNA ligase IV, and β 5 tubulin. It was proposed that these proteins may be used as markers to study the repairing pathway and mechanism in brain once wounded.