Isolation and identification of differentially expressed proteins in hippocampus of mice during hypobaric hypoxic delayed preconditioning / 中国应用生理学杂志

ABSTRACT

<p><b>AIM</b>To explore the differentially expressed proteins between hypobaric hypoxic delayed preconditioning (HHDP) and normal mouse hippocampus.</p><p><b>METHODS</b>After the animal model of HHDP was constructed, hippocampal proteins were obtained by a series of abstraction with lysis solution containing high concentration urea. As soon as isoelectric focusing and SDS-PAGE was performed. The resolved proteins in the 2-DE gels were visualized by Coomassie blue R-250. The gels were scanned, and the images were processed with PDQuest software. Differential proteins were exactly excised from the gels, destained and digested with trypsin. The peptides were isolated and sent for MALDI-TOF-MS testing. Database searching was performed using peptide masses obtained from MALDI-TOF-MS.</p><p><b>RESULTS</b>Averages of 481 +/- 38 and 477 +/- 21 protein spots were detected in control gels and preconditioning gels, respectively. 169 +/- 6 protein spots were matched between these two types of gels. Among the matched spots, while the quantities of 21 +/- 12 spots in control gels increased by above 2 times than that in preconditioning one, the quantities of 33 +/- 10 spots in preconditioning gels increased by the same times than that in control one. The correlation coefficient between these two patterns were 0.7748 +/- 0.0267. 12 spots in preconditioning gels significantly increased compared with the control (P < 0.05, n = 4). Among 12 spots excised from the gels, perfect peptide mass fingerprinting spectrums of 8 spots were acquired. The results showed that one protein was fructose biphosphate aldolase A. Three proteins matched nothing might be new proteins. The other four proteins just matched the partial sequences of the proteins of database were no coincidence to it's isoelectric point and molecular weight. So they might be homological proteins.</p><p><b>CONCLUSION</b>Many proteins, for example fructose biphosphate aldolase A, has been differentially expressed in hippocampus of mice during HHDP. This may be one of the molecule mechanisms of HHDP.</p>