Screening the Differential Expressed Proteins by Protein Chip in Gill of Red Tilapia (Oreochromis mossambicus ♀ × O. niloticus ♂) Under Alkalinity Stress
Abstract Euryhalinous teleosts can survive in certain range saline-alkali water environment. The Gill is considered to be the main organ which is responsible for ion transport , maintain osmotic pressure balance and acid-base equilibrium.This study was to screen and identify the differential expressed proteins in gill of the Red tilapia (Oreochromis mossambicus ♀× O. niloticus ♂) by protein chips and mass spectrometry techniques, then verified by using immunohistochemistry and Western blot techniques, and to provide protein theoretical basis for alkali-resistant mechanism. There were 229 differential expressed protein spots detected (change multiple≥1.5), which contains 41 up-regulated proteins and 49 down-regulated proteins. Among them, 2 proteins were identified by mass spectrometry from 5 differential expressed proteins: 14-3-3 protein beta/alpha-A-like isoform X1 (14-3-3), Dna J homolog subfamily β member 11-like (Dna J).The immunohistochemistry results showed that 14-3-3 and Dna J were expressed in the gill base of Red tilapia at fresh water and alkalinity water, and showed a trend of first decrease and then the positive reaction increased significantly with increasing alkalinity. The results of Western blot showed that in the fresh water and alkali water, 14-3-3 and Dna J proteins were both expressed in the gill. With increasing alkalinity, the protein expression was the same as the immunohistochemistry. The expression of 14-3-3 protein showed a trend of increase with the increase of alkalinity concentration, and the expression of Dna J protein was significantly increased at alkalinity water which compare with fresh water. In the 4 alkalinity group , 14-3-3 protein reduced in the early stage of stress,then increased, and reached the lowest value at 12 h, and the peak value rised at 72 h. While the expression of Dna J protein was significantly increased at 96 h (P＜0.05). The results suggest that red tilapia is able to cope with protein activitity and stability in the body by activating the protein 14-3-3 and Dna J protein in the gill tissue, thereby maintaining the body protein normal function stability.The expression of 14-3-3 and Dna J proteins in gill of red tilapia was increased under alkalinity stress. Thus it would have great significance to support that 14-3-3 and Dna J proteins are both participated in the response of alkalinity stress.The present study gives more information to learn the mechanism of alkalinity stress response in red tilapia.
HUANG Sai-Ying,DIAO Jin-Liang,YU Yan, et al. Screening the Differential Expressed Proteins by Protein Chip in Gill of Red Tilapia (Oreochromis mossambicus ♀ × O. niloticus ♂) Under Alkalinity Stress[J]. , 2018, 26(12): 2100-2108.
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