Abstract：Establishing a Salmonella-zebrafish infection model to study the xenophagy. Zebrafish larvae at 72 h post-fertilization were exposed to different doses of Salmonella typhimurium (S. typhimurium). The survival curve of zebrafish larvae was used to determine the appropriate dose of bacteria. After infection by the above dose, the distribution of green fluorescent protein (GFP)-tagged S. typhimurium in zebrafish larvae was observed under a fluorescence microscope. The bacteria count and expression of autophagy protein Lc3-Ⅱand P62 were used to detect the changes in autophagy, and the autophagosome was observed under the transmission electron microscope. The results revealed that after exposure to S. typhimurium at a final dose of 1×109 cfu/ml, the survival curve was 100%, and the larvae began to die on the third day. Fluorescence microscopy found that bacteria had invaded into the larvae at 4 h after infection, and spreaded around the body in 3 d. The bacteria amount in whole-larvae bacteria was increasing over time, but the amount of intracellular bacteria in the larvae at 10 h after infection was significantly less than that at 8 h. Western blotting analysis showed that Lc3-Ⅱ was significantly increased, but P62 was reduced. Under the transmission electron microscope, autophagosome and autolysosome were observed in larvae at 8 h after infection. According to the above results, Salmonella-zebrafish model can be used to track the dynamic changes in autophagy induced by pathogen infection.
李金玲,王婷,李嫄渊,吴淑燕,黄瑞. 沙门菌-斑马鱼感染模型用于自噬的探究[J]. 微生物与感染, 2014, 9(2): 77-82.
LI Jin-Ling, WANG Ting, LI Yuan-Yuan, WU Shu-Yan, HUANG Rui. Salmonella-zebrafish infection model used in autophagy. JOURNAL OF MICROBES AND INFECTIONS, 2014, 9(2): 77-82.
