Two-dimensional electrophoresis analysis for protein profile change in zebrafish alcohol syndrome model
10.3760/cma.j.issn.1007-9408.2011.05.006
- VernacularTitle:二维电泳分析胎儿酒精综合征斑马鱼模型蛋白表达谱
- Author:
Linxi QIAN
;
Shuna SUN
;
Wei CAI
;
Yuexiang WANG
;
Qiu JIANG
;
Houyan SONG
- Publication Type:Journal Article
- Keywords:
Zebrafish;
Fetal alcohol syndrome;
Proteome;
Electrophoresis,gel,two-dimensional
- From:
Chinese Journal of Perinatal Medicine
2011;14(5):283-288
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the putative mechanisms underlying fetal alcohol syndrome by comparative protein-profile analysis between normal and ethanol-treated zebrafish embryo with twodimensional electrophoresis (2-DE).Methods Zebrafish embryos were exposed in 400 mmol/L ethanol at dome stage for 3 hours,and then ethanol-induced abnormalities were observed.Proteomes of zebrafish embryos at early stages including zygote stage,dome stage,shield stage and 5-somite stage,were separated by 2-DE.The subtraction analysis method was applied to eliminate the interference from maternal derived proteins.The ethanol-treated embryos at 5-somite stage was analyzed by 2-DE,and the protein profile was compared with that generated from control embryos at the same stage.The data obtained from 2-DE analysis were verified by in-situ hybridization.Results 400 mmol/L ethanol treatment caused axial malformation (62%) and cyclopia (60%) in zebrafish embryos.The 2-DE analysis showed that the expression of Collagen2al (Col2a1) and TAR DNA binding protein (TDP) was decreased in 12 hours post fertilization (12 hpf) ethanol-treated embryos by 81% and 73%,respectively.The in-situ hybridization also demonstrated that the expression of Col2al in axial mesoderm was reduced by ethanol treatment at the same stage.But for 24 hpf ethanoltreated embryos,the expression of Col2al in axis recovered to a comparable level to that in control embryos,while the structure of neural tube was disrupted severely by ethanol exposure.Conclusions It is suggested that the expressions of Col2al and TDP were disrupted by ethanol during early stage,which might induce the zebrafish developmental abnormalities.The ethanol interference on early expression of Col2al is supposed to be one of the major reasons leading to later abnormalities of axis and neutral tube.
