斑马鱼个体发育过程中的溶菌酶表达模式及其对LPS处理的应答

Lysozyme expression pattern and its response to LPS treatment during zebrafish development process

收集斑马鱼成鱼和不同发育阶段幼鱼,用LPS处理后提取总RNA,经反转录制备cDNA并检测溶菌酶的基因表达水平,利用实时定量PCR技术研究了斑马鱼个体发育过程中溶菌酶的表达模式及其对LPS处理的应答方式.结果表明:成鱼体内溶菌酶基因表达水平显著高于各发育阶段幼鱼,而且LPS处理成鱼后溶菌酶表达水平呈现出先降低后升高的应答模式.在对照组幼鱼的不同发育阶段,溶菌酶基因表达水平变化不大.在个体发育早期(3d和7d),LPS处理可引起溶菌酶表达水平持续降低.14d和21d幼鱼中溶菌酶表达水平都在LPS处理后3h开始降低,并在6h开始回升,其中21d幼鱼回升到显著高于对照组的水平,表明溶菌酶已表现出对外界刺激作出有效应答的能力.另外,28d幼鱼受LPS处理后溶菌酶基因表达的变化趋势与成鱼相似,均为先降低后升高.以上结果说明,溶菌酶在斑马鱼个体发育到21d时可对外界刺激作出有效应答,在28d时则可能发育成熟.

Adult fish and j uvenile fish of zebrafish at different developmental stages were collected and treated with LPS,then,their total RNA were extracted and inverse transcribed to prepare cDNA,the gene expression level of lysozyme was also detected,and the lysozyme expression pattern and its response to LPS treatment during zebrafish development process were studied by real-time quantitative PCR.The results showed that the lysozyme expression level in adult fish was higher than that in j uvenile fish.After the adult fish was treated with LPS,the lysozyme expression level decreased firstly and then increased.No significant difference of the lysozyme expression was observed in the j uvenile fish at different development stages.LPS treatment reduced the lysozyme expression in the early development stages of zebrafish （3 d and 7 d）.The lysozyme expression level of 14-day-old and 21-day-old fish decreased at 3 h after treated with LPS and bot-tom out after 6 h,the difference was that lysozyme expression level in 2 1-day-old fish was much higher than that of control,this indicated that lysozyme showed the ability of effectively responding to the external stimulus.Moreover,in the juvenile fish of 28 d old treated with LPS,the variation trend of lysozyme was similar to that of the adult fish,both decreased firstly and then increased,the results indicated that the ly-sozyme might maturate since 2 8 d.