ripply1在斑马鱼早期胚胎背腹发育中的作用

目的 探究ripply1基因在斑马鱼早期背腹轴发生过程中的作用。方法 利用斑马鱼整封原位杂交技术揭示ripply1基因在斑马鱼早期胚胎发育过程中的表达模式，通过显微注射技术在胚胎1细胞期注射ripply1的mRNA来高表达Ripply1蛋白并在后期观察胚胎背腹标记基因的变化及胚胎形态变化。利用Tol2转基因技术构建ripply1启动子驱动的GFP转基因鱼。结果 原位杂交结果显示ripply1基因在斑马鱼原肠早期胚盾期特异表达在胚盾处，即预定的背部。高表达ripply1后，在胚盾期背部标记基因表达范围扩大，腹部标记基因表达减弱，受精后24小时胚胎表现出严重的背部化表型：头部增大，腹部卵黄延伸减弱，尾部躯干及尾部区域减少，有的甚至形成了第二个体轴。得到的转基因鱼揭示ripply1母源表达，并且转录起始位点上游1200个碱基驱动的GFP能模拟内源基因的表达图式。结论 ripply1可能参与斑马鱼胚胎早期背腹轴的发生。

The role of ripply1 in zebrafish dorsal-ventral development

Objective To explore the role of ripply1 in zebrafish dorsal-ventral development. Methods Using zebrafish whole mount in situ hybridization to examine ripply1 expression pattern in early embryo development. Analyse the expression pattern change of dorsal-ventral marker genes at shield stage and the morphological change at 24hpf (hours post-fertilization) after overexpression of Ripply1 by injecting synthetic mRNA at 1 cell stage. Use of Tol2 transposon technology to obtain a ripply1 promoter driven GFP transgenic fish and to identify promoter region that recapitulates endogenous ripply1 expression pattern. Results In situ hybridization results reveal that ripply1 specifically expresses in the future dorsal region at shield stage. Overexpression of ripply1 causes an enlarged expression of dorsal marker genes and a reduction of ventral marker genes. Embryos overexpressing ripply1 also show severely dorsalized phenotype, with enlarged head, reduced ventral yolk extension, and shortened posterior trunk and tail regions, and the formation of a secondary trunk axis. Transgenic fish reveals the maternal expression of ripply1 and suggests that a 1.2kb promoter-driven GFP is able to recapitulate the endogenous gene expression pattern. Conclusion ripply1 may participate in zebrafish dorsal-ventral development.