中国实验动物学报
中國實驗動物學報
중국실험동물학보
Acta Laboratorium Animalis Scientia Sinica
2015年
5期
446-452
,共7页
斑马鱼%ripply1%早期胚胎发育%背腹发生
斑馬魚%ripply1%早期胚胎髮育%揹腹髮生
반마어%ripply1%조기배태발육%배복발생
Zebrafish%ripply1%Early embryo development%Dorsal-ventral axis
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.
目的 探究ripply1基因在斑馬魚早期揹腹軸髮生過程中的作用. 方法 利用斑馬魚整封原位雜交技術揭示ripply1基因在斑馬魚早期胚胎髮育過程中的錶達模式,通過顯微註射技術在胚胎1細胞期註射ripply1的mRNA來高錶達Ripply1蛋白併在後期觀察胚胎揹腹標記基因的變化及胚胎形態變化. 利用Tol2轉基因技術構建ripply1啟動子驅動的GFP轉基因魚. 結果 原位雜交結果顯示ripply1基因在斑馬魚原腸早期胚盾期特異錶達在胚盾處,即預定的揹部. 高錶達ripply1後,在胚盾期揹部標記基因錶達範圍擴大,腹部標記基因錶達減弱,受精後24小時胚胎錶現齣嚴重的揹部化錶型:頭部增大,腹部卵黃延伸減弱,尾部軀榦及尾部區域減少,有的甚至形成瞭第二箇體軸. 得到的轉基因魚揭示ripply1母源錶達,併且轉錄起始位點上遊1200箇堿基驅動的GFP能模擬內源基因的錶達圖式. 結論 ripply1可能參與斑馬魚胚胎早期揹腹軸的髮生.
목적 탐구ripply1기인재반마어조기배복축발생과정중적작용. 방법 이용반마어정봉원위잡교기술게시ripply1기인재반마어조기배태발육과정중적표체모식,통과현미주사기술재배태1세포기주사ripply1적mRNA래고표체Ripply1단백병재후기관찰배태배복표기기인적변화급배태형태변화. 이용Tol2전기인기술구건ripply1계동자구동적GFP전기인어. 결과 원위잡교결과현시ripply1기인재반마어원장조기배순기특이표체재배순처,즉예정적배부. 고표체ripply1후,재배순기배부표기기인표체범위확대,복부표기기인표체감약,수정후24소시배태표현출엄중적배부화표형:두부증대,복부란황연신감약,미부구간급미부구역감소,유적심지형성료제이개체축. 득도적전기인어게시ripply1모원표체,병차전록기시위점상유1200개감기구동적GFP능모의내원기인적표체도식. 결론 ripply1가능삼여반마어배태조기배복축적발생.
Objective To explore the role of ripply1 in zebrafish dorsal-ventral development .Methods Using ze-brafish whole-mount in situ hybridization to examine the ripply1 expression pattern in early embryo development .To analyse the expression pattern changes of dorsal-ventral marker genes at shield stage and the morphological changes at 24 hpf (hours post-fertilization) after overexpression of ripply1 by injecting synthetic mRNA at 1-cell stage.Using Tol2 transposon technology to obtain a ripply1 promoter driven GFP transgenic fish and to identify promoter region that recapitulates endoge -nous ripply1 expression pattern .Results The in situ hybridization results revealed that ripply1 specifically expresses in the future dorsal region at shield stage .Overexpression of ripply1 caused an enhanced expression of dorsal marker genes and a reduction of ventral marker genes .Embryos overexpressing ripply1 also showed 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 revealed the maternal expression of ripply1 and suggested that a 1.2 kb promoter-driven GFP is able to recapitulate the endogenous gene expression pattern .Conclusion ripply1 may participate in the early development of dor-sal-ventral axis in zebrafish embryo .
