斑马鱼红系造血缺陷突变体的正向遗传学筛选  

Forward genetic screening for zebrafish mutants defective in erythropoiesis

作　　者：霍中军[1] 温宗华[2] 刘靖[1] 王鹍[1] 黄志斌[1] 戴朝霞[1] 马宁[1] 颜广[1] 陈英华[3] 陈小辉[1] 刘伟[1] 马品芸[4] 罗伟豪[4] 赵颖[4] 樊淑[4] 赵嘉佳[1] 黄红辉[4] 温子龙[5] 张文清[1] 

机构地区：[1]南方医科大学基础医学院细胞生物学教研室,广东广州510515 [2]南方医科大学基础医学院病理学教研室,广东广州510515 [3]南方医科大学基础医学院病组织胚胎学教研室,广东广州510515 [4]南方医科大学基础医学院医学遗传学教研室,广东广州510515 [5]香港科技大学生化系,香港

出　　处：《南方医科大学学报》2010年第5期931-935,共5页Journal of Southern Medical University

基　　金：国家自然科学基金海外及港澳学者合作研究项目(30828020)

摘　　要：目的化学遗传学方法大规模筛选并初步鉴定所获得具有不同红系造血缺陷表型的斑马鱼突变体。方法乙基亚硝基脲(ENU)诱导雄性斑马鱼突变(founder),将其与野生型AB雌性斑马鱼交配产生F1代,源自不同来源的founder的F1代杂交产生F2家族。在F2代同家族内自交所产生的F3代胚胎中,用以βe1为探针,实施整体原位杂交实验,进行红系造血缺陷突变体筛选,并针对所筛选到的突变体在不同造血过程缺陷表型进行分类研究。结果和结论筛选得到4个βe1基因表达缺失突变体,其中2个为红系特异性造血缺陷突变体,另外2个突变体同时存在红系和淋系造血缺陷。Objective To screen and identify zebrafish mutants with erythropoiesis defects by N-ethyl-N-nitrosourea （ENU） mutagenesis and large-scale forward genetic screening using βe1 as the marker. Methods The chemical mutagen ENU was used to treat healthy wild-type male fish （AB strain, F0）. The surviving ENU-treated fish were mated with wild-type female fish to generate F1, and further F2 family was generated by F1 family intercross. The adult F2 fish were intercrossed within each F2 family and the resulting F3 embryos from each crossing were subjected to whole mount in situ hybridization （WISH） with the βe1 probe. Mutagenesis was performed by treating the male zebrafish with ENU to induce mutations in pre-meiotic germ cells to generate the founders, which were outcrossed to obtained the F1 fish. The F1 fish from different founders were mated to generate the F2 families. F3 embryos from the sibling cross in the F2 family were examined by whole mount in situ hybridization using βe1-globin probe. The putative mutants were then characterized with different hematopoiesis markers. Results and Conclusion We identified 4 βe1-deficient mutants with erythropoiesis defects, including two with specific erythiod lineage defects and two with concurrent lymphopoiesis defects.

关 键 词：斑马鱼 红系造血 正向遗传学筛选 βe1-globin 

分 类 号：R-332[医药卫生]