摘要
目的斑马鱼原始造血髓系细胞突变体1276的表型鉴定及性状分析。方法利用化学诱变剂N-乙基-N-亚硝基脲诱变野生型AB斑马鱼雄鱼的精原细胞,突变基因组保留传代,采用中性红染液对F3代的胚胎进行细胞化学染色,筛选原始造血髓系细胞突变体。并通过细胞化学染色及检测不同谱系血细胞标记基因在不同发育阶段的mRNA及蛋白的表达水平,对其中之一突变体1276进行表型鉴定及性状分析。结果成功筛选1296个F2家族,2140个突变基因组,发现中性红染色信号异常突变体12个。突变体1276在原始造血阶段,中性红染色全身信号缺失,小胶质细胞标记基因apoe表达缺失,巨噬细胞标记基因l-plastin,头部表达减少,背主动脉壁腹侧区域表达正常,粒细胞、红细胞均未见异常;在定向造血阶段,巨噬细胞仍存在异常,但粒细胞、红细胞、淋巴细胞以及造血干细胞均未见明显异常。结论突变体1276在原始造血阶段,巨噬细胞存在功能障碍,以及不同程度发育及迁徙异常;在定向造血阶段这一性状并未得到代偿。
Objective To perform phenotypic identification and characteristic analysis of a new zebrafish mutant 1276 defective in primitive myelopoiesis.Methods The AB strain male zebrafish were mutagenized with N-ethyl N-nitrosourea（ENU） to induce mutations in the spermatogonial cells,and the mutations were transmitted to the offsprings.The F3 embryos were screened by neutral red staining for identifying the mutants defective in primitive myelopoiesis.One of the myeloid mutants 1276 was further studied by cytochemistry and whole mount in stiu hybridization（WISH） with different lineage markers.Results A total of 2140 mutagenized genomes from the 1296 F2 families were analyzed,and 12 mutants were identified to show abnormal signal by neutral red staining.In the primitive hematopoiesis stage,the mutant 1276 showed the absence of neutral red staining-positive cells in the whole body.The expression of microglia marker apoe was totally lost in the head of the mutant,and the expression of the macrophage marker l-plastin was slightly decreased in the head and remained normal in the ventral dorsal aorta region,but the granulocytes and erythrocytes developed normally.in the definitive hematopoiesis stage,the mutant 1276 still showed abnormal macrophages as found in the primitive hematopoiesis stage,but the granulocytes,erythrocytes and lymphocytes appeared normal.Conclusion The zebrafish mutant 1276 shows abnormalities in the function,development and migration of the macrophages in the primitive hematopoiesis stage,which can not be compensated in the definitive hematopoiesis stage.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2011年第5期755-760,共6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)~~
