Dai Zhao-xia, Yan Guang, Chen Ying-hua, Liu Wei, Huo Zhong-jun, Wen Zong-hua, Liu Jing, Wang Kun, Huang Zhi-bing, Ma Ning, Chen Xiao-hui, Ma Ping-yun, Luo Wei-hao, Zhao Ying, Fan Shu, Huang Hong-hui, Wen Zi-long, Zhang Wen-qing
Department of Cell Biology, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Jun;30(6):1230-3.
To identify zebrafish mutants with myelopoiesis defects by ENU mutagenesis and large-scale forward genetic screening.
Male zebrafish were mutagenized with N-ethyl N-nitrosourea to induce mutations in the spermatogonial cells to generate the founders, which were outcrossed with AB to raise F1 fish. The F1 fish from different founders were mated to generate the F2 families. The F3 embryos from F2 sibling crosses were screened by Sudan black B staining and neutral red staining.
A total of 350 F2 families from F1 sibling crosses were screened, and 1424 F2 crosses were analyzed. Six mutations were identified resulting in abnormal Sudan black B staining and neutral red staining, indicating the involvement of neutrophil deficiency or macrophage abnormalities.
It is simple and cheap to induce and screen myelopoiesis deficiency in zebrafish by ENU chemical mutagenesis and Sudan black B staining and neutral red staining. These mutants shed light on the identification of the genes important to myelopoiesis in zebrafish.
通过ENU诱变和大规模正向遗传筛选鉴定具有髓系造血缺陷的斑马鱼突变体。
用N-乙基-N-亚硝基脲诱变雄性斑马鱼，诱导精原细胞发生突变以产生奠基者，将其与AB品系杂交培育F1代鱼。来自不同奠基者的F1代鱼相互交配产生F2家系。对F2同胞杂交产生的F3胚胎进行苏丹黑B染色和中性红染色筛选。
共筛选了来自F1同胞杂交的350个F2家系，分析了1424次F2杂交。鉴定出6个突变，导致苏丹黑B染色和中性红染色异常，表明存在中性粒细胞缺乏或巨噬细胞异常。
通过ENU化学诱变以及苏丹黑B染色和中性红染色在斑马鱼中诱导和筛选髓系造血缺陷既简单又经济。这些突变体有助于鉴定斑马鱼中对髓系造血重要的基因。
