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Characterization of a weak allele of zebrafish cloche mutant
Authors:Ma Ning  Huang Zhibin  Chen Xiaohui  He Fei  Wang Kun  Liu Wei  Zhao Linfeng  Xu Xiangmin  Liao Wangjun  Ruan Hua  Luo Shenqiu  Zhang Wenqing
Institution:Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases of Guangdong Higher Education Institutes, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Abstract:Hematopoiesis is a complicated and dynamic process about which the molecular mechanisms remain poorly understood. Danio rerio (zebrafish) is an excellent vertebrate system for studying hematopoiesis and developmental mechanisms. In the previous study, we isolated and identified a cloche(172) (clo(172)) mutant, a novel allele compared to the original cloche (clo) mutant, through using complementation test and initial mapping. Here, according to whole mount in-situ hybridization, we report that the endothelial cells in clo(172) mutant embryos, although initially developed, failed to form the functional vascular system eventually. In addition, further characterization indicates that the clo(172) mutant exhibited weaker defects instead of completely lost in primitive erythroid cells and definitive hematopoietic cells compared with the clo(s5) mutant. In contrast, primitive myeloid cells were totally lost in clo(172) mutant. Furthermore, these reappeared definitive myeloid cells were demonstrated to initiate from the remaining hematopoietic stem cells (HSCs) in clo(172) mutant, confirmed by the dramatic decrease of lyc in clo(172)runx1(w84x) double mutant. Collectively, the clo(172) mutant is a weak allele compared to the clo(s5) mutant, therefore providing a model for studying the early development of hematopoietic and vascular system, as well as an opportunity to further understand the function of the cloche gene.
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