Forward genetic screening for zebrafish mutants defective in myelopoiesis / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To identify zebrafish mutants with myelopoiesis defects by ENU mutagenesis and large-scale forward genetic screening.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>