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目的通过大规模化学遗传学方法筛选斑马鱼髓系造血缺陷突变体。方法利用化学诱变剂乙基亚硝基脲(ENU)将雄鱼精原母细胞诱变,通过与AB野生型雌性斑马鱼交配产生F1代,源自不同founder(F0)的F1代同胞之间交配产生F2家族,F3代来F2家族同胞内交配,并分别以中性红和苏丹黑B染色筛选巨噬细胞和粒细胞缺陷突变体。结果我们筛选了350个F2家族,共1424对鱼。初步筛选到6个斑马鱼髓系造血系统的突变体,其中3个突变体为中性红染色异常,另3个突变体为苏丹黑染色异常,表明以上突变体可能存在巨噬细胞或粒细胞的发育障碍。结论 ENU化学诱变和中性红及苏丹黑B染色筛选斑马鱼髓系造血缺陷突变体是简单、价廉、有效的化学遗传学方法。通过保留突变体对后代进行进一步的研究分析,有望鉴定和克隆影响斑马鱼髓系造血新的基因或已知基因的新的调控途径。
Objective To screen zebrafish myeloid hematopoietic deficient mutants by large-scale chemical genetics. Methods The male spermatogonia were mutagenized with ethyl nitrosourea (ENU) as a chemical mutagen and F1 plants were generated by mating with AB wild-type female zebrafish. F1 progeny derived from different founder (F0) The F2 family was crossed and F3 families were crossed with the F2 family siblings. Macrophages and granulocyte-deficient mutants were screened by neutral red and Sudan black B respectively. Results We screened 350 F2 families for a total of 1424 pairs of fish. Preliminary screening of six zebrafish myeloid hematopoietic system of mutants, of which three were abnormal neutral red staining, the other three were Sudan black staining abnormalities, suggesting that the above mutants may exist macrophages or granulocytes Developmental disorders. Conclusion ENU chemical mutagenesis and neutral red and Sudan black B screening of zebrafish myeloid hematopoietic-deficient mutants are simple, inexpensive and effective chemical genetic methods. Further studies on the offspring by conserving mutants are expected to identify and clone new regulatory pathways that affect new genes or known genes in zebrafish myeloid hematopoiesis.