选择一些可能参与斑马鱼性别决定相关的基因(cyp19a、cyp19b、gata4、dmrt1和wt1),通过RT—PCR和实时荧光定量PCR技术分析各个基因在斑马鱼64细胞期(2h)、256~512细胞期(2.5h)、囊胚球体期(4h)、30%外包(4.6h)、50%外包(5.2h)、75%外包(8h)、胚孔封闭期(10h)、10~15体节期(15h)、成型期10~20(32h)、孵化期(60h)、幼苗期(120h)以及7、10、12、14、16、18、20、22、24、26、28和30d的表达情况。结果显示:cyp19a与cyp19b基因随着胚胎的发育表达量逐渐增加,在28~30d达到最高;gata4基因在胚孔封闭期表达量最高,随后降低;dmrt1基因在30%外包期表达量最高,随后逐渐递减;wt1基因在10~15体节期表达量最高,随后逐渐递减并保持稳定的表达。
The expression level of genes (cyp19a, cyp19b, gata4, dmrt1 and wt1 ) which related to sex determination was profiled during different stages of embryogenesis of zebrafish(64 -cell, 256 -512 -cell, sphere, 30% - epiboly, 50% - epiboly, 75 % - epiboly, bud, 10 - 15 - somites, prim - 10 - 20, pec - fin, early larva, and 7, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 dpf (day-post-fertilization)) using reverse-transcription polymerase chain reaction (RT- PCR) and real-time PCR. The results indicated that the expression level of cyplga gene and cyp19b gene at 28dpf and 30dpf stage were significantly higher than other stages. The expression level of gata 4 gene was higher in gastrula-bud stage than other stages. The dmrt 1 gene was most expressed in 30% - epibolystage. The wtl gene was most expressed in 10 - 15 somites stage.