论文部分内容阅读
目的观察外源性视黄酸对斑马鱼心脏发育的影响,探讨视黄酸对 Tbx1基因在斑马鱼胚胎发育中表达的影响。方法采用化学遗传学的方法在斑马鱼胚胎发育至胚胎受精后12.5 h时给予5×10~(-8)mol/L 和10~(-7)mol/L 外源性视黄酸处理1.5 h。采用胚胎整体原位杂交和实时荧光定量 PCR 方法观察外源性视黄酸对斑马鱼胚胎发育过程中 Tbx1基因时空表达模式的影响。结果整体原位杂交显示 Tbx1在斑马鱼胚胎发育过程中主要在咽弓、心脏和耳囊表达。在12.5 hpf 给予斑马鱼胚胎5×10~(-8)mol/L 或10~(-7)mol/L 视黄酸处理1.5 h,可以导致 Tbx1基因在咽弓和心脏的表达明显下调。与5×10~(-8)mol/L 视黄酸处理组相比,10~(-7)mol/L 视黄酸对 Tbx1基因表达的抑制作用更为明显。结论视黄酸可以调节斑马鱼发育过程 Tbx1基因的表达,且这种调控作用具有明显的剂量依赖性。视黄酸是导致胚胎发育过程中 Tbx1表达异常的主要非遗传因素,并可进一步导致咽弓和心脏的发育异常。
Objective To observe the effect of exogenous retinoic acid on zebrafish heart development and to explore the effect of retinoic acid on the expression of Tbx1 in zebrafish embryos. Methods The chemical genetics method was used to treat exogenous retinoic acid at a dose of 5 × 10 ~ (-8) mol / L and 10 ~ (-7) mol / L for 1.5 h after the development of zebrafish embryo to 12.5 h after embryo fertilization . The effects of exogenous retinoic acid on the spatial and temporal expression pattern of Tbx1 gene during zebrafish embryogenesis were observed by whole-mount in situ hybridization and real-time fluorescence quantitative PCR. Results The overall in situ hybridization showed that Tbx1 was mainly expressed in pharyngeal arch, heart and ear during zebrafish embryogenesis. Treatment with zebrafish embryos at 5 × 10 -8 mol / L or 10 -7 mol / L retinoic acid for 12.5 h at 12.5 hpf resulted in a significant down-regulation of Tbx1 gene expression in pharyngeal arch and heart. Compared with 5 × 10 ~ (-8) mol / L retinoic acid treatment group, the inhibitory effect of 10 -7 mol / L retinoic acid on Tbx1 gene expression was more obvious. Conclusion Retinoic acid can regulate the expression of Tbx1 gene during the development of zebrafish, and this regulation has a significant dose-dependent manner. Retinoic acid is a major non-genetic factor that leads to aberrant Tbx1 expression during embryonic development and may further lead to pharyngeal arch and cardiac dysplasia.