双酚AF胁迫对斑马鱼早期发育相关基因的影响

Effects of BPAF on the expression of early development genes of zebrafish(Danio rerio)embryo and larvae

[目的]探讨双酚AF胁迫对斑马鱼早期发育的影响。[方法]采用半静态染毒的方式暴露受精卵7d(0、5、50和500μg/L),通过显微观察与实时荧光定量PCR方法,检测孵化率及相关基因表达量变化。[结果]50、500μg/L BPAF暴露影响斑马鱼胚胎72 h孵化率,分别下降44.97%、49.44%;下丘脑trh基因在各暴露组无明显变化,而下丘脑crh基因在胚胎(幼鱼)期各暴露组分别上调了44.14%(40.45%)、38.55%(95.17%)、70.65%(83.25%);胚胎期垂体tsh-β基因表达水平在50、500μg/L浓度组显著上调,而幼鱼期tsh-β基因表达水平仅在50μg/L暴露组显著上调。[结论]双酚AF胁迫能够降低斑马鱼胚胎72 h孵化率,crh基因对BPAF胁迫较敏感,最低有影响浓度LOEC为5μg/L。

[Objective]To investigate the effects and mechanism of low doses of BPAF on the development of zebrafish embryos and larvae. [Methods] The fertilized egg were exposed to 5,50 and 500 μg/L BPAF for 7 days based on the semi static method. Through microscopic observation,we studied the effects of BPAF on the early development of zebrafish embryos（2 days） and the larvae（7 days）. We detected effects on the expression of related genes by qRT-PCR. [Results] The hatching rate of zebrafish embryos decreased by 44. 97% and 49. 44% at 72 hours in 50 and 500 μg/L BPAF treatments,respectively. The expression of trh mRNA of hypothalamus had no significant change in different exposed groups. The expression of crh mRNA of hypothalamus was up-regulated in embryos and by 44. 14%,38. 55% and 70. 65% in different concentration BPAF（5,50 and 500μg/L）. Meanwhile,the expression of crh mRNA in the hypothalamus was increased in larvae by44. 14%,38. 55% and 70. 65% in the different exposure groups. The expression of tsh-β mRNA in zebrafish embryos was significantly up-regulated by 50 and 500 μg/L BPAF. The expression of tsh-β mRNA in the larvae was significantly increased by 50 μg/L BPAF. [Conclusion] BPAF could reduce the hatching rate of zebrafish embryos at 72 hours. The crh gene was more sensitive to BPAF exposure. The LOEC was 5 μg/L.