SP600125孵育影响鲫骨骼发育基因表达

前期研究表明ATP竞争性小分子抑制剂SP600125在体外可以高效诱导鱼类细胞多倍化。研究首次尝试用SP600125直接孵育鲫受精卵以获得多倍体鱼的可能性。结果显示SP600125处理受精卵虽然能够引起部分胚胎倍性发生改变, 但也显著影响胚胎发育并导致胚胎高死亡率, 同时伴随着畸形鱼苗的发生, 其中主要表现为脊椎弯曲和尾缺失等骨骼发育异常。SP600125处理导致骨骼畸形在SP600125孵育鲫鱼苗试验中也同样获得验证。荧光定量PCR检测结果表明, 在体(受精卵和鱼苗)或离体(培养的尾鳍细胞)状态下, SP600125孵育会引起smad6、stat1、 lef1、bmp2和twist1等骨骼发育相关基因mRNA水平显著上调。相关结果为进一步调整SP600125诱导的鱼类倍性操作策略和方法提供了重要理论参考。

THE EFFECT OF SP600125 ON THE EXPRESSION OF SKELETAL GENES OF CRUCIAN CARP(CARASSIUS AURATUS VAR)

It has been shown that the ATP competitive small molecule inhibitor SP600125 can efficiently induce fish cell polyploidization in vitro.In this study,we used SP600125 to directly incubate the fertilized eggs of crucian carp to obtain the possibility of polyploid fish,and found that SP600125 caused partial embryo ploidy,significantly affected the embryonic development and led to high mortality of embryos.SP600125 also induced a large number of malformed fry after treating fertilized eggs,mainly skeletal dysplasia such as spinal curvature and tail loss.The skeletal malformation caused by SP600125 was further validated.Real-time PCR indicated that SP600125 significantly induced mRNA levels of skeletal development-related genes,such as smad6,stat1,lef1,bmp2 and twist1 in vivo(fertilized eggs and fry)or in vitro(cultured caudal fin cells).These results clearly help to develop a potential mode of fish polyploidy induced by SP600125.