【摘要】 为研究不同光周期条件下转化生长因子-β超家族/Samd (TGF-β/Smad)信号通路在斑马鱼(Danio rerio)卵巢中的作用,利用荧光定量PCR技术检测连续黑暗(0 L∶24 D, DD)、自然光照(14 L∶10 D, LD)和连续光照(24 L∶0 D, LL)条件下斑马鱼卵巢中TGF-β/Smad信号通路中相关基因的相对表达量,并利用免疫组化技术检测不同光周期条件下p-Smad2在斑马鱼卵巢中的定位情况。结果显示,在不同光周期条件下,TGF-β/Smad信号通路中信号分子在斑马鱼卵巢中的表达模式不尽相同,其中配体(tgfb3)、受体(tgfbr2a,tgfbr2b,tgfbr1b)和下游蛋白激酶(smad2和smad3a)的表达趋势一致,即在不同光周期处理3 d后,上述6个基因的相对表达量DD组最高,LL组最低,但处理7 d后则呈相反趋势。在斑马鱼卵黄发生前期至充分生长未成熟期的卵母细胞中均检测到p-Smad2蛋白信号,但光周期处理后未影响p-Samd2在斑马鱼卵巢中的定位。结果表明,光周期变化可能通过改变tgfb3/受体/蛋白激酶基因的表达模式影响斑马鱼的卵巢发育。更多还原

【Abstract】 To study the role of TGF-β/Smad signaling pathway in zebrafish(Danio rerio) ovary under different photoperiod conditions, we detected the relative expression of relevant genes in TGF-β/Smad signaling pathway of zebrafish ovary under continuous dark(0 L∶24 D, DD), natural photoperiod(14 L∶10 D, LD) and continuous light(24 L∶0 D, LL) using quantitative real-time PCR. The location of p-Smad2 in zebrafish ovary was detected by immunohistochemical technology. The results show that the expression patterns of genes involved in TGF-β/Smad signaling pathway were different in zebrafish ovary under different photoperiod conditions, and the expression profiles of ligand(tgfb3), receptor(tgfbr2 a, tgfbr2 b and tgfbr1 b) and downstream protein kinase(smad2 and smad3 a) were consistent. The relative expression levels of the above six genes were the highest in DD group and the lowest in LL group after 3-day treatment of different photoperiods, and an opposite trend was observed after 7 d. The p-Smad2 protein signal was detected in the development of zebrafish oocytes from previtellogenic stage(PV) to full-grown immature stage(FG).Photoperiod treatment did not affect the localization of p-Samd2 in zebrafish ovary. The results suggest that change of photoperiod might affect the ovarian development of zebrafish by changing the expression pattern of genes of tgfb3/receptor/protein kinase.更多还原