The effects of collagen genes col-131, col-150, dpy-5 and col-121 on fertility and development of Caenorhabditis elegans
首发时间:2015-10-16
Abstract:5-Fluorouracil (5-FU) treatment in Caenorhabditis elegans(C.elegans) leads to slow developmental rate, reduces brood size, small body size, and truncates lifespan. Using Digital Gene Expression (DGE) profiling analysis, the potential pathways and genes that could be involved in declining the embryos of C. elegans have identified. Here, when C.elegans treated by 5-FU, three down-regulated genes were identified, including col-131, col-150 and dpy-5, which belong to ECM-receptor interaction pathway and Focal adhesion and one up-regulated gene col-121 which acts in Focal adhesion. Furtherly, RNAi-mediated gene knock down method was used to examine their RNAi phenotypes. The result showed that dpy-5(RNAi) worms were short-body, dumpy, and displaying a serious defect in fertility. col-121(RNAi) worms were slender, long, and slightly altered in body development. In addition, col-121(RNAi) worms displayed abnormal body morphology. Furthermore, knocking down of col-131 has subtle side effect in C.elegans fertility; however, col-131 (RNAi) worms could rescue 5-Fu toxic effect on C.elegans on fertility and lifespan defect. Taken together, the results revealed the essential role for col-150, dpy-5 and col-121 in C.elegans fertility decline. Our data also suggested that col-131 may be an essential gene in ECM-receptor interaction pathway and Focal adhesion and regulate other cuticle collagen genes including col-150, dpy-5, and col-121.
keywords: Caenorhabditis elegans, collagen genes, 5-FU, RNAi
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胶原蛋白基因col-131、col-150、col-121和dpy-5对秀丽隐杆线虫生殖与发育的影响
摘要:目的:本论文通过分析5-氟尿嘧啶作用秀丽隐杆线虫以后基因的表达情况,利用RNA干扰技术研究基因的沉默表达对线虫生殖和寿命的影响,分析胶原蛋白家族基因在秀丽隐杆线虫生殖及其他生命活动中的重要作用,讨论5-氟尿嘧啶对线虫的作用机制。方法:利用高通量数字基因表达谱分析,筛选5-氟尿嘧啶作用后基因表达上调或者下调十分显著的基因,利用RNA干扰技术得到基因沉默表达型线虫,利用光学显微镜观察RNA干扰型线虫形态、产卵量及寿命变化,分析基因的功能及其在通路中的作用。结果:利用数字基因表达谱,筛选得到在5-氟尿嘧啶的处理下表达下调显著的col-131、col-150和dpy-5基因和表达上调显著的col-121基因;col-131、col-150和dpy-5基因属于ECM-receptor和Focal adhesion通路;col-121基因属于Focal adhesion通路。利用饲喂法,将人工构建的RNA干扰细菌喂食秀丽隐杆线虫,col-131、col-150、col-121和dpy-5基因表达量均显著降低,得到RNA干扰型线虫。dpy-5基因RNA干扰型线虫身体短粗,表现出严重的生殖缺陷;col-121基因RNA干扰型线虫细长,生殖与发育受到影响;col-150基因RNA干扰型线虫产卵量和生存率都有所降低。col-131基因表达沉默对秀丽隐杆线虫生殖和寿命没有明显影响,但是col-131基因沉默可以减弱5-氟尿嘧啶对秀丽隐杆线虫生殖的毒性。col-131基因RNA干扰型线虫可以在含有5-氟尿嘧啶的NGM培养基上产卵。当线虫col-131基因表达沉默以后,线虫的col-150、col-121和dpy-5基因表达量均降低。本论文结果表明,col-150、col-121和dpy-5基因对秀丽隐杆线虫生殖的调节起到重要的作用。col-131基因可能是与5-氟尿嘧啶作用的靶点密切相关的基因,并且在ECM-receptor interaction通路和 Focal adhesion通路中调节其他胶原蛋白家族基因的表达。
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胶原蛋白基因col-131、col-150、col-121和dpy-5对秀丽隐杆线虫生殖与发育的影响
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