模拟微重力下脊椎动物胚胎发育研究模型建立

发布时间：2018-03-01 02:20

  本文关键词： 模拟微重力 胚胎发育 斑马鱼 蛋白质组学　出处：《大连海事大学》2010年硕士论文　论文类型：学位论文

【摘要】： 太空微重力环境会引起人体多种生理和病理现象,如心血管功能障碍、骨丢失、免疫功能下降、肌肉萎缩、内分泌机能紊乱等,其作用机制一直是空间生物学效应研究中重要的科学问题之一。太空搭载受飞行次数、生保安全和携带体积／重量等诸多因素的限制,且与辐射等作用同时存在,严重限制了微重力生物学效应的研究,因此急需建立地基水平的模拟微重力效应研究平台。本论文从微重力对胚胎发育影响的全新角度出发,建立模拟微重力条件下脊椎动物胚胎发育研究的新模型,为揭示微重力的生物学效应提供帮助。 本文以斑马鱼(Danio rerio)胚胎作为地面模拟微重力效应的研究模型,采用美国航天航空局(NASA)研制的微重力模拟系统(Rotary Cell Culture System, RCCS),对处于器官系统发育关键时期的斑马鱼胚胎进行模拟微重力处理。并应用蛋白质双向电泳(2D-PAGE)技术,对微重力处理前后斑马鱼胚胎的蛋白表达谱进行分析。在获取的28个差异性表达(p≤0.05)的蛋白点中,共选取10个蛋白点进行MALDI-TOF/TOF质谱测序。生物信息学初步分析表明,这些差异表达的蛋白质与细胞结构、肌肉组成及能量代谢等生理活动密切相关,部分蛋白质在已有的微重力研究中未被报道过。上述结果表明,本论文建立的模拟微重力脊椎动物胚胎发育研究模型,确实可以为揭示微重力的生物学效应提供新颖的目标分子。 为了进一步探讨模拟微重力对蛋白质表达调控的影响,本文随机选取了四个测序获得的差异表达蛋白(肌肉型的肌酸激酶b,β-肌动蛋白2,135kDa中心体蛋白,原肌球蛋白4)以及与细胞损伤修复和凋亡相关的p53蛋白作为目标分子,应用实时荧光定量PCR(RT-qRCR)技术,对上述目标分子在模拟微重力作用下的mRNA表达水平进行了相对定量分析,获得了上述目标分子产生模拟微重力效应的响应时间。结果表明,在不同的发育阶段对斑马鱼胚胎进行不同时间的模拟微重力处理,基因的表达水平会产生不同的变化趋势。
[Abstract]:The microgravity environment in space will cause many physiological and pathological phenomena, such as cardiovascular dysfunction, bone loss, decreased immune function, muscular atrophy, endocrine dysfunction, etc. The mechanism of its action is always one of the important scientific problems in the study of space biological effects. Space carrier is restricted by many factors, such as flight times, safety of life and carrying volume / weight, and exists simultaneously with radiation and so on. The study of biological effects of microgravity is seriously restricted, so it is urgent to establish a platform for the study of simulated microgravity effects at the foundation level. In this paper, the effects of microgravity on embryonic development are viewed from a new perspective. A new model for the study of vertebrate embryonic development under simulated microgravity is established to help reveal the biological effects of microgravity. In this paper, the zebrafish Danio rerio embryo was used as the ground simulation model of microgravity effect. A microgravity simulation system, Rotary Cell Culture system, developed by NASA, was used to simulate microgravity treatment of zebrafish embryos at the critical stage of organ phylogenetic development. Two-dimensional protein electrophoresis (2D-PAGE) technique was used to treat zebrafish embryos. The protein expression profiles of zebrafish embryos before and after microgravity treatment were analyzed. Among the 28 protein sites with differential expression p 鈮，

本文编号：1549985