本文關(guān)鍵詞： 模擬微重力 胚胎發(fā)育 斑馬魚 蛋白質(zhì)組學(xué)　出處：《大連海事大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 太空微重力環(huán)境會引起人體多種生理和病理現(xiàn)象,如心血管功能障礙、骨丟失、免疫功能下降、肌肉萎縮、內(nèi)分泌機能紊亂等,其作用機制一直是空間生物學(xué)效應(yīng)研究中重要的科學(xué)問題之一。太空搭載受飛行次數(shù)、生保安全和攜帶體積／重量等諸多因素的限制,且與輻射等作用同時存在,嚴重限制了微重力生物學(xué)效應(yīng)的研究,因此急需建立地基水平的模擬微重力效應(yīng)研究平臺。本論文從微重力對胚胎發(fā)育影響的全新角度出發(fā),建立模擬微重力條件下脊椎動物胚胎發(fā)育研究的新模型,為揭示微重力的生物學(xué)效應(yīng)提供幫助。 本文以斑馬魚(Danio rerio)胚胎作為地面模擬微重力效應(yīng)的研究模型,采用美國航天航空局(NASA)研制的微重力模擬系統(tǒng)(Rotary Cell Culture System, RCCS),對處于器官系統(tǒng)發(fā)育關(guān)鍵時期的斑馬魚胚胎進行模擬微重力處理。并應(yīng)用蛋白質(zhì)雙向電泳(2D-PAGE)技術(shù),對微重力處理前后斑馬魚胚胎的蛋白表達譜進行分析。在獲取的28個差異性表達(p≤0.05)的蛋白點中,共選取10個蛋白點進行MALDI-TOF/TOF質(zhì)譜測序。生物信息學(xué)初步分析表明,這些差異表達的蛋白質(zhì)與細胞結(jié)構(gòu)、肌肉組成及能量代謝等生理活動密切相關(guān),部分蛋白質(zhì)在已有的微重力研究中未被報道過。上述結(jié)果表明,本論文建立的模擬微重力脊椎動物胚胎發(fā)育研究模型,確實可以為揭示微重力的生物學(xué)效應(yīng)提供新穎的目標分子。 為了進一步探討模擬微重力對蛋白質(zhì)表達調(diào)控的影響,本文隨機選取了四個測序獲得的差異表達蛋白(肌肉型的肌酸激酶b,β-肌動蛋白2,135kDa中心體蛋白,原肌球蛋白4)以及與細胞損傷修復(fù)和凋亡相關(guān)的p53蛋白作為目標分子,應(yīng)用實時熒光定量PCR(RT-qRCR)技術(shù),對上述目標分子在模擬微重力作用下的mRNA表達水平進行了相對定量分析,獲得了上述目標分子產(chǎn)生模擬微重力效應(yīng)的響應(yīng)時間。結(jié)果表明,在不同的發(fā)育階段對斑馬魚胚胎進行不同時間的模擬微重力處理,基因的表達水平會產(chǎn)生不同的變化趨勢。
[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，

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