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Study of Hypoxia on Embryogenesis, Pharmaceutical Testing and Stem Cell Regulation Using Drosophila Model

機(jī)譯：利用果蠅模型研究缺氧對胚胎發(fā)生，藥物測試和干細(xì)胞調(diào)控的影響

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Drosophila melanogaster has been used to study human disease as a model organism for many years. Many basic biological, physiological, and neurological properties are conserved between mammals and fly. We investigated its applications in the study of the impact of environmental stress on embryogenesis and its compensating mechanism, its uniqueness and powerfulness in modern pharmaceutical testing and screening, and its application in gene function identification.;First we directly investigated Drosophila embryo development in vivo inside a customized microfluidic device with an established local oxygen gradient on a micrometer scale. When the embryos were placed in various conditions, two of the key developmental activities, the germ band shortening and the tail retraction, were examined during the embryogenesis. The time-lapse live cell imaging technique was used to monitor the cell morphology changes and pattern migration with the help of green fluorescence protein markers. Our results show that the examined activities during the Drosophila embryogenesis are highly sensitive to oxygen concentrations. Using this information, we presented a model to estimate the oxygen permeability across the Drosophila embryonic layers for the first time. Secondly, the Drosophila testis was used to evaluate the basic therapeutic mechanisms of the active components of several traditional Chinese medicines (TCM) that is known related to animal genital system and sexual function. Specifically, we investigated the effect of the compounds that were extracted from above-mentioned Chinese medicines on Drosophila germline stem cells (GSCs) by quantifying the GSCs mitotic activity and GSC number. Our results showed that, flies have a significantly higher cell cycle index when fed at certain concentration of icariin and Tanshinone IIA, the primary active component of YYH and DS, respectively. Other tested concentrations of extract produced either toxicity or insignificant effects on the mitotic activity. This indicates their function of promoting the GSCs mitosis. At last, we analyzed the expression and localization of two polarity genes throughout the cell cycle, and investigated how they affected mitotic spindle dynamics in asymmetric stem cell divisions. In stem cell divisions, it is critical to maintain tissue homeostasis by balancing the number of stem cells and progenitor cells. Improperly balancing may result in tumorigenesis due to tissue hyper-proliferation or tissue ageing due to tissue degeneration. Previous studies show that cyst stem cells (CySCs) in Drosophila testis divide asymmetrically. This behavior is ensured by the stem cell mitotic spindle repositioning, during which one of the spindle poles always moves close to the stem cell niche (a.k.a. hub cells) near the onsite of anaphase. Known as polarity proteins, the apically localized Par polarity complex, containing Bazooka (Baz; homolog of Par-3 in D. melanogaster), its binding target atypical Protein Kinase C (aPKC), and Par-6, are widely reported to be crucial in polarized cell epithelium and asymmetric cell division in multiple stem cell systems. We found that Baz and aPKC are required in Drosophila CySC asymmetric cell division.
機(jī)譯：果蠅（Drosophila melanogaster）已作為模型生物被用于研究人類疾病多年。哺乳動(dòng)物和果蠅之間保留了許多基本的生物學(xué)，生理和神經(jīng)學(xué)特性。我們研究了其在研究環(huán)境脅迫對胚胎發(fā)生及其補(bǔ)償機(jī)制的影響中的應(yīng)用，在現(xiàn)代藥物測試和篩選中的獨(dú)特性和強(qiáng)大性，以及在基因功能鑒定中的應(yīng)用。；首先，我們直接研究了果蠅體內(nèi)的胚胎發(fā)育定制的微流控設(shè)備，該設(shè)備具有已建立的微米級(jí)局部氧梯度。當(dāng)胚胎處于各種條件下時(shí)，在胚胎發(fā)生過程中檢查了兩個(gè)關(guān)鍵的發(fā)育活動(dòng)，即胚帶縮短和尾巴回縮。延時(shí)活細(xì)胞成像技術(shù)用于借助綠色熒光蛋白標(biāo)記物監(jiān)測細(xì)胞形態(tài)變化和模式遷移。我們的結(jié)果表明，在果蠅胚胎發(fā)生過程中檢查的活性對氧濃度高度敏感。利用這些信息，我們首次提出了一個(gè)模型來估計(jì)果蠅胚胎各層的透氧性。其次，使用果蠅睪丸評估了幾種與動(dòng)物生殖系統(tǒng)和性功能有關(guān)的中藥（TCM）有效成分的基本治療機(jī)制。具體而言，我們通過量化GSC的有絲分裂活性和GSC數(shù)量，研究了從上述中藥中提取的化合物對果蠅種系干細(xì)胞（GSC）的影響。我們的研究結(jié)果表明，當(dāng)飼喂一定濃度的icariin和Tanshinone IIA（分別為YYH和DS的主要活性成分）時(shí)，果蠅具有明顯更高的細(xì)胞周期指數(shù)。其他測試濃度的提取物對有絲分裂活性產(chǎn)生毒性或微不足道的影響。這表明它們具有促進(jìn)GSCs有絲分裂的功能。最后，我們分析了兩個(gè)極性基因在整個(gè)細(xì)胞周期中的表達(dá)和定位，并研究了它們?nèi)绾斡绊懖粚ΨQ干細(xì)胞分裂中的有絲分裂紡錘體動(dòng)力學(xué)。在干細(xì)胞分裂中，通過平衡干細(xì)胞和祖細(xì)胞的數(shù)量來維持組織穩(wěn)態(tài)是至關(guān)重要的。平衡不當(dāng)可能會(huì)由于組織過度增殖或由于組織變性而導(dǎo)致組織老化而導(dǎo)致腫瘤發(fā)生。先前的研究表明，果蠅睪丸中的囊性干細(xì)胞（CySCs）不對稱分裂。干細(xì)胞有絲分裂紡錘體重新定位可確保這種行為，在此過程中，紡錘極之一始終移至后期后期附近的干細(xì)胞小生境（又稱集線器細(xì)胞）附近。廣為人知的是，被稱為極性蛋白的根尖定位的Par極性復(fù)合物（包括火箭筒（Baz；黑腹果蠅中的Par-3的同系物），其結(jié)合靶標(biāo)非典型蛋白激酶C（aPKC）和Par-6）是至關(guān)重要的。在多個(gè)干細(xì)胞系統(tǒng)中極化細(xì)胞上皮細(xì)胞和不對稱細(xì)胞分裂。我們發(fā)現(xiàn)，果蠅CySC不對稱細(xì)胞分裂中需要Baz和aPKC。

著錄項(xiàng)

  • 作者

    Wang, Zhinan.;

  • 作者單位

    University of Illinois at Chicago.;

  • 授予單位 University of Illinois at Chicago.;
  • 學(xué)科 Biomedical engineering.
  • 學(xué)位 Ph.D.
  • 年度 2017
  • 頁碼 123 p.
  • 總頁數(shù) 123
  • 原文格式 PDF
  • 正文語種 eng
  • 中圖分類 遙感技術(shù);
  • 關(guān)鍵詞

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