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    Title: Vha16-1對果蠅腸道功能和壽命之調控
    Vha16-1 regulates intestinal function and lifespan in Drosophila melanogaster
    Authors: 宋祐陞
    Contributors: 王培育
    宋祐陞
    Keywords: 突變生成
    壽命
    肥胖
    腸道酸化
    mutagenesis
    lifespan
    obesity
    gut acidification
    Date: 2011
    Issue Date: 2012-10-30 15:22:11 (UTC+8)
    Abstract: 突變生成(mutagenesis)的方式有許多種，其中insertional mutagenesis為果蠅上常使用建立突變株的方式，本篇論文利用p[GawB]隨機插入果蠅genome中產生大量突變株，並篩選出會影響壽命的突變果蠅M2。進一步的實驗發現M2果蠅為Vha16-1基因的突變，並造成其mRNA表現量的下降，且在低卡路里(5% yeast、5% dextrose)與高卡路里(15% yeast、15% dextrose)的環境下homozygous mutant果蠅皆有減少平均壽命的現象。 Vha16-1所表現的蛋白為Vacuolar-type H+-ATPase (V-ATPase)上的subunit c，V-ATPase主要的功能為藉由消耗ATP來運送氫離子，並可調節胞器或胞外腔室的酸鹼平衡。V-ATPase主要表現在果蠅腸道的copper cell上，此細胞的功能類似於哺乳動物的胃壁細胞(parietal cells)，與胃酸的分泌有關，我們發現M2 homozygous mutant果蠅因Vha16-1基因的缺失而有減少腸道酸化的情形發生，符合我們觀察到其在腸道上的表現。此一現象亦在另一株突變果蠅Vha16-1EP2372上加以證實。先前研究顯示果蠅腸道酸鹼平衡的破壞會影響到對養分的吸收，而Vha16-1的缺失亦導致M2果蠅體重與三酸甘油酯的上升，並增加對飢餓的耐受性，而這些代謝上的變化並不會改變M2果蠅對食物的攝取量或者生育能力。綜合這些實驗結果，我們推測Vha16-1基因的缺失會改變腸道功能，並影響果蠅體內代謝的狀態，表現出類似肥胖(obesity)的性狀，而終導致平均壽命的縮短。
    Mutagenesis can be induced by many ways and one of the most common approaches used in Drosophila is insertional mutagenesis. In this study, we screened pGawB insertion lines and identified M2 as a novel mutant with affected lifespan. The mutant allele of M2 carried a pGawB inseration at the 5’ end of the Vha16-1 gene, which caused a reduced Vha16-1 mRNA expression level and a shorten lifespan in homozygous mutants under both low calorie (5% yeast and 5% dextrose) and high calorie (15% yeast and 15% dextrose) foods. Vha16-1 encodes the c subunit of the Vacuolar-type H+-ATPase (V-ATPase) which is known to regulate pH homeostasis by pumping protons across organelle and plasma membranes. V-ATPase is highly expressed by the Copper cells which are located at the Drosophila middle midgut and functionally similar to the gastric acid producing parietal cells in mammals. Along the same line, we found that Vha16-1 pGawB drives GFP reporter was observed along the Drosophila gastrointestinal tract. M2 as well as the other Vha16-1 hypomorphic mutant line, EP2372, also showed reduced midgut acidification. This disrupted pH homeostasis in the Drosophila midgut region may be associated with increased body weight, triglyceride, and starvation resistance that observed in M2 mutants. The feeding behavior and reproductive function, however, were not affected in M2 mutant flies. In summary, our data suggested Vha16-1 deficits may alter normal intestinal function or internal metabolic status that ultimately induces obesity phenotypes with reduced lifespan.
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    Description: 碩士
    國立政治大學
    神經科學研究所
    98754008
    100
    Source URI: http://thesis.lib.nccu.edu.tw/record/#G0098754008
    Data Type: thesis
    Appears in Collections:[Graduate Institute of Neuroscience] Theses

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