2012-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/643525摘要：基質金屬蛋白酵素為一個蛋白質家族，在哺乳類動物的主要功能為參與組織改造；如胚胎成長、發育，生殖過程以及傷口癒合。此外，這些內生性酵素的降解產物也和先天性免疫反應的啟動有關。至今，已經在人類體內發現超過二十個基質金屬蛋白酵素，而在果蠅也發現兩個相對應的基質金屬蛋白酵素。雖然基質金屬蛋白酵素家族在哺乳類動物以及果蠅有許多詳細的研究，然而，我們對於蚊子的基質金屬蛋白酵素家族所扮演的角色所知甚少。最近，我們在埃及斑蚊(登革熱的主要病媒蚊)體內發現六個基質金屬蛋白酵素，這些酵素皆具有基質金屬蛋白酵素所特有的高度保留啟動區域。我們的初步的實驗結果發現，埃及斑蚊基質金屬蛋白酵素可能在對抗病原入侵及調控產卵機制扮演重要角色。因此，本三年期研究計畫擬採用本實驗室已建立之逆向基因操作系統，藉由特殊建構之雙股RNA抑制特定基因的表現，再分析下游產物的表現或生長發育情形，來釐清埃及斑蚊特定之基質金屬蛋白酵素參與發育、產卵及調控先天性免疫所扮演的角色及功能。本研究結果將有助於我們對此蛋白質家族在不同物種演化保留的全面性瞭解，並且對於未來以遺傳操縱來從事病媒控制工作，以及開發新的除蟲藥劑將有相當大的幫助。<br> Abstract: Matrix Metalloproteinases (MMPs) are a family of enzymes that play major roles in tissue remodeling, such as embryonic growth, development, reproductive process and wound healing in mammals. In addition, deregulation production of these endopeptidases is associated with a number of pathological conditions resulting in the activation of innate immune responses. To date, more than 20 human MMPs have been identified and characterized and only two homologues are identified in the fruit fly Drosophila melanogaster. In spite of their intensive investigation in mammals and fruit fly, functional roles of MMP family in the mosquito are largely unknown. Recently, we identified six MMPs in the mosquito Aedes aegypti, the major vector of Dengue virus. All of these MMPs contain a highly conserved catalytic domain and show signal peptide at the N-terminal part. Our preliminary results suggested that mosquito MMPs may play important roles in regulation of vitellogenesis and innate immune response against pathogens. In this three-year proposal, we will apply the well established reverse genetic approach by silencing specific MMPs in the mosquitoes. Silence of different MMPs will be performed by microinjection of double-stranded RNA targeted to specific MMPs or in combination with introducing the inhibitors of MMPs into the mosquitoes. The roles of MMPs in the innate immune responses, vitellogenesis and development will be explored by immune challenging of specific MMP-silenced mosquitoes, oviposition assay and scanning electron microscopy, respectively. Regulatory machinery of MMP in the innate immune response and vitellogenesis or potential crosstalk with other signal regulatory pathways will be examined. Detail characterization of mosquito MMPs in this proposal will provide new insights into the evolutionary conserved features of MMPs. Information gathered in this study will pave the way toward the establishment of efficient strategies for vector control using molecular engineering approaches and development of novel insecticide.埃及斑蚊雙股RNA卵巢發育訊息傳導路徑Aedes aegyptidouble-stranded RNAovary developmentsignal transduction