- 期刊
Fluorescence Immunohistochemistry for Monitoring the Transmission Pathway of Aedes aegypti (L.) Male Accessory Gland MAG-317 Protein during Copulation
以螢光免疫化學法追蹤雄埃及斑蚊副腺蛋白MAG-317在交尾時的傳導途徑
摘要
目的：登革熱是Togaviridae族群的病毒，在亞熱帶地區每年都會經病媒蚊傳播斷骨熱及登革出血熱，尤其是在台灣，本研究主要探討埃及斑蚊雄蚊副腺蛋白MAG-317，經由交媾時傳輸至雌蚊貯精囊的MAG-317蛋白追蹤探討。 方法：擷取電泳分析後粗埃及雄蚊副腺蛋白質中的MAG-317純蛋白，研製特異性多株抗體（MAG-317特異性多株抗體），用來偵測追蹤MAG-317純蛋白經由交尾後在雌蚊體內各組織之分佈。本研究室整合登革熱感染的模式，由雌蚊注射的血液採檢、經由蚊子細胞株培養，再經由結合螢光免疫的血清型特異性單株抗體證實。 結果：本研究研發出螢光免疫定位法定量追蹤雄蚊副腺蛋白走向。螢光抗體與雄蚊副腺蛋白結合後，經由交尾注射入雌蚊體內，螢光抗體反應可在雌蚊的貯精囊，卵巢，輸卵管，已受精卵和子代中發現。 結論：本研究證實埃及斑蚊之雄蚊副腺蛋白MAG-317可經由交尾途徑傳輸至雌蚊貯精囊。
並列摘要
Purpose: The male accessory gland (MAG) protein modulates the host-seeking behavior and stimulating oviposition of female Aedes aegypti (L.) mosquitoes. In this study, a venereal transmission of MAG-317 from the mosquito Aedes aegypti (L.) into spermathecae of gravid females was tracked unambiguously during copulation. Methods: The specificity of the polyclonal antibodies rose against the gel-purified MAG-317 protein from MAGS of Ae. aegypti. This antiserum was subsequently used for the detection of MAG-317 protein in various tissues. Fifty three-day-old unmated labeled male mosquito's were fed an F solution (FITC-conjugated secondary Ab/glucose solution) for two days and placed into a rearing box with 25 three-day-old virgin females for random copulation. The labeled males and mated females were dissected and the accessory glands or spermathecae of each individual were examined using a confocal laser-scanning microscope. Results: In this study, fluorescein was incorporated into target protein in MAG and, fluorescein reaction in inseminated females was subsequently detected. The protein was found to be transmitted during copulation, and fluorescein reaction was seen in spermathecae, ovaries, oviducts, fertile eggs, and offspring. Conclusion: The MAG-317 protein from Aedes aegypti (L.) is transmitted into spermathecae of gravid females during copulation.
延伸閱讀
- 蘇兪丞、徐爾烈、何兆美（2003）。運用隨機增幅多型性DNA聚合酶連鎖反應技術區別埃及斑蚊（Diptera: Culicidae）腹部背板白鱗分佈的差異。台灣昆蟲，23(3)，vi+225-237。https://doi.org/10.6661/TESFE.2003021
- Cheang, K. W. (2018). 探討在登革病毒感染下埃及斑蚊之唾液/SRPN23唾液蛋白對免疫反應與出血反應所造成的影響 [master's thesis, National Taiwan University]. Airiti Library. https://doi.org/10.6342/NTU201803692
- 翁仕哲（2020）。埃及斑蚊生殖調控路徑及宿主因子調控登革病毒之複製〔博士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU202000529
- 吳秉錕（2017）。探討未折疊蛋白質反應誘發之自噬作用參與埃及斑蚊體內登革病毒之複製〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU201702828
- Lee, Y. C. (2022). 探討埃及斑蚊唾液蛋白對登革病毒複製與病媒蚊攝食行為之影響 [master's thesis, National Taiwan University]. Airiti Library. https://doi.org/10.6342/NTU202202717