Visualization and live imaging analysis of a mosquito saliva protein in host animal skin using a transgenic mosquito with a secreted luciferase reporter system
Visualization and live imaging analysis of a mosquito saliva protein in host animal skin using a transgenic mosquito with a secreted luciferase reporter system
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DOI：
10.1111/imb.12055
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发表时间：
2013-12-01
影响因子：
2.6
通讯作者：
Yoshida, S.
中科院分区：
文献类型：
--
作者：
Yamamoto, D. S.;Yokomine, T.;Yoshida, S.
蚊子在吸血时将唾液注入脊椎动物宿主体内。宿主皮肤蚊虫唾液的分析对于阐明蚊虫叮咬后的炎症反应、抗血栓药物的开发以及媒介传播疾病的传播阻断具有重要意义。我们培育了在唾液腺中表达分泌荧光素酶蛋白（MetLuc）与唾液蛋白（AAPP）融合的转基因斯氏按蚊。转基因蚊子的产物（AAPP-MetLuc）既具有作为MetLuc的荧光素酶活性，又具有作为AAPP与胶原的结合活性。转基因蚊子叮咬小鼠皮肤的发光检测表明，AAPP-MetLuc以唾液成分的形式通过喂血注入小鼠皮肤。AAPP-MetLuc在吸血后至少在蚊虫叮咬部位保持荧光素酶活性4h。AAPP也被怀疑存在于蚊子叮咬造成的损伤部位，并通过与胶原蛋白结合来阻止血小板聚集。这些结果证明了活体脊椎动物宿主皮肤蚊虫唾液可视化和延时分析方法的建立。该技术可为蚊子唾液注射到宿主皮肤后的分析提供便利，为开发新的药物和疾病控制策略提供便利。
Mosquitoes inject saliva into a vertebrate host during blood feeding. The analysis of mosquito saliva in host skin is important for the elucidation of the inflammatory responses to mosquito bites, the development of antithrombotic drugs, and the transmission-blocking of vector-borne diseases. We produced transgenic Anopheles stephensi mosquitoes expressing the secretory luciferase protein (MetLuc) fused to a saliva protein (AAPP) in the salivary glands. The transgene product (AAPP-MetLuc) of transgenic mosquitoes exhibited both luciferase activity as a MetLuc and binding activity to collagen as an AAPP. The detection of luminescence in the skin of mice bitten by transgenic mosquitoes showed that AAPP-MetLuc was injected into the skin as a component of saliva via blood feeding. AAPP-MetLuc remained at the mosquito bite site in host skin with luciferase activity for at least 4h after blood feeding. AAPP was also suspected of remaining at the site of injury caused by the mosquito bite and blocking platelet aggregation by binding to collagen. These results demonstrated the establishment of visualization and time-lapse analysis of mosquito saliva in living vertebrate host skin. This technique may facilitate the analysis of mosquito saliva after its injection into host skin, and the development of new drugs and disease control strategies.