Visualization and live imaging analysis of a mosquito saliva protein in host animal skin using a transgenic mosquito with a secreted luciferase reporter system

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 pro...

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Bibliographic Details
Published in:Insect molecular biology 2013-12, Vol.22 (6), p.685-693
Main Authors: Yamamoto, D. S., Yokomine, T., Sumitani, M., Yagi, K., Matsuoka, H., Yoshida, S.
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
Anopheles - genetics
Anopheles - physiology
Anopheles stephensi
anopheline antiplatelet protein
live imaging
luciferase
Luciferases
Luminescent Proteins
Mice
mosquito saliva
Optical Imaging - methods
Saliva - chemistry
Salivary Glands - chemistry
Skin - chemistry
Time-Lapse Imaging
transgenic mosquito
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Summary: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 4 h 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.
ISSN:0962-1075
1365-2583
DOI:10.1111/imb.12055