Immunocytochemical Study on Prophenoloxidases of Mosquito Armigeres subalbatus

• Main Authors: Lin, Li-Chun, 林俐均
• Other Authors: Chen, Cgeng-Chen
• Format: Others
• Language: zh-TW
• Published: 2000
• Online Access: http://ndltd.ncl.edu.tw/handle/54458825280152451830

碩士 === 國立陽明大學 === 寄生蟲學研究所 === 88 === Prophenoloxidase (proPO) is a copper-binding enzyme involving in many physiological functions of insects, i.e. cuticular sclerotization, egg tanning, wound healing and melanization of parasites. In this study, we produced 2 polyclonal antibodies against propeptide and copper-binding region of proPO of mosquito Armigeres subalbatus by Esherichia coli expression system.. Using anti-proPO-propeptide polyclonal antibodies, 5 cross-reactive peptides, M.W. 73, 75, 77, 78 and 79 kDa, were identified from mosquito A. subalbatus hemolymph collected with protease inhibitors. On the contrary, no cross-reactivity was observed in mosquito hemolymph collected without protease inhibitors. The amount of 5 cross-reactive peptides was significantly increased following microfilariae (mf) and Hank Balanced Salt Solution (HBSS)-inoculated mosquitoes. The amount of 3 cross-reactive peptides (M.W. 73, 75 and 77 kDa) was increased in the hemolymph of blood-fed mosquitoes as compared to those of normal mosquitoes. Using anti-proPO-copper polyclonal antibody, 2 cross-reactive peptides, M.W. 73 and 75 kDa, were observed from mosquito hemolymph collected with protease inhibitors. On the contrary, 2 smaller peptides, M.W. 68 and 70 kDa, were observed in mosquito hemolymph collected without protease inhibitors. The amount of 73 and 75 kDa peptides in mosquito hemolymph collected with protease inhibitors was significantly increased in HBSS and mf-inoculated mosquitoes. The amount of 68 and 70 kDa peptides in mosquito hemolymph collected without protease inhibitors was increased in mf-inoculated mosquitoes. The results of present study demostrated that 5 proPOs existed in mosquito A. subalbatus hemolymph. The 77 kDa peptide was significantly increased in blood-fed mosquitoes, indicating that 77 kDa may involve in the physiological functions of egg tanning. In the hemolymph of HBSS and mf-inoculated mosquitoes, the 5 cross-reactive peptides were significantly increased, indicating that they may involve in the wound-healing and melanization of microfilariae.