Beerntsen B T, Christensen B M
Department of Veterinary Science, University of Wisconsin, Madison 53706.
Exp Parasitol. 1990 Nov;71(4):406-14. doi: 10.1016/0014-4894(90)90066-l.
[35S]Methionine-labeled hemolymph polypeptides from adult, female Aedes aegypti Liverpool strain mosquitoes inoculated with the microfilariae of the filarial nematode Dirofilaria immitis were compared with those from saline-inoculated and uninoculated controls by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by fluorography. SDS-PAGE analysis of cell-free hemolymph collected via perfusion at 6, 12, 24, 48, 72, and 96 hr postinoculation (PI) detected the enhanced expression of an 84-kDa polypeptide. This polypeptide, expressed constitutively in the hemolymph of all three groups of mosquitoes, increased considerably in inoculated mosquitoes as time progressed as compared with uninoculated controls. Moreover, the 84-kDa polypeptide was expressed at higher levels in D. immitis-inoculated mosquitoes than in saline-inoculated controls. This stimulation of de novo biosynthesis of the 84-kDa polypeptide in inoculated mosquitoes may play a role in the immune response of mosquitoes. Since it is likely that the wound healing response in insects involves many of the same chemical processes as occur in melanotic encapsulation reactions of mosquitoes against filarial worms, the preferential expression of the 84-kDa polypeptide in saline-inoculated mosquitoes seen in this study may reflect a wound healing response. The greater increase in synthesis of this protein in D. immitis-inoculated mosquitoes may reflect production of melanotic material required for parasite destruction as well as for wound healing.
将感染了丝状线虫犬恶丝虫微丝蚴的成年雌性埃及伊蚊利物浦品系蚊子的[35S]甲硫氨酸标记血淋巴多肽，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）及荧光自显影，与接种生理盐水和未接种的对照蚊子的血淋巴多肽进行比较。对接种后6、12、24、48、72和96小时（PI）通过灌注收集的无细胞血淋巴进行SDS-PAGE分析，检测到一种84 kDa多肽的表达增强。这种多肽在三组蚊子的血淋巴中组成性表达，与未接种对照相比，接种蚊子体内随着时间推移其含量显著增加。此外，接种犬恶丝虫的蚊子中84 kDa多肽的表达水平高于接种生理盐水的对照。接种蚊子中84 kDa多肽的从头生物合成受到刺激，这可能在蚊子的免疫反应中起作用。由于昆虫的伤口愈合反应可能涉及许多与蚊子对丝虫的黑化包囊反应相同的化学过程，本研究中在接种生理盐水的蚊子中观察到的84 kDa多肽的优先表达可能反映了伤口愈合反应。接种犬恶丝虫的蚊子中这种蛋白质合成的更大增加可能反映了破坏寄生虫以及伤口愈合所需的黑化物质的产生。
