The RNA interference pathway affects midgut infection- and escape barriers for Sindbis virus in Aedes aegypti.
The RNA interference pathway affects midgut infection- and escape barriers for Sindbis virus in Aedes aegypti.
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DOI：
10.1186/1471-2180-10-130
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发表时间：
2010-04-28
期刊：
影响因子：
4.2
通讯作者：
Franz AW
中科院分区：
文献类型：
--
作者：
Khoo CC;Piper J;Sanchez-Vargas I;Olson KE;Franz AW
RNA干扰（RNAi）途径作为埃及伊蚊的先天性抗病毒免疫应答，调节蚊子的虫媒病毒感染。辛德毕斯病毒（SINV;披膜病毒科，甲病毒属）是一种感染山羊的虫媒病毒。实验室里的埃及人SINV TR 339株在感染Ae.埃及人。这一障碍的性质尚未得到很好的理解。为了研究中肠作为决定虫媒病毒载体能力的中央器官的作用，我们产生了转基因蚊子，其中RNAi途径在吸血雌性的中肠组织中受损。我们使用这些蚊子来揭示中肠中的RNAi损伤对SINV复制、中肠感染和传播效率以及蚊子寿命的影响。作为研究虫媒病毒-蚊子相互作用的新工具，我们通过沉默Aa-dcr 2基因的表达，设计了一种转基因蚊子系，该蚊子系在吸血雌性的中肠中具有受损的RNAi途径。在转基因Carb/dcr 16系的中肠组织中，与受体蚊子品系相比，在血餐后1-7天（pbm）之间，Aa-dcr 2表达降低约50%。在用SINV-TR 339 EGFP感染后，Aa-dcr 2表达水平在两种蚊子品系中均增强。在RNAi途径中，受损的蚊子品系SINV滴度和中肠感染率在7天pbm时显著更高。在转基因蚊子中，病毒传播率也有增加的强烈趋势。在7-14天pbm之间，SINV在转基因蚊子的中肠组织中减少。RNAi途径的转基因损伤和/或SINV感染并不影响蚊子的寿命。我们表明，RNAi受损的转基因蚊子是一个有用的工具，在分子水平上研究虫媒病毒-蚊子的相互作用。被Ae吞噬后。在埃及伊蚊中，重组SINV-TR 339 EGFP面临MEB和中肠感染屏障（MIB）。中肠中RNAi途径的受损强烈降低了病毒的两个中肠屏障。这证实了Ae.埃及伊蚊调节中肠中SINV的载体能力。RNAi途径通过影响中肠的感染率、感染强度和从中肠到次级组织的传播来充当传入病毒的守门人。
The RNA interference (RNAi) pathway acts as an innate antiviral immune response in Aedes aegypti, modulating arbovirus infection of mosquitoes. Sindbis virus (SINV; family: Togaviridae, genus: Alphavirus) is an arbovirus that infects Ae. aegypti in the laboratory. SINV strain TR339 encounters a midgut escape barrier (MEB) during infection of Ae. aegypti. The nature of this barrier is not well understood. To investigate the role of the midgut as the central organ determining vector competence for arboviruses, we generated transgenic mosquitoes in which the RNAi pathway was impaired in midgut tissue of bloodfed females. We used these mosquitoes to reveal effects of RNAi impairment in the midgut on SINV replication, midgut infection and dissemination efficiencies, and mosquito longevity. As a novel tool for studying arbovirus-mosquito interactions, we engineered a transgenic mosquito line with an impaired RNAi pathway in the midgut of bloodfed females by silencing expression of the Aa-dcr2 gene. In midgut tissue of the transgenic Carb/dcr16 line, Aa-dcr2 expression was reduced ~50% between 1-7 days post-bloodmeal (pbm) when compared to the recipient mosquito strain. After infection with SINV-TR339EGFP, Aa-dcr2 expression levels were enhanced in both mosquito strains. In the RNAi pathway impaired mosquito strain SINV titers and midgut infection rates were significantly higher at 7 days pbm. There was also a strong tendency for increased virus dissemination rates among the transgenic mosquitoes. Between 7-14 days pbm, SINV was diminished in midgut tissue of the transgenic mosquitoes. Transgenic impairment of the RNAi pathway and/or SINV infection did not affect longevity of the mosquitoes. We showed that RNAi impaired transgenic mosquitoes are a useful tool for studying arbovirus-mosquito interactions at the molecular level. Following ingestion by Ae. aegypti, the recombinant SINV-TR339EGFP was confronted with both MEB and a midgut infection barrier (MIB). Impairment of the RNAi pathway in the midgut strongly reduced both midgut barriers for the virus. This confirms that the endogenous RNAi pathway of Ae. aegypti modulates vector competence for SINV in the midgut. The RNAi pathway acts as a gatekeeper to the incoming virus by affecting infection rate of the midgut, intensity of infection, and dissemination from the midgut to secondary tissues.
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影响因子：
4.2
作者：
Campbell CL;Keene KM;Brackney DE;Olson KE;Blair CD;Wilusz J;Foy BD
通讯作者：
Foy BD
影响因子：
2.1
作者：
Cooper, LA;Sina, BJ;Scott, TW
通讯作者：
Scott, TW
影响因子：
3.8
作者：
Adelman, Zach N.;Anderson, Michelle A. E.;Myles, Kevin M.
通讯作者：
Myles, Kevin M.
DOI：
10.1073/pnas.0502815102
发表时间：
2005-09-20
影响因子：
11.1
作者：
Bian, G;Shin, SW;Raikhel, AS
通讯作者：
Raikhel, AS
影响因子：
2.6
作者：
Edwards, MJ;Moskalyk, LA;Jacobs-Lorena, M
通讯作者：
Jacobs-Lorena, M