Intestinal expression of H+V-ATPase in the mosquito Aedes albopictus is tightly associated with gregarine infection
Intestinal expression of H+V-ATPase in the mosquito Aedes albopictus is tightly associated with gregarine infection
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DOI：
10.1111/j.1550-7408.2005.00086.x
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发表时间：
2006-03-01
影响因子：
2.2
通讯作者：
Chen, WJ
中科院分区：
文献类型：
--
作者：
Huang, CG;Tsai, KH;Chen, WJ
胞膜ATP酶（V-ATP酶）是一个依赖ATP的质子泵家族，表达于真核细胞的质膜和内膜上。由V-ATP酶介导的细胞内区室（如溶酶体、内体和寄生虫空泡）的酸化对于许多包膜病毒的进入以及细胞内寄生虫侵入或逃离宿主细胞是必不可少的。在蚊子幼虫中，V-ATPase在调节前中肠的碱化中起作用。从白纹伊蚊幼虫组织中提取RNA，克隆了V-ATPase A亚基mRNA的全长序列，并进行了表达。用该融合蛋白制备兔抗V-ATPase多克隆抗体，检测蚊幼虫中肠和马氏管中V-ATPase的活性。在中肠内形成一个含寄生虫的空泡，以响应Ascogregarina taiwanensis的入侵，限制滋养体。使用吖啶橙子染色证明液泡内存在酸化。它的结论是gregarine子孢子被释放的V-ATPase供能的高pH环境中摄入的卵囊。释放的子孢子然后侵入后中肠的上皮细胞并发育。V-ATP酶介导的寄生虫空泡的酸化可能有助于空泡的胞吐作用，从而将滋养体与受感染的上皮细胞隔离开来，以便进一步在细胞外发育。
Vacuolar ATPase (V-ATPase) is a family of ATP-dependent proton pumps expressed on the plasma membrane and endomembranes of eukaryotic cells. Acidification of intracellular compartments, such as lysosomes, endosomes, and parasitophorous vacuoles, mediated by V-ATPase is essential for the entry by many enveloped viruses and invasion into or escape from host cells by intracellular parasites. In mosquito larvae, V-ATPase plays a role in regulating alkalization of the anterior midgut. We extracted RNA from larval tissues of Aedes albopictus, cloned the full-length sequence of mRNA of V-ATPase subunit A, which contains a poly-A tail and 2,971 nucleotides, and expressed the protein. The fusion protein was then used to produce rabbit polyclonal antibodies, which were used as a tool to detect V-ATPase in the midgut and Malpighian tubules of mosquito larvae. A parasitophorous vacuole was formed in the midgut in response to invasion by Ascogregarina taiwanensis, confining the trophozoite(s). Acidification was demonstrated within the vacuole using acridine orange staining. It is concluded that gregarine sporozoites are released by ingested oocysts in the V-ATPase-energized high-pH environment. The released sporozoites then invade and develop in epithelial cells of the posterior midgut. Acidification of the parasitophorous vacuoles may be mediated by V-ATPase and may facilitate exocytosis of the vacuole confining the trophozoites from the infected epithelial cells for further extracellular development.