Investigation of the molecular mechanisms of malarial ookinete invasion into the mosquite midgut epithelium in the rodent mararial parasite, Plasmodium berghei

疟原虫动动分子侵入啮齿类疟原虫伯氏疟原虫中肠上皮细胞的分子机制研究

• 批准号: 12470060
• 负责人: YUDA Masao
• 金额: $ 9.15万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470060/
• 关键词: Malaria; mosquite; ookinete; sporozoite; infection; molecular mechanisms; ネズミマラリア; CTRP; ノックアウト原虫; ハマダラカ; 接着侵入

To search for genes playing roles in malarial infection of the mosquito vector, we constructed EST databases of the vector stages of rodent malarial parasite. We selected some genes from these databases and prepared knock-out parasites of each gene. As criteria for selection, we adopted presence of the N-terminal signal peptide, because this indicates that the gene products may play a role in parasite interactions with the vector mosquito. In this study, we identified some genes that play a critical role in malarial parasite infection of mosquito organs. One of them encodes 21kDa protein with a secretory protein-lilke structure and is necessay for ookinete invasion into the midgut epithelium. We performed immuno-electeon microscopy and found that this protein is localized in the micronemes. Other one encodes an attachment protein-like molecule and is expressed by the sporozoite in the oocyst but not by the salivary gland sporozoite. By disruption of this gene, we demonstrated that it is essential for the malarial parasite to infect the mosquito salivary gland.

为了寻找在蚊媒疟疾感染中起作用的基因，我们构建了啮齿类疟原虫媒介阶段的EST数据库。我们从这些数据库中选择了一些基因，并制备了每个基因的敲除寄生虫。作为选择的标准，我们采用了N-末端信号肽的存在，因为这表明基因产物可能在寄生虫与载体蚊子的相互作用中起作用。在这项研究中，我们确定了一些在疟疾寄生虫感染蚊子器官中起关键作用的基因。其中一个编码21 kDa的蛋白质，具有类似分泌蛋白的结构，是动合子侵入中肠上皮所必需的。我们进行了免疫电子显微镜，发现这种蛋白质是本地化的微线。另一个编码附着蛋白样分子，由卵囊中的子孢子表达，但不由唾液腺子孢子表达。通过破坏该基因，我们证明了疟疾寄生虫感染蚊子唾液腺是必需的。

项目成果

Toru Kariu, Masao Yuda, Kazuhiko Yano, Yasuo Chinzei: "MAEBL is essential for malarial sporozoite infection of the mosquito salivary gland"J. Exp. Med.. 195. 1317-1323 (2002)

Toru Kariu、Masao Yuda、Kazuhiko Yano、Yasuo Chinzei：“MAEBL 对于蚊子唾液腺的疟疾子孢子感染至关重要”J.

Haruhiko Isawa, Masao Yuda, Yuki Orito, Yasuo Chinzei: "Mosquito salivary protein inhibits activation of a contact system by bindingto factor XII and high molecular weight kininogen"J. Biol. Chem.. 277. 27651-27658 (2002)

Haruhiko Isawa、Masao Yuda、Yuki Orito、Yasuo Chinzei：“蚊子唾液蛋白通过与因子 XII 和高分子量激肽原结合来抑制接触系统的激活”J。

S.Iwanaga, M.Okada, H.Isawa, A.Morita, M.Yuda, Y.Chinzei: "Identification and characterization of novel salivary thrombin inhibitors from the ixodidae tick, Haemaphysalis longicornis"Eur. J. Biochem.. (in press). (2003)

S.Iwanaga、M.Okada、H.Isawa、A.Morita、M.Yuda、Y.Chinzei：“来自 ixodidae 蜱、Haemaphysalis longicornis 的新型唾液凝血酶抑制剂的鉴定和表征”Eur。

Toru Kariu, Masao Yuda, Kazuhiko Yano and Yasuo Chinzei: "MAEBL is essential for malarial sporozoite infection of the mosquite salivary gland"J.Exp.Med.. 195. 1317-1323 (2002)

Toru Kariu、Masao Yuda、Kazuhiko Yano 和 Yasuo Chinzei：“MAEBL 对于蚊子唾液腺的疟疾子孢子感染至关重要”J.Exp.Med.. 195. 1317-1323 (2002)

Masao Yuda, Kazuhiko Yano, Takafumi Tsuboi, Motomi Torii and Yasuo Chinzei: "von Willebrand Factor A Domain-related Protein : a novel microneme protein of the malaria ookinete highly conserved throughout Plasmodium parasites"Mol.Bolchem.Parasitol.. 116. 6

Masao Yuda、Kazuhiko Yano、Takafumi Tsuboi、Motomi Torii 和 Yasuo Chinzei：“von Willebrand 因子 A 结构域相关蛋白：一种在疟原虫寄生虫中高度保守的疟疾动合子的新型微线体蛋白”Mol.Bolchem.Parasitol.. 116. 6