Department of Molecular and Cellular Parasitology, Juntendo University, 2-1-1 Hongo, Bunkyo, Tokyo, Japan.
Sci Rep. 2019 Feb 11;9(1):1768. doi: 10.1038/s41598-019-38529-3.
Although it is known that malaria parasites proliferate in the midgut of mosquito vector, their detailed behaviors, from gamete maturation to formation of next generation sporozoite, have not been fully understood at cellular or molecular level. This is mainly attributed to technical difficulties of dissection and whole-mount observation, of delicate and opaque mosquito body contents. In addition, blood pigment surrounding parasites immediately after blood meal also complicates tracing mosquito-stage parasites. Recent revolutionary studies have overcome such negative factors in tissue observation by clearing organisms. CUBIC reagents succeeded to remove both light scattering and blood pigment from various mouse tissues, and to whole-organ image fluorescence-labeled cell structures. In this study, we utilized the advanced version of CUBIC technology and high sensitivity fluorescent markers for see-through observation of mosquito vector after engulfment of rodent malaria parasites to clarify their behaviors during mosquito stage. As a result, we succeeded to visualize oocysts, sporozoites, female gametes and ookinetes in the mosquito bodies without any dissection.
虽然已知疟原虫在蚊子媒介的中肠内增殖，但它们的详细行为，从配子成熟到形成下一代子孢子，在细胞或分子水平上尚未完全了解。这主要归因于解剖和整体观察的技术困难，因为蚊子的身体内容物非常精细且不透明。此外，在进食血液后立即围绕寄生虫的血液色素也使追踪蚊子阶段的寄生虫变得复杂。最近的革命性研究通过透明化方法克服了组织观察中的这些负面因素。CUBIC 试剂成功地从各种小鼠组织中去除了光散射和血液色素，并对荧光标记的细胞结构进行了整体器官成像。在这项研究中，我们利用 CUBIC 技术的高级版本和高灵敏度荧光标记物，对吞噬啮齿动物疟原虫的蚊子媒介进行透明化观察，以阐明它们在蚊子阶段的行为。结果，我们成功地在不进行任何解剖的情况下可视化了蚊子体内的卵囊、子孢子、雌性配子和动合子。
