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利用量子点作为新方法研究伯氏疟原虫在疟疾传播媒介斯氏按蚊中肠的凝集素-碳水化合物识别机制。

Lectin-carbohydrate recognition mechanism of Plasmodium berghei in the midgut of malaria vector Anopheles stephensi using quantum dot as a new approach.

作者信息

Basseri Hamid R, Javazm Mahdi Salari, Farivar Leila, Abai Mohammad R

机构信息

Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, PO Box 6446-14155, Iran.

Department of Medical Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, PO Box 6446-14155, Iran.

出版信息

Acta Trop. 2016 Apr;156:37-42. doi: 10.1016/j.actatropica.2016.01.003. Epub 2016 Jan 6.

DOI:10.1016/j.actatropica.2016.01.003
PMID:26772447
Abstract

Potential targets of Plasmodium ookinetes at the mosquito midgut walls were investigated in relation to interfering malarial transmission. In this study, the essential application of Quantum Dots (QDs) was used to examine the interaction between Plasmodium berghei ookinetes and the Anopheles stephensi midgut, based on lectin-carbohydrate recognition. Two significant lectins were utilized to determine this interaction. Two QDs, cadmium telluride (CdTe)/CdS and cadmium selenide (CdSe)/CdS, were employed in staining Plasmodium ookinete to study its interaction in the midgut of the mosquito vector in vivo. Concurrently, two lectins, wheat germ agglutinin (WGA) and concanavalin A (Con A), were inadvertently exploited to mask lectin binding sites between ookinetes and mosquito midgut cells. The numbers of ookinetes in both lumen and epithelial cells were eventually counted, following adequate preparation of wax sections extracted from whole midgut, and subsequent examination using a differential interference contrast a fluorescence microscopic technique. Interestingly, we detected that neither of the QDs mutated ookinete invasion into the midgut cells of the investigated mosquitoes. QD staining of ookinetes remained permanent despite the effective embedding procedure. The massive binding potency of ookinetes to midgut cells of the cross-examined mosquitoes undoubtedly revealed that Con A did not interrupt ookinete penetration into the midgut wall. In contrast, WGA inhibited ookinete invasion into the midgut cells. The results proved that QD nanoparticles are biocompatible, non-toxic to P. berghei and stable to photobleaching. The QDs staining, which was successfully implemented for ookinete labelling, is a simple and effective tool which plays a crucial role in bioimaging including the study of parasite-vector interactions.

摘要

为了干扰疟疾传播,研究了疟原虫动合子在蚊子中肠壁的潜在靶点。在本研究中,基于凝集素-碳水化合物识别,利用量子点(QD)的重要应用来检测伯氏疟原虫动合子与斯氏按蚊中肠之间的相互作用。使用了两种重要的凝集素来确定这种相互作用。使用碲化镉(CdTe)/硫化镉(CdS)和硒化镉(CdSe)/硫化镉两种量子点对疟原虫动合子进行染色,以研究其在体内蚊子媒介中肠的相互作用。同时,无意中利用了两种凝集素,即麦胚凝集素(WGA)和伴刀豆球蛋白A(Con A)来掩盖动合子与蚊子中肠细胞之间的凝集素结合位点。在对从整个中肠提取的蜡切片进行充分制备,并随后使用微分干涉对比荧光显微镜技术进行检查后,最终对管腔和上皮细胞中的动合子数量进行了计数。有趣的是,我们检测到这两种量子点均未改变动合子侵入所研究蚊子中肠细胞的情况。尽管进行了有效的包埋程序,动合子的量子点染色仍然持久。动合子与交叉检查蚊子中肠细胞的大量结合能力无疑表明,Con A不会干扰动合子穿透中肠壁。相比之下,WGA抑制动合子侵入中肠细胞。结果证明,量子点纳米颗粒具有生物相容性,对伯氏疟原虫无毒且对光漂白稳定。成功用于动合子标记的量子点染色是一种简单有效的工具,在生物成像包括寄生虫-媒介相互作用的研究中起着至关重要的作用。

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