Abstract

Cerebral malaria (CM) is a life-threatening complication of the malaria disease caused by Plasmodium falciparum infection and is responsible for the death of half a million people annually. The molecular pathogenesis underlying CM in humans is not completely understood, although sequestration of infected erythrocytes in cerebral microvessels is thought to play a major role. In contrast, experimental cerebral malaria (ECM) models in mice have been thought to be distinct from human CM, and are mainly caused by inflammatory mediators. Here, to understand the spatial distribution and the potential sequestration of parasites in the whole-brain microvessels during a mouse model of ECM, we utilized the new tissue-clearing method CUBIC (Clear, Unobstructed, Brain/Body Imaging Cocktails and Computational analysis) with light-sheet fluorescent microscopy (LSFM), and reconstructed images in three dimensions (3D). We demonstrated significantly greater accumulation of Plasmodium berghei ANKA (PbANKA) parasites in the olfactory bulb (OB) of mice, compared with the other parts of the brain, including the cerebral cortex, cerebellum and brainstem. Furthermore, we show that PbANKA parasites preferentially accumulate in the brainstem when the OB is surgically removed. This study therefore not only highlights a successful application of CUBIC tissue-clearing technology to visualize the whole brain and its microvessels during ECM, but it also shows CUBIC’s future potential for visualizing pathological events in the whole ECM brain at the cellular level, an achievement that would greatly advance our understanding of human cerebral malaria.

中文翻译：

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使用一种新的三维 CUBIC 组织清除方法检查实验性脑疟疾期间的大脑

摘要

脑型疟疾 (CM) 是由恶性疟原虫引起的疟疾疾病的危及生命的并发症感染，每年造成 50 万人死亡。人类 CM 的分子发病机制尚不完全清楚，尽管脑微血管中受感染的红细胞的隔离被认为起主要作用。相比之下，小鼠实验性脑疟疾 (ECM) 模型被认为与人类 CM 不同，主要由炎症介质引起。在这里，为了了解在 ECM 小鼠模型期间寄生虫在全脑微血管中的空间分布和潜在隔离，我们使用了新的组织清除方法 CUBIC（清晰、通畅、脑/身体成像鸡尾酒和计算分析）与光片荧光显微镜 (LSFM)，以及三维 (3D) 的重建图像。我们展示了显着更大的积累与大脑其他部位（包括大脑皮层、小脑和脑干）相比，小鼠嗅球 (OB) 中的伯氏疟原虫ANKA ( Pb ANKA) 寄生虫。此外，我们表明，当手术切除 OB 时， Pb ANKA 寄生虫优先积聚在脑干中。因此，这项研究不仅突出了 CUBIC 组织清除技术在 ECM 期间可视化整个大脑及其微血管的成功应用，而且还展示了 CUBIC 未来在细胞水平上可视化整个 ECM 大脑中的病理事件的潜力，这一成就将极大地促进我们对人类脑型疟疾的了解。