Structure - Function Relationships in Protozoan Parasites Utilising High Resolution 3D Bioimaging

锥虫科包含许多重要的人类感染性原生动物寄生虫。这些疾病包括在中美洲和南美洲引起恰加斯病的克氏锥虫，据报道，在一些最贫穷的社区有约1800万人感染，没有长期有效的药物治疗，特别是对慢性感染患者。布氏锥虫是一种密切相关的寄生虫，导致非洲昏睡病，每年在撒哈拉以南非洲造成约5万人死亡。布氏锥虫的牛种具有破坏性的经济影响，由于传播疾病的采采蝇的流行，大面积的农村地区无法使用。这两种寄生虫都在世界卫生组织的17种被忽视的热带病名单上，需要更多的努力来了解这些寄生虫的生物学，以帮助找到有效的治疗方法。有两个目标，4个主要研究领域，将在现代寄生虫分子和细胞生物学的背景下发展，并使用新的三维电子显微镜。这些寄生虫在细胞表面称为鞭毛袋，这是分子、营养物质和药物进入的主要场所，并且是这些寄生虫致病性的关键因素。它具有复杂的三维结构，我们的目标是使用一系列三维电子显微镜技术来真正了解鞭毛口袋和相关结构是如何划分和发挥作用的。这些寄生虫已经完成了基因组计划和一系列遗传工具，可以对单个基因进行深入分析。英国和巴西的合作伙伴都是了解这两种寄生虫鞭毛口袋结构的专家。然而，通过双方的共同努力，正在出现重要的分歧。这些差异和相似之处是重要的理解，特别是因为鞭毛口袋是药物和营养物质进入这些细胞的主要场所。我们的目标是运行一个完全合作的项目，以进一步详细了解这些差异，3年来工作人员和学生之间的访问设计旨在最大限度地提高4个实验研究领域的合作伙伴之间的联系和讨论。我们还将在第二年组织一个小型研讨会，以确保实验结果和结论可以传播给广泛的学术受众，包括那些从事药物发现，内吞作用（细胞中的营养吸收）和免疫学的人。为了真正了解活细胞如何分裂并与周围环境相互作用，我们需要了解它们的三维空间组织的详细视图。由于现有的电子显微镜固有的局限性，这很难实现。最近，三维电子显微镜以自动化方式对生物体细胞和组织的详细结构进行成像，并且该提案的合作伙伴是这些3D显微镜技术的领先中心。受过训练的科学家的数量在国内和国际上都很低，在3年的资助期间，我们的目标是培训尽可能多的员工和学生，因为我们相信这些技术将在未来10年内彻底改变普通细胞生物学。

The family Trypanosomatidae contains a number of important human-infective protozoan parasites. These include Trypanosoma cruzi which causes Chagas Disease in Central and South America and there are reported to be ~18 million people infected in some of the poorest communities with no long term effective drug treatment, particularly for chronically infected patients. Trypanosoma brucei is a closely related parasite that causes African sleeping sickness and kills ~50,000 people annually in Sub-Saharan Africa. The cattle species of Trypanosoma brucei has a devastating economic impact, with large areas of countryside that cannot be used due to the prevalence of the Tsetse fly, which spreads the disease. Both parasites are on the World Health Organisation list of 17 neglected Tropical Diseases and more efforts are required to understand the biology of these parasites to help find effective treatments.There are two aim objectives with 4 main research areas that will be developed within a context of modern molecular and cell biology of parasites and the use of novel three dimensional electron microscopy.These parasites have an indentation in the cell surface called the flagellar pocket, which is the major site of entry and exit of molecules, nutrients and entry of drugs and is a critical factor in the pathogenicity of these parasites. It has a complex three dimensional structure and we aim to use a range of three dimensional electron microscopy techniques to really understand how the flagellar pocket and associated structures divides and functions. These parasites have completed genome projects and a range of genetic tools to allow in-depth analysis of individual genes. The partners in both the UK and Brazil are experts in understanding the flagellar pocket structure of the two parasites. However, important differences are emerging through concerted efforts by both partners. These differences and similarities are important to understand, particularly as the flagellar pocket is a major site for entry of drugs and nutrients to these cells. Our objective is to run a fully collaborative project to understand these differences in further detail and the design of the visits between staff and students over the 3 years is aimed at maximising contact and discussion between the partners in the context of 4 experimental research areas. We will also organise a mini-symposium during year 2 to ensure that the outcomes of the experiments and conclusions can be disseminated to a wide academic audience, including those working with drug discovery, endocytosis (nutrient uptake in cells) and immunology.In order to really understand how living cells divide and interact with their surroundings we need to understand a detailed view of their three dimensional spatial organisation. This has been difficult to achieve using existing electron microscopes due to their inherent limitations. Recently, there have been significant developments in three dimensional electron microscopy to image the detailed structure of cells and tissues of organisms in an automated manner and the partners on this proposal are leading centres of excellence for these 3D microscopy techniques. The number of trained scientists is low both nationally and internationally and during the 3 year grant our aim is to train as many staff and students as possible as we believe these techniques will revolutionise general cell biology in the next 10 years.