A cross-species study of the effects of agrochemicals on microtubule dynamics through the development of new live imaging probes

通过开发新型活体成像探针进行农用化学品对微管动力学影响的跨物种研究

• 批准号: 1621711
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1621711
• 关键词: cross; species; study; effects; agrochemicals

Global demand for food is growing; current projections indicate that we will need 60% more food in 2050. This puts increasing pressure on a food production system that already has to cope with reduced natural resources and the results of climate change. To help us meet the growing demand for food, it is vital that we increase the efficiency with which we grow crops. An important method to increase crop productivity is the careful use of agrochemicals. Indeed, the use of such agrochemicals has provided significant advances in crop production and food provision. It is therefore essential that we continue to develop better and safer agrochemicals that will help us make the best use of our limited natural resources.The microtubule cytoskeleton is an important biological target for the development of new herbicides and fungicides. Microtubules are required for a wide range of cellular processes, from cell division to cellular transport, making them an important target for agrochemical compounds. However, because microtubules are so well conserved across species, a major challenge is to identify compounds that are toxic to the targeted pest species (plants or fungi) without affecting crops, humans and other non-target species. It is therefore vital that we have a better understanding of the differences in the responses to microtubule perturbators in all of these populations. To this end, we need to develop tools that can allow a more detailed assessment of microtubule function across a range of organisms. This information will contribute to early decision-making in the development and design of new agrochemicals. Microtubules are inherently dynamic structures: being constantly remodeled through fluctuations in polymerization and depolymerisation. This dynamic behavior is crucial for many of the cellular functions of microtubules; for example, during cell division the microtubules of the mitotic spindle search for chromosomes by probing the cytoplasm through rapid cycles of polymerization and depolymerization. The suppression of these dynamics is also key for other microtubule functions in the cell; indeed the stabilization of microtubules by specific microtubule associated proteins (MAPs) is vital for determining the polarity of a cell. Microtubule-targeting agrochemicals generally function by disrupting the fine balance of microtubule dynamics in a cell. Because only a subtle change in dynamics can have a profound effect on the cell, it is important that we understand in detail how a compound affects the polymerization, depolymerization and stabilization of microtubules and compare this across both target and non-target organisms. To do this, it is crucial that we are able to visualize and measure microtubule dynamics in living cells, tissues and embryos, rather than dead, fixed tissues.Our project aims to determine in detail the effect of specific agrochemicals on microtubule dynamics in plant and animal cells at the level of the single cell, tissue and whole embryo. Importantly, this will involve the development of new probes for the imaging of microtubules in living cells. These new probes will overcome current limitations with existing probes, which give can give diffuse or uneven staining of microtubules or affect microtubule dynamics. Understanding the biological differences of microtubule perturbers across species at both the level of the cell and whole organism will provide important insights into potential adverse effects of these compounds. This will ultimately aid decision-making and the progression of compounds through the Research and Development pipeline and consequently impact on both the speed to market and the likely success of a compound. 4. Compare the MT responses found in (3) with those seen in the target species of the selectedagrochemicals.

全球对粮食的需求正在增长;目前的预测表明，到2050年，我们对粮食的需求将增加60%。这给已经不得不科普自然资源减少和气候变化后果的粮食生产系统带来了越来越大的压力。为了帮助我们满足日益增长的粮食需求，提高农作物种植效率至关重要。提高作物产量的一个重要方法是谨慎使用农用化学品。事实上，使用这些农用化学品在作物生产和粮食供应方面取得了重大进展。因此，我们必须继续开发更好、更安全的农用化学品，以帮助我们充分利用有限的自然资源。微管细胞骨架是开发新除草剂和杀菌剂的重要生物靶点。微管是从细胞分裂到细胞运输的广泛细胞过程所必需的，使其成为农用化学品化合物的重要靶标。然而，由于微管在物种间保存得如此之好，一个主要的挑战是鉴定对目标害虫物种（植物或真菌）有毒而不影响作物、人类和其他非目标物种的化合物。因此，至关重要的是，我们有一个更好的理解在所有这些人群中的微管扰动的反应的差异。为此，我们需要开发能够更详细地评估一系列生物体中微管功能的工具。这一信息将有助于在开发和设计新的农用化学品的早期决策。微管是固有的动态结构：通过聚合和解聚的波动不断重塑。这种动态行为对于微管的许多细胞功能至关重要;例如，在细胞分裂期间，有丝分裂纺锤体的微管通过聚合和解聚的快速循环探测细胞质来寻找染色体。这些动力学的抑制也是细胞中其他微管功能的关键;事实上，特定微管相关蛋白（MAP）对微管的稳定性对于确定细胞的极性至关重要。微管靶向农用化学品通常通过破坏细胞中微管动力学的精细平衡来起作用。因为只有动力学的细微变化才能对细胞产生深远的影响，所以我们必须详细了解化合物如何影响微管的聚合、解聚和稳定，并在靶生物和非靶生物中进行比较。为此，我们必须能够观察和测量活细胞、组织和胚胎中的微管动力学，而不是死的、固定的组织。我们的项目旨在详细确定特定农用化学品对植物和动物细胞中微管动力学的影响，在单个细胞、组织和整个胚胎的水平上。重要的是，这将涉及开发用于活细胞中微管成像的新探针。这些新的探针将克服现有探针的局限性，现有探针可能会对微管进行扩散或不均匀的染色或影响微管动力学。了解不同物种在细胞和整个生物体水平上微管干扰物的生物学差异将为这些化合物的潜在不良影响提供重要的见解。这最终将有助于决策和化合物通过研发管道的进展，从而影响上市速度和化合物的可能成功。4.将（3）中发现的MT反应与所选化学品的靶物种中观察到的MT反应进行比较。