OSIB:RUI: Elucidating the cell biology and developmental regulation of sporogenesis and spore dimorphism in the microsporidia Nosema ceranae using a novel flow cytometry approach

一些感染人类和其他对人类活动很重要的动物（如蜜蜂）的微生物病原体很难研究，因为它们的生命周期很复杂，其中许多不同生命阶段的细胞发生在宿主的细胞内。在生物学的其他领域，一种称为流式细胞术的技术已被用于鉴别和分离基于差异染色的复杂混合物中的不同类型的细胞。本研究表明，流式细胞术可用于分离和量化感染和危害蜜蜂及其蜂群的特定寄生虫的不同生命阶段。流式细胞术将用于更好地了解影响寄生虫不同生命阶段产生的宿主和寄生虫因素，以及每个生命阶段如何影响宿主的健康和宿主之间的感染传播。此外，本科研究人员将在流式细胞术和其他技术方面得到支持和培训，为这些学生支持美国生物经济的职业做好准备。具有复杂生命周期的专性细胞内病原体的研究是困难的，因为它们不容易在宿主外繁殖，并且很难分离。这些障碍使许多重要的科学探索途径无法实现，特别是在宿主和病原体都是非模式生物的情况下，可获得的物种特异性分子工具有限或没有，导致科学进展缓慢。流式细胞术结合特殊的细胞染料被发展为一种策略，以促进对蜜蜂的关键病原体微孔虫的了解。假设该策略将分离和量化两种不同发育时间、不同形态属性和不同感染特性的孢子类型对应的蜜蜂生命阶段。孢子类型及其前体的分离将使科学家能够定义细胞身份的分子结构，这有助于它们的独特特性。这些孢子类型和它们在蜜蜂感染期间的前体的量化将允许科学通过定义控制这两种孢子类型产生的环境因素来回答有关感染动力学的关键问题。这项技术的使用有望促进微孢子虫研究的一些新方向。此外，这项工作是确定潜在治疗方法的基础，以防止或减少小孢子虫感染的蜜蜂，一个重要的传粉媒介。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Some microbial pathogens infecting humans and other animals important to human activities, such as honey bees are hard to study because of their complex life cycles, in which cells of many different life stages occur inside the cells of their hosts. In other areas of biology, a technology called flow cytometry has been used to identify and separate cells of different types from a complex mixture based on differential staining. This proposal shows evidence that flow cytometry can be used to isolate and quantify different life stages of a specific parasite that infects and harms honey bees and their colonies. Flow cytometry will be used to better understand host and parasite factors that influence the production of different parasite life stages and how each life stage impacts on the health of the host and spread of infection between hosts. In addition, undergraduate researchers will be supported and trained in flow cytometry and other technologies to prepare these students for careers that support the US bioeconomy.The study of obligate intracellular pathogens with complex life cycles is difficult because they cannot easily be reproduced outside the host and are challenging to isolate. These obstacles render many important avenues of scientific inquiry unachievable, especially in situations where both host and pathogen are non-model organisms for which limited or no species-specific molecular tools are available, leading to slowed scientific progress. Flow cytometry in conjunction with specialized cell dyes was developed as a strategy to advance the understanding of Nosema ceranae, a key pathogen of honey bees. The hypothesis is that this strategy will isolate and quantify N. ceranae life stages corresponding to two spore types with different developmental timing, distinct morphological attributes, and divergent infectious properties. Isolation of the spore types and their precursors will allow scientists to define the molecular architecture of cellular identity that contributes to their unique properties. Quantification of these spore types and their precursors during infection in bees will allow science to answer key questions about infection dynamics by defining environmental factors that govern the generation of the two spore types. Use of this technique promises to facilitate a number of new directions in microsporidia research. In addition, this work is the basis for identifying potential therapeutics to prevent or minimize Nosema infection of bees, an important pollinator.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.