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

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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.