RUI: Spatiotemporal mapping of the membrane trafficking networks involved in secretion and autophagy in the unicellular zygnematophyte, Penium margaritaceum

这项研究项目的目的是提供一个全面的检查细胞内的膜运输网络参与了广泛的细胞外基质(ECM)的加工，该植物是一种单细胞合生植物，或藻类，其祖先产生了陆地植物。这一点很重要，因为目前还没有关于Penium或任何其他合生植物的膜运输网络的详细地图。负责ECM生产的膜贩运网络对古代陆地化(从海上到陆地的移动)至关重要。今天，这些网络负责支持主要淡水群落中的合生植物，包括短暂的水华和生物膜。该项目将通过使用高分辨率的电子显微镜分析来构建细胞内膜的结构模型，并将使用目标化学和抗体试剂来解释特定膜组件的功能。该项目不仅将提供对植物细胞外基质分泌动态和膜运输活动的关键洞察，这些活动是植物向陆地移动的关键，而且还将扩大斯基德莫尔学院本科生和当地高中生利用新开放的整合科学中心(CIS)的新空间和现有设备从事有意义的细胞生物学研究项目的机会。它还将包括一个独特的博士后研究人员培训和指导计划，该计划将参与研究并为在本科学院的潜在职业做准备。该项目将调查与生产广泛的细胞外基质有关的亚细胞机械的结构和功能。Zygnematophycean是一组绿藻，与陆地植物关系最密切，是其祖先。该项目将产生合成膜运输网络的结构和功能的模型，该网络涉及单细胞合生植物珍珠霉的细胞外基质分泌、胞质分裂、空泡动力学和自噬。它将使用多种显微镜工具、断层扫描、免疫标记和实验分析来获取数据，以构建详细的贩运网络地图和3-D模型。这些将为许多合生植物在形成短暂的花朵和生物膜时所表现出的快速细胞生长和相关的高分泌活动提供关键的洞察。它将阐明暴露在各种非生物应激源和靶向化学制剂下的细胞中这些网络的调制。该项目还将对细胞质分裂过程中使用的独特的沟槽-细胞板系统以及植物膜质/细胞板的进化提供重要的见解。最后，它将提供关于基于液泡和自噬的隔室及其在合生植物中的功能作用的新信息。这项研究由生物科学理事会分子和细胞生物科学部门的细胞动力学和功能集群资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The goal of this research project is to provide a comprehensive examination of the membrane trafficking networks inside the cell involved in the processing of the extensive extracellular matrix (ECM) of Penium margaritaceum, a single-celled zygnematophyte, or algae, whose ancestors gave rise to land plants. This is important because at present, no detailed maps of the membrane trafficking network of Penium or any other zygnematophytes exist. Membrane trafficking networks responsible for the production of the ECM were critical for ancient terrestrialization (movement from sea to land). Today, these networks are responsible for supporting zygnematophytes in major freshwater communities, including ephemeral blooms and biofilms. This project will construct structural models of the membranes inside the cell by using high resolution, electron microscopy analyses, and the functions of specific membrane components will be explained using targeted chemical and antibody agents. This project will not only provide critical insight into the ECM secretion dynamics in plants and membrane trafficking events that were key in plant movement to land, but will also expand opportunities for Skidmore College undergraduates and local high school students to engage in a meaningful cell biology research program using new space and available equipment at the newly opened Center for Integrative Sciences (CIS). It will also include a unique training and mentoring program for a postdoctoral researcher that will involve participation in the research and preparation for a potential career at an undergraduate institution.This project will investigate the structure and function of the subcellular machinery associated with the production of the extensive extracellular matrix of Zygnematophycean algae, the group of green algae most closely related and ancestral to land plants. The project will yield models that synthesize the architecture and function of the membrane trafficking networks involved with ECM secretion, cytokinesis, vacuolar dynamics and autophagy in the model unicellular zygnematophyte, Penium margaritaceum. It will use multiple microscopy tools, tomography, immunolabeling and experimental analyses to acquire data for the construction of detailed maps and 3-D models of the trafficking networks. These will provide key insight into the rapid cell growth and associated hyper-secretory activity that many zygnematophytes exhibit when forming ephemeral blooms and biofilms. It will elucidate modulations of these networks in cells exposed to various abiotic stressors and targeted chemical agents. This project will also provide significant insight into the unique furrow-cell plate system employed during cytokinesis and the evolution of the plant phragmoplast/cell plate. Finally, it will provide new information about vacuole- and autophagy-based compartments and their functional roles in zygnematophytes.This research is funded by the Cellular Dynamics and Function cluster in the Division of Molecular and Cellular Biosciences in the Directorate of Biological Sciences.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.