RUI: Membrane Dynamics During Cortical Development in Tetrahymena thermophila

智力优势：生物学中一个更有趣的问题是，看似简单的细胞如何组装和定位复杂的结构，以进行喂养、运动和繁殖。通过靶向膜重塑事件，PI已经确定了影响皮质细胞器模式的基因突变。进一步的分析揭示了一个新的过程，细胞将膜结合的“微泡”（小包的蛋白质和RNA分子）挤压到外部世界。近年来，微囊泡作为一种新的发育和生理反应的信号传导机制而受到广泛关注，并代表了一种调节细胞表面模式生成分子的潜在手段。本项目的工作将利用正常和突变四膜虫细胞中微泡形成的高分辨率显微成像来开发这些结构如何形成，功能和促进皮质细胞器模式的机制细节。微囊泡将进一步纯化用于化学分析，以了解微囊泡的组成。更广泛的影响：圣奥拉夫学院是一所本科院校，致力于通过分享发现过程来教授科学。NSF基金将支持4名本科生的研究活动，特别是在2014年夏季。在生物学方面，学生将接受最先进的荧光和共聚焦显微镜的培训，并获得透射电子显微镜的经验。在化学方面，学生将接受细胞器分离、SDS-PAGE分析和利用系里的质谱仪进行蛋白质组学分析的培训。重点放在第一手的本科生使用的最先进的仪器，为学生的生产科学事业做好准备。沿着这条路线，本科生将进一步受到挑战，在会议和出版物上展示他们的研究结果。总之，这个项目的科学和更广泛的影响将开发一种新的途径，细胞用来交流和响应环境信息，并促进下一代科学家的培训。

Intellectual Merit:One of the more interesting questions in biology is how seemingly simple cells assemble and position complex structures for feeding, movement, and reproduction. The PI has identified genetic mutations that affect the patterning of cortical organelles, through targeting membrane-remodeling events. Further analyses reveal a novel process by which cells extrude membrane-bound "micro-vesicles" (small packets of protein and RNA molecules) into the outside world. Micro-vesicles have recently gained widespread attention in that they represent a new class of signaling mechanism to communicate developmental and physiological responses, and represent a potential means of regulating cell-surface, pattern-generating molecules. The work in this project will utilize high-resolution microscopic imaging of micro-vesicle formation in both normal and mutant Tetrahymena cells to develop mechanistic detail on how these structures form, function, and contribute to the patterning of cortical organelles. Micro-vesicles will further be purified for chemical analyses, to understand vesicle composition. Broader impact: St. Olaf College is an undergraduate institution, devoted to the teaching of science by sharing in the process of discovery. NSF funds will support the research activities of 4 undergraduates, particularly during the summer of 2014. In Biology, students will be trained in state-of-the-art fluorescence and confocal microscopy as well as gain experience with transmission electron microscopy. In Chemistry, students will be trained in organelle isolation, SDS-PAGE analysis, and will utilize the department's mass spectrometer for proteomic analyses. An emphasis is placed on first-hand undergraduate use of state-of-the-art instrumentation, to prepare students for productive scientific careers. Along this line, undergraduates will further be challenged to present their results at meetings and in publications. Together, the scientific and broader impacts of this project will develop a novel pathway that cells use to communicate and respond to environmental information, and foster the training of the next generation of scientists.