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.