Cellular signals in the nervous system create a large demand for new proteins at synapses far from the cell body. Neurons interpret these signals and increase protein synthesis (translation) near the synapses in axons and dendrites. This translation localized to axons and dendrites supports proper nervous system development and repair as well as basic brain functions including learning, memory. Importantly, the specific proteins that are synthesized locally in dendrites and axons is specified by mRNA sequences that specify trafficking to these distinct locations. Despite this key relationship of mRNA trafficking to brain function, many fundamental principles of how mRNA trafficking occurs and is regulated are unknown. Studies of mRNA trafficking within the cytoplasm pose unique challenges and require single cell, even single molecule analysis in real time in living cells. This project develops an accessible single molecule assay with a well-documented mRNA as a paradigm that can be applied to training of graduate, undergraduate and high school students. Using these assays the investigators will correlate changes in mRNA movement that occur after interference with cis-acting sequences or trans-acting factors that affect the trafficking of this mRNA in growing neurons. These results will uncover the basis for how mRNA traffics to sites of translation.
The integration of the research with the training of the next generation of scientists is important to the broader impact of this proposal. This project will fundamentally advance our understanding of neuronal cell biology while at the same time train the highest quality scientist/educators for this area of research that is so important to the future scientific efforts of the country.
