Molecular and Cellular Bioengineering Research Experiences for Undergraduates (REU) Program
MICROTUBULE-BASED TRANSPORT WITHIN NEURONAL PROCESSES
Efficient, coordinated microtubule-based transport of molecular motor-associated cargoes within axons and dendrites is crucial for neuronal function and survival. We assay neuronal transport function by imaging the movement of various fluorescently-tagged cargoes in cultured cells and in vivo. From these movies, we can generate high-resolution trajectories of labeled cargoes, which allow the extraction of motility parameters such as velocities, pause and reversal frequencies, and cargo densities. By altering specific components of the transport machinery (e.g., motor proteins, cytoskeleton, or cargo components), the cellular environment, or the labeled cargo itself, we will probe the regulation, coordination, and plasticity of the neuromuscular system. Proposed REU projects are designed for a range of skill levels and time commitments, and will provide guided, cross-disciplinary experience in basic cell neurobiology and principles of single-particle and bulk transport. Students will be trained in neuronal cell culture techniques (primary and neuroblastoma cells), and transfection of cultured cells with plasmids suitable for the expression of Fluorescent Protein-fusion proteins. Students will also receive training in wide-field fluorescence microscopy, allowing them to image cargo movement in transfected cells. Students will quantitatively assess transport parameters and neuronal function by analyzing their own movies and/or a growing database of unanalyzed movies using particle-tracking methods. Alternately, advanced students may work on the design of novel fluorescently-tagged proteins using basic molecular cloning techniques. In addition to regular progress meetings with the PI, students will be encouraged to work in teams, particularly on imaging and data analysis. They may also receive broader exposure to related areas of research through periodic journal clubs and lab meetings.
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