Implementation of an In Vivo FRET Sensor for RhoA Activation
Poster Number
50
College
College of Arts and Sciences
Department
Biology
Faculty Mentor
Eric Birgbauer, Ph.D.
Abstract
Axon guidance is an important factor during development, in which nerve cells send out axons to attach to their respective targets. This process is mediated by a plethora of factors and signaling molecules. These factors direct axonal growth through both attraction and repulsion. Furthermore, in greater concentrations, some repulsive factors can cause the growth cone located at the tip of a growing axon to collapse and retract. This phenomenon plays a large role in inhibiting neural injuries from healing. The molecule we are interested in is a bioactive lipid – lysophosphatidic acid (LPA). LPA works through a pathway that results in the activation of RhoA which, in turn, results in cytoskeletal rearrangements that cause growth cone collapse. The detection of molecules like RhoA has been greatly improved through use of techniques with fluorescence resonance energy transfer (FRET). FRET proteins can be used to visualize the presence of various factors because they can be made to change colors, in real time, when a certain molecule is activated. The chief goal of this project is to develop and implement a delivery system for an externally engineered RhoA FRET sensor, then use said sensor to examine RhoA activation in growth cones during growth cone collapse assays. We are using an electroporation-based method that is not yet completed; however, we will perform growth cone collapse experiments in which RhoA activation is observed using the FRET sensor in time-lapse assays within the near future.
Start Date
24-4-2015 3:20 PM
End Date
24-4-2015 4:50 PM
Implementation of an In Vivo FRET Sensor for RhoA Activation
Richardson Ballroom
Axon guidance is an important factor during development, in which nerve cells send out axons to attach to their respective targets. This process is mediated by a plethora of factors and signaling molecules. These factors direct axonal growth through both attraction and repulsion. Furthermore, in greater concentrations, some repulsive factors can cause the growth cone located at the tip of a growing axon to collapse and retract. This phenomenon plays a large role in inhibiting neural injuries from healing. The molecule we are interested in is a bioactive lipid – lysophosphatidic acid (LPA). LPA works through a pathway that results in the activation of RhoA which, in turn, results in cytoskeletal rearrangements that cause growth cone collapse. The detection of molecules like RhoA has been greatly improved through use of techniques with fluorescence resonance energy transfer (FRET). FRET proteins can be used to visualize the presence of various factors because they can be made to change colors, in real time, when a certain molecule is activated. The chief goal of this project is to develop and implement a delivery system for an externally engineered RhoA FRET sensor, then use said sensor to examine RhoA activation in growth cones during growth cone collapse assays. We are using an electroporation-based method that is not yet completed; however, we will perform growth cone collapse experiments in which RhoA activation is observed using the FRET sensor in time-lapse assays within the near future.
