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Investigating robo1 and robo2 Expression in a Drosophila Injury Model

Declan Barrett, Dr. Hemlata Mistry, Department of Biochemistry and Molecular Biology, Widener University, 1 University Place, Chester PA, 19013

Cellular trauma and the severing of critical neural connections affect diverse biological processes such as the formation and maintenance of axonal trajectories and synaptic connections. To better understand the cellular and molecular mechanisms underlying human spinal cord trauma, we are using the embryonic central nervous system of Drosophila melanogaster. This system is an excellent model for our investigation since there are several established tools to study genetic, cellular, and molecular mechanisms involved in trauma. We used RNA-seq analysis to identify genes whose expression is significantly altered in our injury model, of which robo3 is upregulated. The Robo receptor family contributes to axon guidance, among other developmental processes, and is comprised of the genes robo1, robo2, and robo3. Our objective is to monitor the effects of injury using a monoclonal antibody against Robo (13C9) and determine gene regulation change with respect to robo1 and robo2
 
We severed the ventral nerve cords of late-stage embryos using a fine glass needle. Using the monoclonal antibody against Robo (13C9) we can monitor the axonal proliferation of the CNS. We were able to detect some injured nerve cords, but it is difficult to visualize the extent of the injury epifluorescently. We plan to use confocal microscopy to allow us to better monitor the epitope and see the injury in more detail. We will also use gene expression analysis to determine the effects of injury on the expression of robo1 and robo2. Coupled with this analysis will be a visualization of the Robo proteins in situ. We will vary the times between the injury and analysis to determine if our previous results were not optimal because there was not enough time for protein to be generated and visualized.




Additional Abstract Information

Presenter: Declan Barrett

Institution: Widener University

Type: Poster

Subject: Biology

Status: Approved


Time and Location

Session: Poster 3
Date/Time: Mon 4:30pm-5:30pm
Session Number: 3043