Evaluating TDP-43 Targets in Amyotrophic Lateral Sclerosis Using Drosophila and Patient Spinal Cords
posterposted on 20.04.2020 by Alexander Blythe, Erik Lehmkuhl, Suvithanandhini Loganathan, Eric Alsop, Dianne Barrameda, Bhavani B. Siddegowda, Archi Joardar, Kendall Jensen, Daniela C. Zarnescu
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This poster describes several experiments concerning a potential target of pathological TAR DNA-binding protein 43 (TDP-43) that was singled out as a likely candidate of translational repression following previous tagged ribosomal affinity purification (TRAP) and RNA immunoprecipitation (RIP) experiments. The specific target is dally-like protein, a glypican, which had low association with ribosomes and high levels of co-precipitation with TDP-43. I used a Drosophila model of ALS using a Gal4-UAS system to overexpress TDP-43 in motor neurons, which results in the recapitulation of ALS phenotypes (reduced lifespan, locomotor dysfunction, sleep fragmentation, formation of cytoplasmic complexes in motor neurons, nuclear loss of TDP-43, etc.). I performed a lifespan assay and performed Western blots on nervous tissue (specifically ventral nerve cords and neuromuscular junctions) and used immunohistochemistry to obtain images of DLP at the cellular level. In addition, I performed Western blots on samples of spinal cord tissue from patients whose genomes demonstrated mutations in the TDP-43 gene that are implicated in familial ALS. These samples were by the VA of southern Arizona.