Evaluating TDP-43 Targets in Amyotrophic Lateral Sclerosis Using Drosophila and Patient Spinal Cords
posterposted on 20.04.2020, 23:22 by Alexander Blythe, Erik Lehmkuhl, Suvithanandhini Loganathan, Eric Alsop, Dianne Barrameda, Bhavani B. Siddegowda, Archi Joardar, Kendall Jensen, Daniela C. Zarnescu
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.