DMD‐10 and its Potential Effect on Regulation of the Glutamate Receptor GLR‐1 in C. elegans

An organism's nervous system processes decisions on how to respond to their environment. Studying how proteins of the nervous system are regulated enables us to identify how an organism responds to its environment and alters its behavior. In our lab, we study glutamate receptors, specifically GLR‐1. Glutamate is used for communication between neurons, and changes in glutamate receptors are believed to play an important role in learning and memory. The DAF‐7/TGF‐β signaling pathway regulates GLR‐1 in C. elegans and mutants in the DAF‐7/TGF‐β signaling pathway have increased GLR‐1. However, the mechanism through which GLR‐1 levels are increased in the daf‐7 mutant worms is unknown. My research addressed how GLR‐1 levels were increased in the daf‐7 worms. I explored the possibility that the transcription factor DMD‐10 is affected in the daf‐7 mutants, and then in turn affects GLR‐1. We tested this by measuring spontaneous reversals (a GLR‐1 dependent behavior) in both single mutant and double mutant worms. If DMD‐10 is affected in the daf‐7 mutants, we predict that the dmd‐10; daf‐7 double mutants would have fewer reversals than the daf‐7 mutant. The dmd‐10; daf‐7 double mutants did not have fewer spontaneous reversals than the single daf‐7 mutant, which suggests that the DAF‐7/TGF‐β signaling pathway does not use DMD‐10 to alter levels of GLR‐1. However, the dmd‐10 single mutant had fewer reversals than the wildtype, suggesting that DMD‐10 is required for normal reversals. The results of this experiment suggested that although DMD‐10 is not involved in DAF‐7/TGF‐β regulation of GLR‐1, it may have an independent effect on GLR‐1. My current research is aimed at determining if DMD‐10 is involved in the regulation of GLR‐1. I have generated a strain of dmd‐10 mutant worms that expresses GFP‐tagged GLR‐1 (GLR‐1::GFP). Imaging of GLR‐1::GFP in wild type and dmd‐10 mutant worms will allow us to test if DMD‐10 is involved in GLR‐1 regulation. This strain is being analyzed for total levels of GLR‐1::GFP using western blotting and will indicate if DMD‐10 is reducing total levels of GLR‐1. Synaptic levels of GLR‐1::GFP will be analyzed using fluorescence imaging that provides information on the location of GLR‐1 and whether it is localized in the cell body or synapses. Together, these results will indicate if DMD‐10 is affecting GLR‐1 and will give us insight into how regulation is occurring. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .