August 03, 2018
The end of July brought a close to Summer Fellows research and there was no shortage of trail blazing Neuroscience exploration. Connor Loomis’ research is described below:
Mapping the interaction between the synaptic adhesion protein Slitrk2 and PSD-95 via a yeast-two hybrid assay
Mentor: Jennifer Round
In the developing nervous system, it is crucial that neurons synapse with their correct partners
to establish proper brain wiring. When synapses form incorrectly, neurological diseases such as schizophrenia and bipolar disorder can occur. Synaptic adhesion proteins are present at synapses to help the presynaptic axon connect with the correct postsynaptic target. Our laboratory studies a specific transmembrane synaptic adhesion protein called Slitrk2. Previously, we discovered that Slitrk2 binds with the MAGUK (membrane-associated guanylate kinase) scaffolding protein, PSD-95, which helps anchor postsynaptic membrane proteins. To determine which domains of Slitrk2 and PSD-95 are essential for the interaction, we performed a yeast-two hybrid assay. Specifically, we constructed DNA plasmids coding for full-length and truncated versions of Slitrk2 and PSD-95. We then transformed these plasmids into yeast to allow the cells to transcribe and translate the proteins. Attached to these proteins are transcription factor domains that when successfully brought together initiate the transcription of reporter genes, allowing the yeast to grow on selective media. Growth in the presence of a yeast toxin and LacZ expression indicate a positive interaction. From this assay, we established that the last 4 amino acids of Slitrk2 are required for interaction with PSD-95. Currently, we are testing which domain of PSD-95 binds to Slitrk2. Future experiments will examine the functional significance of the Slitrk2-PSD-95 interaction on Slitrk2 trafficking and synapse formation.
This program allowed students to pursue independent scholarly projects under the tutelage of a faculty mentor over an eight week period. The faculty mentor helped shape and direct the research and weekly common hour meetings were scheduled. A presentation of the research was held on July 27th, the last day of the program. See more Neuroscience research on the Summer Fellows page of our website. https://www.ursinus.edu/academics/neuroscience/independent-learning/summer-fellows/