Role of srGAP3 protein stability in mediating voluntary cocaine abuse in Gridlock'd mutant mice

Elaina Cote

Document Type

Article

Publication Date

Summer 2022

Keywords

JMG

JAX Location

In: Student Reports, Summer 2022, The Jackson Laboratory

Abstract

Slit Robo GTPase activating protein 3 (srGAP3), also known as mental disorder associated GAP, is a cytoskeletal-related protein that regulates actin polymerization in neurons. It is known to play roles in intellectual disability, and may contribute to addiction behaviors. Previous research has shown reduced levels of this protein in mice with the S642P Gridlock’d mutation, as well as altered addiction phenotypes. We hypothesized that the Gridlock’d mutation causes a structural disruption in the srGAP3 protein that causes rapid degradation of the protein.. We used in vitro methods to perform a stability assay which measures the relative rate of srGAP3 degradation in the mutant compared to wild type variant of the protein. Human embryonic kidney (HEK293T) cells were transfected with the srGAP3 gene and harvested at different time points. Western blot was used to detect relative levels of the protein over time to assess half life. Preliminary data does not show srGAP3 degradation over time, however the experimental approach requires further troubleshooting before we can obtain conclusive data. We will continue this project by optimizing the transfection process in hopes of receiving stronger signal from srGAP3 in Western blots.

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