Defense Date

2013

Document Type

Thesis

Degree Name

Master of Science

Department

Human Genetics

First Advisor

Mike Grotewiel

Abstract

The genetic pathways influencing alcohol abuse and dependence are poorly characterized. Many critical discoveries about the interactions between ethanol-related behaviors and genetics have been made in the fruit fly Drosophila melanogaster. Coupling the statistical power of model organism studies to human association studies bolsters the analytical efficacy of these genomic approaches. A variety of behavioral assays are available for assessing behavioral responses to ethanol in Drosophila. However, we find our previously described eRING assay is influenced by the commonly used transgenic marker mini-white. We developed a Simple Sedation Assay (SSA) that is insensitive to the effects of white and mini-white. In SSAs, expression of endogenous wild-type white was not necessary for normal responses to ethanol. Neither expression nor RNAi-mediated knockdown of the transgenic mini-white influenced the effects of ethanol in flies. Critically, mini-white expression did not affect the phenotypes of flies with known alterations in ethanol sensitivity. Also, loss of function mutations in Clic show decreased sensitivity to ethanol in both eRING assays (as previously reported) and SSAs. Therefore, we explored the role of the known Clic interactors, TGF-β and ryanodine receptors. These studies were inconclusive but do not exclude the need for future work. Finally, using bioinformatic tools we constructed a mutli-species network of genes predicted to interact with Clic. Our RNAi screen against the Clic network serves as an important proof-of-concept and holds great potential for uncovering important therapeutic targets for alcohol use disorders.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

July 2013

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