IRBEH/Spit for Science Publications and Presentations
Document Type
Presentation
Original Publication Date
2025
Date of Submission
June 2025
Abstract
Ninein (Nin) is a microtubule associated protein that has been previously characterized by the Miles Lab for regulating the anxiety-reducing (anxiolytic) effects of alcohol, which is thought to contribute to development of Alcohol Use Disorder (AUD). Previous studies in the lab have investigated the role of Nin deletion on anxiety and ethanol-related behaviors in a conditional knockout model. The following set of studies seek to validate this genetic model. The model was validated using three main methods: immunofluorescence, qRT-PCR, and gel electrophoresis. In PCR studies, Ninein was significantly reduced in cre-expressing animals compared to controls. Gel Electrophoresis confirmed this reduction with the presence of an additional band of a lower molecular weight, indicating the floxed exon (Exon 2) was removed. Additionally, immunofluorescence studies examining Ninein protein expression found subtle changes in Ninein localization in response to cre expression, however further studies are needed to quantify reductions in expression. Ongoing studies are evaluating the use of an alternative translation initiation site using an N-terminal antibody and PCR. Future studies will further investigate alternative Ninein isoforms and their role in ethanol anxiolysis and ethanol consumption.
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Spit for Science Publications and Presentations