Date of Award

2013

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Keywords

Ethanol, Biochemistry, Cerebellum

Area of Concentration

Biochemistry

Abstract

Alcoholism is an addiction to ethanol (EtOH) that is marked by cyclic periods of chronic EtOH intake, withdrawal, attempts at abstinence, and relapse. Research into the excitotoxic properties of EtOH in the central nervous system (CNS) dominates the current scientific field, while studies are less focused on the mechanisms underlying brain atrophy and damage following chronic EtOH exposure and withdrawal periods. The current thesis attempted to explore the mechanisms following EtOH exposure and withdrawal in an in vitro model using adult (ten- and eleven-week old) 250 μm mouse cerebellar slices. Myelin and axon-associated proteins and actin fibers were significantly degraded following EtOH exposure, consistent with previous literature. However, EtOH exposure significantly decreased the primary metabolic enzyme GAPDH, as well as the water transporter protein AQ4. EtOH-W (4 h EtOH exposure, 20 h control serum) had differential and unexpected effects only on five proteins: degrading three (NFL, Bcl-2, Calpain-1), while increasing two (β-actin, CNPase). Control samples underwent significant protein degradation in the model compared to naive samples that were not exposed to the culture medium, and combined with propidium iodide staining measurements, suggests that a level of basal damage occurred in the plated samples. Model exposure did not completely destroy cerebellar slices, as indicated by PCR products of 4 genes (β-actin, Calpain-1, Calpain-2, Calpastatin). Future studies on in vitro models of adult myelin and axon in chronic alcoholism are needed to elucidate the role of proteases and apoptosis in degradation, but should also account for initial protease degradation.

Rights

The author has granted New College of Florida the nonexclusive right to archive, make accessible, and distribute for educational purposes this work in whole or in part in all forms of media, now or hereafter known. The copyright of this work remains with the author.

Download

Share

COinS