Protein Fingerprinting and Impaired Neurotransmitter Release
This module will allow our students to use protein gel electrophoresis to explore evolution at the molecular level and allow students to study defects in neurotransmitter release. Current laboratory exercises do not address cell structure or physiology. Protein determination, enzyme kinetics, and basic bioinformatics of proteins are used. Allows the student to look at proteins within cells and allow the student to draw correlations between mutant and wildtype proteins and their effect on animal behavior. This multidisciplinary approach is not currently covered in the laboratory. Students will be exposed to protein profiles from selected tissues. Each student will propose a hypothesis, design an experiment based on available samples. The student will be asked to test their hypothesis using SDS-PAGE electrophoresis. Using this technique, the students should be able to compare the band pattern of prominent cellular proteins. Results will help identify similarities, and/or differences, in the protein profile of their chosen samples. In their report, the student will be asked to accept or reject the hypothesis and sustain with arguments (from primary literature) their conclusion. Students will be provided with wild type and several mutant Drosophila fruit fly lines with defects in neurotransmission, specifically, the Shaker potassium channel mutant. This module intends to provide a link between genotype, phenotype and behavior. Mutant and wild type flies will be observed and compared and the severity of the defects will be assessed by the students. The student will then choose Western Blot analysis and/or immunostaining protocols. Students can generate novel results and apply their findings to evolutionary relationships by constructing simple phylogenetic relationship trees from their data. Based on their results, students can decide whether their hypothesis is correct and share their data to complete their analysis through a cooperative learning approach. All students’ results will be posted online on WebCT. They can also compare their results to that in literature. Their laboratory report will be in the format of a research article drawing from their literacy module. A questionnaire on research content, skills, tools and techniques will be offered on the first day of lab and after completing the lab module.