Relative Levels of Drice Expression During Metamorphosis In Larval Fat Bodies of Drosophila melanogaster
Date
2023-07-14
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Abstract
Programmed cell death (PCD) is the self-destruction of a cell as part of development or maintaining homeostasis. The main drivers of apoptosis are a family of enzymes called caspases. The two main caspases in Drosophila are Dronc and Drice. Dronc is an apical caspase, which means it is directly activated by apoptotic signals and in turn activates Drice. Drice is an effector caspase, which means it cleaves certain molecules in the cell which lead to cell death. During Drosophila pupation, larval salivary glands die at 10-12 hours after puparium formation (APF). Unlike most larval tissues, the larval fat bodies of Drosophila do not undergo apoptosis. Instead, they remodel, going from a sheet of fat cells to individual cells at around 12 hours APF. Therefore, it would be expected that larval fat bodies would have much lower levels of Drice than larval salivary glands. In this project, relative levels of Drice protein between Drosophila larval salivary glands and larval fat bodies at 0, 6, 10, and 12 hours APF were measured using Western blotting. It was hypothesized that at all time points levels of Drice protein would be lower in the fat bodies than in the salivary glands. Unfortunately, difficulties with the Western blotting protocol and the antibody used led to a lack of statistically significant results. However, troubleshooting of the experiment did lead to revisions of the Woodard Lab Western blotting protocol that will increase the efficacy and efficiency of future blots. Despite the issues with the project, future study in this area would be very valuable and should make use of the updated Western blotting protocol.
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Keywords
apoptosis, Drosophila melanogaster, Western blot, Drice