It is known that the protein Fondue (Fon) is involved in clotting in Drosophila melanogaster. Consequently, Fon was being studied to... Show moreIt is known that the protein Fondue (Fon) is involved in clotting in Drosophila melanogaster. Consequently, Fon was being studied to characterize its role in the clot. During this study, it was found that Fon and the protein Tiggerin have the same expression pattern. Additionally, mutants for both proteins develop into long, thin pupae (as compared to wild type). Due to this similarity in mutant phenotype and protein localization in combination with literature citing Tiggerin to be involved in both the clot and muscle attachment, Fon was looked at for a muscle attachment phenotype. Furthermore, whilst we do know that Tiggerin is involved in the clot as well as muscle attachment, its specific role in coagulation and its muscle attachment phenotype have not been previously characterized. Due to the pleotropic role of Fon and Tiggerin in muscle attachment and clotting, the long, thin pupal phenotype was used as an indicator of potential clotting mutants in a search at the Bloomington stock center. There, 722 lines were found to have long, thin pupae and from these the 9 lines with the strongest mutant phenotypes were selected to test for coagulation and/or muscle attachment abnormalities. Our collaborator took confocal micrographs of third instar larval fillets obtained from these lines as a means of visualizing actin structure within the muscles. From these images, it was found that the Cchl mutant line had the most severely detached muscles and, consequently, this line was selected as the best candidate for a clotting and muscle attachment mutation and was subjected to further study. The role of a third protein, transglutaminase (TG), which is also involved in clotting, has not been fully characterized due to the lack of a null line. To address this deficiency and fully characterize the role of TG in coagulation in Drosophila, a line with an inactivated TG gene was used as a starting line to generate a null line. M.S. in Biology, May 2014 Show less
