Sonoporation Membrane Wounding for High Resolution Imaging of Membrane Repair
The goal of this project is to establish ‘sonoporation’ as a robust and reproducible method for injuring model cells in a way that that facilitates the study of membrane repair. Methods to evaluate the extent of membrane injury and subsequent repair, will be developed in parallel with the sonoporation injury protocol on normal cells, and compared to established methods of membrane wounding. Once sonoporation membrane injury has been established and validated in normal cells, comparisons of the membrane injury and repair capacity of dysferlin deficient cells will be tested. This system is being explicitly designed to be amenable to high resolution microscopic techniques including electron microscopy, which will then be used to follow the membrane repair process in exquisite detail, and should reveal new insights into how and where dysferlin functions in the membrane repair process. By establishing observable “signatures” of the membrane repair process in model cells, we will then be able to look for these same “signatures” in the membrane damage and repair process that occurs in whole skeletal muscle. The knowledge gained from these studies will be used to identify new processes and organelles that influence membrane repair, with the hope of finding new therapeutic targets for dysferlin deficiency.