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About the Project
To understand how mutations in the thin filament regulatory protein troponin cause familial hypertrophic cardiomyopathy (HCM) it is important to know the structure of the cardiac thin filament. Techniques for the genetic manipulation of the zebrafish are well established and it has become a major model for the study of human cardiac disease. Our lab isolates zebrafish thin filaments from adult fish and are working to create a zebrafish model with mutations which cause HCM in humans. We study the 3D structure using Cryo-Electron Microscopy (EM), where the thin filaments are preserved in a frozen hydrated state. Using transmission EM and single particle analysis we can reconstruct a 3D map of the filament.
We now want to study the structure of these filaments in situ and our goal for this project is to look at frozen hydrated cardiomyocytes from zebrafish. After freezing the myocytes, we will perform cryo-sectioning and tomography of the to elucidate the ultrastructure. Some of the techniques that will be learnt in this project include; microdissection, EM and Cryo-EM sample preparation, EM and Cryo-EM imaging, image processing and 3D molecular fitting.
Funding Notes
Please apply to the Faculty of Life Sciences, School of Physiology and Pharmacology, selecting the programme 'MSc by Research'
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References
Relaxed and active thin filament structures; a new structural basis for the regulatory mechanism.
Paul DM, Squire JM, Morris EP. J Struct Biol. 2017 Mar;197(3):365-371.
Becker JR, Deo R C, Werdlich AA, Panakova D, Coy S and MacRae C A: Human Cardiomyopathy mutations induce myocyte hyperplasia and activate hypertrophic pathways during cardiogenisis in zebrafish. Disease Models & Mechanisms. 2011 4, 400-410
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