Dissecting a role for ZAK in skeletal muscle development using genetics and embryology in zebrafish and Xenopus

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During skeletal muscle development and repair, myogenic stem cells proliferateand, when triggered to do so, exit the cell cycle and fuse to formmultinucleated myofibres. This process occurs both during embryonicdevelopment and during regenerative processes that happen when muscle isinjured or in a disease-state, as occurs in muscular dystrophy. Cell signalling isan important trigger that regulates a myogenic cell's switch from proliferationto differentiation. This project will investigate the role of a protein that is partof the MAPK signalling pathway, called ZAK. The gene coding for this proteinhas recently been implicated in an inherited myopathy in humans and thisproject will use genetics and molecular embryology to determine any role forZAK in skeletal muscle development and repair.The analyses in this project will be done in vivo using non-mammalianvertebrate animal models (Xenopus and zebrafish). Gene editing usingCRISPR/Cas9 is very effective in frogs and fish and this gene targeting approachwill be used to disrupt ZAK in frogs and fish. Manipulating the FGF signallingwith dominant negative and inducible FGF receptors to inhibit andoverstimulate the MAPK pathway will be one approach to dissect the pathwaysthat require ZAK activity. IGF and EGF signalling will also be investigated in thecontext of ZAK regulation of myogenic development , growth and repair. Thestudent will also gain experience using embryological manipulations of frogsembryos and generating genetic lines in zebrafish, as well as using cell andmolecular assays to analyse signal transduction and gene expression.