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

During skeletal muscle development and repair, myogenic stem cells proliferate
and, when triggered to do so, exit the cell cycle and fuse to form
multinucleated myofibres. This process occurs both during embryonic
development and during regenerative processes that happen when muscle is
injured or in a disease-state, as occurs in muscular dystrophy. Cell signalling is
an important trigger that regulates a myogenic cell's switch from proliferation
to differentiation. This project will investigate the role of a protein that is part
of the MAPK signalling pathway, called ZAK. The gene coding for this protein
has recently been implicated in an inherited myopathy in humans and this
project will use genetics and molecular embryology to determine any role for
ZAK in skeletal muscle development and repair.
The analyses in this project will be done in vivo using non-mammalian
vertebrate animal models (Xenopus and zebrafish). Gene editing using
CRISPR/Cas9 is very effective in frogs and fish and this gene targeting approach
will be used to disrupt ZAK in frogs and fish. Manipulating the FGF signalling
with dominant negative and inducible FGF receptors to inhibit and
overstimulate the MAPK pathway will be one approach to dissect the pathways
that require ZAK activity. IGF and EGF signalling will also be investigated in the
context of ZAK regulation of myogenic development , growth and repair. The
student will also gain experience using embryological manipulations of frogs
embryos and generating genetic lines in zebrafish, as well as using cell and
molecular assays to analyse signal transduction and gene expression.