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

在骨骼肌发育和修复过程中，肌源性干细胞增殖，当触发这样做时，退出细胞周期并融合形成多核肌纤维。这一过程既发生在胚胎发育过程中，也发生在肌肉损伤或疾病状态（如肌肉萎缩症）时的再生过程中。细胞信号传导是调节肌源性细胞从增殖到分化的重要触发器。该项目将研究MAPK信号通路中一种名为ZAK的蛋白质的作用。该蛋白的基因编码最近与人类遗传性肌病有关，该项目将使用遗传学和分子胚胎学来确定zak在骨骼肌发育和修复中的任何作用。本项目的分析将在体内进行，使用非哺乳动物无脊椎动物模型（爪蟾和斑马鱼）。使用crispr /Cas9进行基因编辑在青蛙和鱼类中非常有效，这种基因靶向方法将用于破坏青蛙和鱼类中的ZAK。利用显性负性和诱导性FGF受体操纵FGF信号来抑制和过度刺激MAPK通路将是解剖需要ZAK活性的通路的一种方法。IGF和EGF信号也将在ZAK调节肌肉发育、生长和修复的背景下进行研究。学生还将获得使用青蛙胚胎的胚胎学操作和在斑马鱼中产生遗传系的经验，以及使用细胞和分子分析信号转导和基因表达的经验。

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.