Analysing the role of FGF signalling in pluripotency and skeletal muscle development.

在发育过程中，全能干细胞产生一系列特化的分化细胞类型。这一过程涉及发育潜能的逐步限制，并且需要发育胚胎中特定细胞群中的调节基因组高度协调地激活以响应细胞信号。成纤维细胞生长因子（FGF）信号通路是发育过程中基因表达的重要调节因子。我们以前的工作表明，FGF信号激活并维持关键发育调节因子的表达，如转录因子MyoD;生肌调节因子，可以协调骨骼肌细胞从干细胞分化的过程。在脊椎动物的发育过程中，骨骼肌细胞来源于胚胎后部的干细胞库。最近的证据表明，干细胞的特性受到许多多能性因子的调节，包括Oct 4、c-Myc、Lin 28和Nanog。本项目将利用非哺乳类脊椎动物模型（非洲爪蟾和斑马鱼）研究FGF信号、MyoD和多能性因子在体内肌肉发育中的相互作用，并利用青蛙和鱼类胚胎的优势研究发育过程中的细胞谱系、细胞信号和基因表达。我们将在青蛙和鱼类中使用CRISPR/Cas9介导的基因编辑来研究FGF信号传导和多能性因子c-Myc，Lin 28和Vent 2（nanog）在肌肉发育中的作用。通过药理学抑制FGF受体和诱导型FGF受体来过度刺激FGF信号通路，将是揭示发育过程中肌肉决定、维持和分化调节的初步方法。学生也将对青蛙胚胎进行胚胎学操作，以确定后中胚层细胞的多能性。

During development, totipotent stem cells give rise to a host ofspecialised, differentiated cell types. This process involves theprogressive restriction of developmental potential, and requires the highlycoordinated activation of sets of regulatory genes in specific groups ofcells in the developing embryo in response to cell signalling. Thefibroblast growth factor (FGF) signalling pathway is an importantregulator of gene expression during development. Our previous workindicates that FGF signalling activates and maintains the expression ofkey developmental regulators, such as the transcription factor MyoD; amyogenic regulatory factor that can orchestrate the process ofdifferentiation of skeletal muscle cells from stem cells. During vertebratedevelopment skeletal muscle cells arise from a pool of stem cells in theposterior of the developing embryo. Recent evidence indicates thatproperties of stem cells are regulated by a number of pluripotencyfactors, including Oct4, c-Myc, Lin28, and Nanog. This project willinvestigate the interplay of FGF signalling, MyoD and pluripotency factorsin muscle development in vivo using non-mammalian vertebrate animalmodels (Xenopus and zebrafish).This project will take make use of the advantages of frog and fishembryos for investigating cell lineage, cell signalling and gene expressionduring development. We will make use CRISPR/Cas9 mediated geneediting in frogs and fish to investigate the role of the FGF signalling andthe pluripotency factors c-Myc, Lin28, and Vent2 (nanog) in skeletalmuscle development. Manipulating the FGF signalling usingpharmacological inhibition to inactivate and inducible FGF receptors tooverstimulate the pathway will be the initial approach to uncovering theregulation of muscle determination, maintenance and differentiationduring development. The student will also undertake someembryological manipulations of frogs embryos to determine thepluripotent nature of the cells in the posterior mesoderm.