Identification of the downstream molecules of agrin/Dok‐7 signaling in muscle

摘要: The development of the neuromuscular junction depends on signaling processes that involve protein phosphorylation. Motor neuron releases agrin to activate muscle Dok-7, a key tyrosine kinase essential for formation mature and functional junction. However, cascade downstream Dok-7 remains poorly understood. In this study, we combined clustered regularly interspaced short palindromic repeats/Cas9 technique quantitative phosphoproteomics analysis study phosphorylation events triggered by agrin/Dok-7. We found level 36 proteins increased specifically stimulation. mutant myotubes, however, 13 failed be enhanced stimulation, suggesting these are Dok-7-dependent tyrosine-phosphorylated proteins, could work as molecules agrin/Dok-7 signaling. validated one Anxa3, in vitro vivo assays. Knocking down Anxa3 cultured myotubes inhibited agrin-induced AChR clustering, whereas reduction mouse muscles induced abnormal postsynaptic development. Collectively, our provides novel insights into complicated network