2021-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/675693足體(podosome)與癌細胞的侵襲性偽足(invadopodium)是一種肌動蛋白組成的結構，主要功能為細胞的附著、侵襲轉移以及基質的降解因此與人類的健康與疾病息息相關。然而，我們對足體的動態調控以及它在組織發育上的生理功能並不完全了解。因此，我們將透過研究dynamin-2在足體形成的分子功能以及在肌肉細胞中對神經肌肉接合處發育的影響來回答這個問題。我們實驗室最近發現dynamin-2會獨特地集中在足體的核心周圍，並存在於果蠅及老鼠神經肌肉接合處的突觸後細胞膜。並且，抑制dynamin-2的表達或是其酵素活性，都會造成足體結構的改變。為了進一步了解dynamin-2這個細胞膜重塑蛋白如何利用與肌動蛋白結合以及水解三磷酸鳥苷(GTP)的活性來調控足體的形成及它在肌肉細胞中的功能，我們將利用多學科的實驗方法，包含體外重組、細胞培養以及動物實驗，結合最先進的顯微技術來探討dynamin-2在肌肉細胞中對足體的結構與功能的影響。這個研究計畫的成果將展現dynamin-2在突觸後細胞膜上的新穎功能以及它對足體的形成與更替調控，最後我們將發展出一個可穿透細胞膜的足體抑制胜肽。 Podosome and its cancer counterpart invadopodium are actin-based structures functioning on cell adhesion, invasion and extracellular matrix remodeling thus are critical for our health and diseases. However, the regulation of podosome turnover and its function in tissue development are largely unclear. In this proposal, we plan to investigate the molecular role of a GTPase, dynamin-2, in regulating podosome maturation, turnover and neuromuscular junction (NMJ) morphogenesis. From our preliminary results, we have observed a unique spatial distribution of dynamin-2 in muscular podosome and mouse neuromuscular junction. In addition, the structure of podosome is significantly disrupted while dynamin-2 activity is inhibited. To further understand how dynamin-2 regulates the dynamics of podosome through its actin bundling and GTP hydrolysis activities and how dynamin-2 participates in NMJ maturation, we will utilize multidisciplinary approaches and state-of-the-art microscopy technologies to study the podosome structure and function with in vitro, in cell and in vivo model systems. The results of this proposal will be the first finding which illustrates the function of dynamin-2 in postsynaptic membrane, its molecular role in podosome turnover and develops a cell-penetrable podosome inhibiting peptide.足體更替侵襲性偽足肌肉發生神經肌肉接合處的形態發生肌動蛋白重塑podosome turnoverinvadopodiummuscle developmentneuromuscular junction morphogenesisF-actin remodeling