Kleele Tatjana, Marinković Petar, Williams Philip R, Stern Sina, Weigand Emily E, Engerer Peter, Naumann Ronald, Hartmann Jana, Karl Rosa M, Bradke Frank, Bishop Derron, Herms Jochen, Konnerth Arthur, Kerschensteiner Martin, Godinho Leanne, Misgeld Thomas
Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany.
1] Center for Neuropathology and Prion Research, Ludwig-Maximilians-Universität München, 81377 Munich, Germany [2] German Center for Neurodegenerative Diseases (DZNE), 80336 Munich, Germany.
Nat Commun. 2014 Sep 12;5:4827. doi: 10.1038/ncomms5827.
Microtubule dynamics in neurons play critical roles in physiology, injury and disease and determine microtubule orientation, the cell biological correlate of neurite polarization. Several microtubule binding proteins, including end-binding protein 3 (EB3), specifically bind to the growing plus tip of microtubules. In the past, fluorescently tagged end-binding proteins have revealed microtubule dynamics in vitro and in non-mammalian model organisms. Here, we devise an imaging assay based on transgenic mice expressing yellow fluorescent protein-tagged EB3 to study microtubules in intact mammalian neurites. Our approach allows measurement of microtubule dynamics in vivo and ex vivo in peripheral nervous system and central nervous system neurites under physiological conditions and after exposure to microtubule-modifying drugs. We find an increase in dynamic microtubules after injury and in neurodegenerative disease states, before axons show morphological indications of degeneration or regrowth. Thus increased microtubule dynamics might serve as a general indicator of neurite remodelling in health and disease.
神经元中的微管动力学在生理、损伤和疾病中起着关键作用，并决定微管的方向，而微管方向是神经突极化的细胞生物学相关因素。几种微管结合蛋白，包括末端结合蛋白3（EB3），特异性地结合到微管不断生长的正端。过去，荧光标记的末端结合蛋白已揭示了体外和非哺乳动物模型生物中的微管动力学。在此，我们设计了一种基于表达黄色荧光蛋白标记的EB3的转基因小鼠的成像检测方法，以研究完整哺乳动物神经突中的微管。我们的方法能够在生理条件下以及在暴露于微管修饰药物后，测量外周神经系统和中枢神经系统神经突中体内和体外的微管动力学。我们发现在损伤后和神经退行性疾病状态下，在轴突显示出退化或再生的形态学迹象之前，动态微管会增加。因此，增加的微管动力学可能是健康和疾病中神经突重塑的一个普遍指标。
