An assay to image neuronal microtubule dynamics in mice

Tatjana Kleele, Petar Marinković, Philip R. Williams, Sina Stern, Emily E. Weigand, Peter Engerer, Ronald Naumann, Jana Hartmann, Rosa M. Karl, Frank Bradke, Derron Bishop, Jochen Herms, Arthur Konnerth, Martin Kerschensteiner, Leanne Godinho and Thomas Misgeld ()
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Tatjana Kleele: Institute of Neuronal Cell Biology, Technische Universität München, 80802
Petar Marinković: Center for Neuropathology and Prion Research, Ludwig-Maximilians-Universität München, 81377
Philip R. Williams: Institute of Neuronal Cell Biology, Technische Universität München, 80802
Sina Stern: Axon Growth and Regeneration, German Center for Neurodegenerative Diseases (DZNE), 53175
Emily E. Weigand: Indiana University School of Medicine–Muncie
Peter Engerer: Institute of Neuronal Cell Biology, Technische Universität München, 80802
Ronald Naumann: Transgenic Core Facility, Max-Planck-Institute of Molecular Cell Biology and Genetics, 01307
Jana Hartmann: Institute of Neuroscience, Technische Universität München, 80802
Rosa M. Karl: Institute of Neuroscience, Technische Universität München, 80802
Frank Bradke: Axon Growth and Regeneration, German Center for Neurodegenerative Diseases (DZNE), 53175
Derron Bishop: Indiana University School of Medicine–Muncie
Jochen Herms: Center for Neuropathology and Prion Research, Ludwig-Maximilians-Universität München, 81377
Arthur Konnerth: Institute of Neuroscience, Technische Universität München, 80802
Martin Kerschensteiner: Munich Cluster for Systems Neurology (SyNergy), 80336
Leanne Godinho: Institute of Neuronal Cell Biology, Technische Universität München, 80802
Thomas Misgeld: Institute of Neuronal Cell Biology, Technische Universität München, 80802

Nature Communications, 2014, vol. 5, issue 1, 1-10

Abstract: Abstract 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.

Date: 2014
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DOI: 10.1038/ncomms5827

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