title: An assay to image neuronal microtubule dynamics in mice.
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.
authors.affiliation: Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Kleele
authors.affiliation: 1] Center for Neuropathology and Prion Research, Ludwig-Maximilians-Universität München, 81377 Munich, Germany [2] German Center for Neurodegenerative Diseases (DZNE), 80336 Munich, Germany.
authors.forename: Petar
authors.initials: P
authors.lastname: Marinković
authors.affiliation: Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany.
authors.forename: Philip R
authors.initials: PR
authors.lastname: Williams
authors.affiliation: Axon Growth and Regeneration, German Center for Neurodegenerative Diseases (DZNE), 53175 Bonn, Germany.
authors.forename: Sina
authors.initials: S
authors.lastname: Stern
authors.affiliation: Department of Physiology, Indiana University School of Medicine-Muncie, Muncie, Indiana 47306, USA.
authors.forename: Emily E
authors.initials: EE
authors.lastname: Weigand
authors.affiliation: Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Engerer
authors.affiliation: Transgenic Core Facility, Max-Planck-Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
authors.forename: Ronald
authors.initials: R
authors.lastname: Naumann
authors.affiliation: Institute of Neuroscience, Technische Universität München, 80802 Munich, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Hartmann
authors.affiliation: Institute of Neuroscience, Technische Universität München, 80802 Munich, Germany.
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Karl
authors.affiliation: Axon Growth and Regeneration, German Center for Neurodegenerative Diseases (DZNE), 53175 Bonn, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Bradke
authors.affiliation: Department of Physiology, Indiana University School of Medicine-Muncie, Muncie, Indiana 47306, USA.
authors.forename: Derron
authors.initials: D
authors.lastname: Bishop
authors.affiliation: 1] Center for Neuropathology and Prion Research, Ludwig-Maximilians-Universität München, 81377 Munich, Germany [2] German Center for Neurodegenerative Diseases (DZNE), 80336 Munich, Germany [3] Munich Cluster for Systems Neurology (SyNergy), 80336 Munich, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Herms
authors.affiliation: 1] Institute of Neuroscience, Technische Universität München, 80802 Munich, Germany [2] Munich Cluster for Systems Neurology (SyNergy), 80336 Munich, Germany [3] Center of Integrated Protein Science (CIPSM), 81377 Munich, Germany.
authors.forename: Arthur
authors.initials: A
authors.lastname: Konnerth
authors.affiliation: 1] Munich Cluster for Systems Neurology (SyNergy), 80336 Munich, Germany [2] Institute of Clinical Neuroimmunology, Ludwig-Maximilians Universität München, 81377 Munich, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Kerschensteiner
authors.affiliation: Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany.
authors.forename: Leanne
authors.initials: L
authors.lastname: Godinho
authors.affiliation: 1] Institute of Neuronal Cell Biology, Technische Universität München, 80802 Munich, Germany [2] German Center for Neurodegenerative Diseases (DZNE), 80336 Munich, Germany [3] Munich Cluster for Systems Neurology (SyNergy), 80336 Munich, Germany [4] Center of Integrated Protein Science (CIPSM), 81377 Munich, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Misgeld
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001681:Biological Assay; D016764:Cell Polarity; D004195:Disease Models, Animal; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure*; D057054:Molecular Imaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D061251:Primary Cell Culture; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018800:Thy-1 Antigens / Q000235:genetics / Q000378:metabolism; D014741:Video Recording
chemical_list: D001426:Bacterial Proteins; C522423:EB3 protein, mouse; D008164:Luminescent Proteins; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D018800:Thy-1 Antigens; C054562:yellow fluorescent protein, Bacteria