Dendrites In Vitro and In Vivo Contain Microtubules of Opposite Polarity and Axon Formation Correlates with Uniform Plus-End-Out Microtubule Orientation.
- Yau KW Cell Biology, Faculty of Science, Utrecht University, 3584 CH, Utrecht, The Netherlands, and.
- Schätzle P Cell Biology, Faculty of Science, Utrecht University, 3584 CH, Utrecht, The Netherlands, and.
- Tortosa E Cell Biology, Faculty of Science, Utrecht University, 3584 CH, Utrecht, The Netherlands, and.
- Pagès S Department of Basic Neurosciences, Faculty of Medicine and the Center for Neuroscience, University of Geneva, 1211 Geneva, Switzerland.
- Holtmaat A Department of Basic Neurosciences, Faculty of Medicine and the Center for Neuroscience, University of Geneva, 1211 Geneva, Switzerland.
- Kapitein LC Cell Biology, Faculty of Science, Utrecht University, 3584 CH, Utrecht, The Netherlands, and c.hoogenraad@uu.nl l.kapitein@uu.nl.
- Hoogenraad CC Cell Biology, Faculty of Science, Utrecht University, 3584 CH, Utrecht, The Netherlands, and c.hoogenraad@uu.nl l.kapitein@uu.nl.
- 2016-01-29
Published in:
- The Journal of neuroscience : the official journal of the Society for Neuroscience. - 2016
cytoskeleton
dendrites
development
microtubule dynamics
neuron
polarity
Animals
Axons
Cell Polarity
Cells, Cultured
Centrioles
Cerebral Cortex
Dendrites
Green Fluorescent Proteins
Hippocampus
Humans
In Vitro Techniques
Mice
Mice, Transgenic
Microfilament Proteins
Microtubule-Associated Proteins
Microtubules
Neurons
RNA, Small Interfering
Rats
Time Factors
Tubulin
English
UNLABELLED
In cultured vertebrate neurons, axons have a uniform arrangement of microtubules with plus-ends distal to the cell body (plus-end-out), whereas dendrites contain mixed polarity orientations with both plus-end-out and minus-end-out oriented microtubules. Rather than non-uniform microtubules, uniparallel minus-end-out microtubules are the signature of dendrites in Drosophila and Caenorhabditis elegans neurons. To determine whether mixed microtubule organization is a conserved feature of vertebrate dendrites, we used live-cell imaging to systematically analyze microtubule plus-end orientations in primary cultures of rat hippocampal and cortical neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neurons in the somatosensory cortex of living mice. In vitro and in vivo, all microtubules had a plus-end-out orientation in axons, whereas microtubules in dendrites had mixed orientations. When dendritic microtubules were severed by laser-based microsurgery, we detected equal numbers of plus- and minus-end-out microtubule orientations throughout the dendritic processes. In dendrites, the minus-end-out microtubules were generally more stable and comparable with plus-end-out microtubules in axons. Interestingly, at early stages of neuronal development in nonpolarized cells, newly formed neurites already contained microtubules of opposite polarity, suggesting that the establishment of uniform plus-end-out microtubules occurs during axon formation. We propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization.
SIGNIFICANCE STATEMENT
Live-cell imaging was used to systematically analyze microtubule organization in primary cultures of rat hippocampal neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neuron in somatosensory cortex of living mice. In vitro and in vivo, all microtubules have a plus-end-out orientation in axons, whereas microtubules in dendrites have mixed orientations. Interestingly, newly formed neurites of nonpolarized neurons already contain mixed microtubules, and the specific organization of uniform plus-end-out microtubules only occurs during axon formation. Based on these findings, the authors propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization.
In cultured vertebrate neurons, axons have a uniform arrangement of microtubules with plus-ends distal to the cell body (plus-end-out), whereas dendrites contain mixed polarity orientations with both plus-end-out and minus-end-out oriented microtubules. Rather than non-uniform microtubules, uniparallel minus-end-out microtubules are the signature of dendrites in Drosophila and Caenorhabditis elegans neurons. To determine whether mixed microtubule organization is a conserved feature of vertebrate dendrites, we used live-cell imaging to systematically analyze microtubule plus-end orientations in primary cultures of rat hippocampal and cortical neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neurons in the somatosensory cortex of living mice. In vitro and in vivo, all microtubules had a plus-end-out orientation in axons, whereas microtubules in dendrites had mixed orientations. When dendritic microtubules were severed by laser-based microsurgery, we detected equal numbers of plus- and minus-end-out microtubule orientations throughout the dendritic processes. In dendrites, the minus-end-out microtubules were generally more stable and comparable with plus-end-out microtubules in axons. Interestingly, at early stages of neuronal development in nonpolarized cells, newly formed neurites already contained microtubules of opposite polarity, suggesting that the establishment of uniform plus-end-out microtubules occurs during axon formation. We propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization.
SIGNIFICANCE STATEMENT
Live-cell imaging was used to systematically analyze microtubule organization in primary cultures of rat hippocampal neurons, dentate granule cells in mouse organotypic slices, and layer 2/3 pyramidal neuron in somatosensory cortex of living mice. In vitro and in vivo, all microtubules have a plus-end-out orientation in axons, whereas microtubules in dendrites have mixed orientations. Interestingly, newly formed neurites of nonpolarized neurons already contain mixed microtubules, and the specific organization of uniform plus-end-out microtubules only occurs during axon formation. Based on these findings, the authors propose a model in which the selective formation of uniform plus-end-out microtubules in the axon is a critical process underlying neuronal polarization.
- Language
-
- English
- Open access status
- bronze
- Identifiers
-
- DOI 10.1523/JNEUROSCI.2430-15.2016
- PMID 26818498
- Persistent URL
- https://sonar.ch/global/documents/220640
Statistics
Document views: 24
File downloads:
- fulltext.pdf: 0