Resolving bundled microtubules using anti-tubulin nanobodies
2015 | journal article. A publication with affiliation to the University of Göttingen.
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Resolving bundled microtubules using anti-tubulin nanobodies
Mikhaylova, M.; Cloin, B. M. C.; Finan, K.; van den Berg, R.; Teeuw, J.; Kijanka, M. M. & Sokolowski, M. et al. (2015)
Nature Communications, 6 art. 7933. DOI: https://doi.org/10.1038/ncomms8933
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Details
- Authors
- Mikhaylova, Marina; Cloin, Bas M. C.; Finan, Kieran; van den Berg, Robert; Teeuw, Jalmar; Kijanka, Marta M.; Sokolowski, Mikolaj; Katrukha, Eugene A.; Maidorn, Manuel; Opazo, Felipe; Moutel, Sandrine; Vantard, Marylin; Perez, Frank; Henegouwen, Paul M. P. van Bergen En; Hoogenraad, Casper C.; Ewers, Helge; Kapitein, Lukas C.
- Abstract
- Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell-and neurobiology.
- Issue Date
- 2015
- Status
- published
- Publisher
- Nature Publishing Group
- Journal
- Nature Communications
- ISSN
- 2041-1723