Resolving bundled microtubules using anti-tubulin nanobodies

Marina Mikhaylova, Bas M. C. Cloin, Kieran Finan, Robert van den Berg, Jalmar Teeuw, Marta M. Kijanka, Mikolaj Sokolowski, Eugene A. Katrukha, Manuel Maidorn, Felipe Opazo, Sandrine Moutel, Marylin Vantard, Frank Perez, Paul M. P. van Bergen en Henegouwen, Casper C. Hoogenraad, Helge Ewers () and Lukas C Kapitein ()
Additional contact information
Marina Mikhaylova: Faculty of Science, Utrecht University, Padualaan 8
Bas M. C. Cloin: Faculty of Science, Utrecht University, Padualaan 8
Kieran Finan: King’s College London, Guy’s Campus
Robert van den Berg: Faculty of Science, Utrecht University, Padualaan 8
Jalmar Teeuw: Faculty of Science, Utrecht University, Padualaan 8
Marta M. Kijanka: Faculty of Science, Utrecht University, Padualaan 8
Mikolaj Sokolowski: Faculty of Science, Utrecht University, Padualaan 8
Eugene A. Katrukha: Faculty of Science, Utrecht University, Padualaan 8
Manuel Maidorn: University of Göttingen, Humboldtallee 23
Felipe Opazo: University of Göttingen, Humboldtallee 23
Sandrine Moutel: Institut Curie, Research Center, 26, rue d'Ulm
Marylin Vantard: Inserm, U836, F-38000, Grenoble, France. Univ. Grenoble Alpes, Grenoble Institut des Neurosciences
Frank Perez: Institut Curie, Research Center, 26, rue d'Ulm
Paul M. P. van Bergen en Henegouwen: Faculty of Science, Utrecht University, Padualaan 8
Casper C. Hoogenraad: Faculty of Science, Utrecht University, Padualaan 8
Helge Ewers: King’s College London, Guy’s Campus
Lukas C Kapitein: Faculty of Science, Utrecht University, Padualaan 8

Nature Communications, 2015, vol. 6, issue 1, 1-7

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms8933

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