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11. Nuclear poly-glutamine aggregates rupture the nuclear envelope and hinder its repair

Author

Korsten, Giel, Osinga, Miriam, Pelle, Robin A., Serweta, Albert K., Hoogenberg, Baukje, Kampinga, Harm H., Kapitein, Lukas C., Korsten, Giel, Osinga, Miriam, Pelle, Robin A., Serweta, Albert K., Hoogenberg, Baukje, Kampinga, Harm H., and Kapitein, Lukas C.

Abstract

Huntington's disease (HD) is caused by a polyglutamine expansion of the huntingtin protein, resulting in the formation of polyglutamine aggregates. The mechanisms of toxicity that result in the complex HD pathology remain only partially understood. Here, we show that nuclear polyglutamine aggregates induce nuclear envelope (NE) blebbing and ruptures that are often repaired incompletely. These ruptures coincide with disruptions of the nuclear lamina and lead to lamina scar formation. Expansion microscopy enabled resolving the ultrastructure of nuclear aggregates and revealed polyglutamine fibrils sticking into the cytosol at rupture sites, suggesting a mechanism for incomplete repair. Furthermore, we found that NE repair factors often accumulated near nuclear aggregates, consistent with stalled repair. These findings implicate nuclear polyQ aggregate-induced loss of NE integrity as a potential contributing factor to Huntington's disease and other polyglutamine diseases.

Published
2024

17. Feedback-Driven Assembly of the Axon Initial Segment

Author

Fréal, Amélie, Rai, Dipti, Tas, Roderick P., Pan, Xingxiu, Katrukha, Eugene A., van de Willige, Dieudonnée, Stucchi, Riccardo, Aher, Amol, Yang, Chao, Altelaar, A.F. Maarten, Vocking, Karin, Post, Jan Andries, Harterink, Martin, Kapitein, Lukas C., Akhmanova, Anna, and Hoogenraad, Casper C.

Published
2019
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23. Efficient switching of mCherry fluorescence using chemical caging

Author

Cloin, Bas M. C., De Zitter, Elke, Salas, Desiree, Gielen, Vincent, Folkers, Gert E., Mikhaylova, Marina, Bergeler, Maike, Krajnik, Bartosz, Harvey, Jeremy, Hoogenraad, Casper C., Van Meervelt, Luc, Dedecker, Peter, and Kapitein, Lukas C.

Published
2017

29. Ten-fold Robust Expansion Microscopy

Author

Damstra, Hugo G.J., Mohar, Boaz, Eddison, Mark, Akhmanova, Anna, Kapitein, Lukas C., Tillberg, Paul W., Damstra, Hugo G.J., Mohar, Boaz, Eddison, Mark, Akhmanova, Anna, Kapitein, Lukas C., and Tillberg, Paul W.

Abstract

Expansion microscopy (ExM) is a powerful technique to overcome the diffraction limit of light microscopy that can be applied in both tissues and cells. In ExM, samples are embedded in a swellable polymer gel to physically expand the sample and isotropically increase resolution in x, y, and z. By systematic exploration of the ExM recipe space, we developed a novel ExM method termed Ten-fold Robust Expansion Microscopy (TREx) that, as the original ExM method, requires no specialized equipment or procedures. TREx enables ten-fold expansion of both thick mouse brain tissue sections and cultured human cells, can be handled easily, and enables high-resolution subcellular imaging with a single expansion step. Furthermore, TREx can provide ultrastructural context to subcellular protein localization by combining antibody-stained samples with off-the-shelf small molecule stains for both total protein and membranes.

Published
2023

30. GelMap: intrinsic calibration and deformation mapping for expansion microscopy

Author

Damstra, Hugo G.J., Passmore, Josiah B., Serweta, Albert K., Koutlas, Ioannis, Burute, Mithila, Meye, Frank J., Akhmanova, Anna, Kapitein, Lukas C., Damstra, Hugo G.J., Passmore, Josiah B., Serweta, Albert K., Koutlas, Ioannis, Burute, Mithila, Meye, Frank J., Akhmanova, Anna, and Kapitein, Lukas C.

Abstract

Expansion microscopy (ExM) is a powerful technique to overcome the diffraction limit of light microscopy by physically expanding biological specimen in three dimensions. Nonetheless, using ExM for quantitative or diagnostic applications requires robust quality control methods to precisely determine expansion factors and to map deformations due to anisotropic expansion. Here we present GelMap, a flexible workflow to introduce a fluorescent grid into pre-expanded hydrogels that scales with expansion and reports deformations. We demonstrate that GelMap can be used to precisely determine the local expansion factor and to correct for deformations without the use of cellular reference structures or pre-expansion ground-truth images. Moreover, we show that GelMap aids sample navigation for correlative uses of expansion microscopy. Finally, we show that GelMap is compatible with expansion of tissue and can be readily implemented as a quality control step into existing ExM workflows.

Published
2023

31. A kinesin-based approach for inducing chromosome-specific mis-segregation in human cells

Author

Sub Cell Biology, Celbiologie, Truong, My Anh, Cané-Gasull, Paula, de Vries, Sippe G, Nijenhuis, Wilco, Wardenaar, René, Kapitein, Lukas C, Foijer, Floris, Lens, Susanne, Sub Cell Biology, Celbiologie, Truong, My Anh, Cané-Gasull, Paula, de Vries, Sippe G, Nijenhuis, Wilco, Wardenaar, René, Kapitein, Lukas C, Foijer, Floris, and Lens, Susanne

Published
2023

32. GelMap: intrinsic calibration and deformation mapping for expansion microscopy

Author

Sub Cell Biology, Cell Biology, Neurobiology and Biophysics, Damstra, Hugo G.J., Passmore, Josiah B., Serweta, Albert K., Koutlas, Ioannis, Burute, Mithila, Meye, Frank J., Akhmanova, Anna, Kapitein, Lukas C., Sub Cell Biology, Cell Biology, Neurobiology and Biophysics, Damstra, Hugo G.J., Passmore, Josiah B., Serweta, Albert K., Koutlas, Ioannis, Burute, Mithila, Meye, Frank J., Akhmanova, Anna, and Kapitein, Lukas C.

Published
2023

33. Ten-fold Robust Expansion Microscopy

Author

Sub Cell Biology, Cell Biology, Neurobiology and Biophysics, Damstra, Hugo G.J., Mohar, Boaz, Eddison, Mark, Akhmanova, Anna, Kapitein, Lukas C., Tillberg, Paul W., Sub Cell Biology, Cell Biology, Neurobiology and Biophysics, Damstra, Hugo G.J., Mohar, Boaz, Eddison, Mark, Akhmanova, Anna, Kapitein, Lukas C., and Tillberg, Paul W.

Published
2023

34. GelMap: intrinsic calibration and deformation mapping for expansion microscopy

Author

TN groep Meye, Brain, The Neural Circuits Underlying Stress Eating, Damstra, Hugo G J, Passmore, Josiah B, Serweta, Albert K, Koutlas, Ioannis, Burute, Mithila, Meye, Frank J, Akhmanova, Anna, Kapitein, Lukas C, TN groep Meye, Brain, The Neural Circuits Underlying Stress Eating, Damstra, Hugo G J, Passmore, Josiah B, Serweta, Albert K, Koutlas, Ioannis, Burute, Mithila, Meye, Frank J, Akhmanova, Anna, and Kapitein, Lukas C

Published
2023

35. A kinesin-based approach for inducing chromosome-specific mis-segregation in human cells

Author

CMM, CMM Groep Lens, Cancer, Truong, My Anh, Cané-Gasull, Paula, de Vries, Sippe G., Nijenhuis, Wilco, Wardenaar, René, Kapitein, Lukas C., Foijer, Floris, Lens, Susanne M.A., CMM, CMM Groep Lens, Cancer, Truong, My Anh, Cané-Gasull, Paula, de Vries, Sippe G., Nijenhuis, Wilco, Wardenaar, René, Kapitein, Lukas C., Foijer, Floris, and Lens, Susanne M.A.

Published
2023

43. Direct observation of motor protein stepping in living cells using MINFLUX

Author

Deguchi, Takahiro, primary, Iwanski, Malina K., additional, Schentarra, Eva-Maria, additional, Heidebrecht, Christopher, additional, Schmidt, Lisa, additional, Heck, Jennifer, additional, Weihs, Tobias, additional, Schnorrenberg, Sebastian, additional, Hoess, Philipp, additional, Liu, Sheng, additional, Chevyreva, Veronika, additional, Noh, Kyung-Min, additional, Kapitein, Lukas C., additional, and Ries, Jonas, additional

Published
2023
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45. TRIM46 Controls Neuronal Polarity and Axon Specification by Driving the Formation of Parallel Microtubule Arrays

Author

van Beuningen, Sam F.B., Will, Lena, Harterink, Martin, Chazeau, Anaël, van Battum, Eljo Y., Frias, Cátia P., Franker, Mariella A.M., Katrukha, Eugene A., Stucchi, Riccardo, Vocking, Karin, Antunes, Ana T., Slenders, Lotte, Doulkeridou, Sofia, Sillevis Smitt, Peter, Altelaar, A.F. Maarten, Post, Jan A., Akhmanova, Anna, Pasterkamp, R. Jeroen, Kapitein, Lukas C., de Graaff, Esther, and Hoogenraad, Casper C.

Published
2015
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47. Local changes in microtubule network mobility instruct neuronal polarization and axon specification

Author

Burute, Mithila, Jansen, Klara I, Mihajlovic, Marko, Vermonden, Tina, Kapitein, Lukas C, Celbiologie, Pharmaceutics, Sub Cell Biology, Afd Pharmaceutics, Celbiologie, Pharmaceutics, Sub Cell Biology, and Afd Pharmaceutics

Subjects
Neurons, Cultured, Multidisciplinary, Cells, Hippocampus/metabolism, Kinesins, Cell Polarity, Cell Polarity/physiology, Axons/metabolism, Hippocampus, Microtubules, Axons, Microtubules/metabolism, Neurons/physiology, Cells, Cultured
Abstract

The polarization of neurons into axons and dendrites depends on extracellular cues, intracellular signaling, cytoskeletal rearrangements, and polarized transport, but the interplay between these processes during polarization remains unresolved. Here, we show that axon specification is determined by differences in microtubule network mobility between neurites, regulated by Rho guanosine triphosphatases (GTPases) and extracellular cues. In developing neurons, retrograde microtubule flow prevents the entry of the axon-selective motor protein Kinesin-1 into most neurites. Using inducible assays to control microtubule network flow, we demonstrate that local inhibition of microtubule mobility is sufficient to guide Kinesin-1 into a specific neurite, whereas long-term global inhibition induces the formation of multiple axons. We furthermore show that extracellular mechanical cues and intracellular Rho GTPase signaling control the local differences in microtubule network flow. These results reveal a novel cytoskeletal mechanism for neuronal polarization.

Published
2022

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