Probing cytoskeletal modulation of passive and active intracellular dynamics using nanobody-functionalized quantum dots

Eugene A. Katrukha, Marina Mikhaylova, Hugo X. van Brakel, Paul M. van Bergen en Henegouwen, Anna Akhmanova, Casper C. Hoogenraad and Lukas C. Kapitein ()
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Eugene A. Katrukha: Cell Biology, Faculty of Science, Utrecht University
Marina Mikhaylova: Cell Biology, Faculty of Science, Utrecht University
Hugo X. van Brakel: Cell Biology, Faculty of Science, Utrecht University
Paul M. van Bergen en Henegouwen: Cell Biology, Faculty of Science, Utrecht University
Anna Akhmanova: Cell Biology, Faculty of Science, Utrecht University
Casper C. Hoogenraad: Cell Biology, Faculty of Science, Utrecht University
Lukas C. Kapitein: Cell Biology, Faculty of Science, Utrecht University

Nature Communications, 2017, vol. 8, issue 1, 1-8

Abstract: Abstract The cytoplasm is a highly complex and heterogeneous medium that is structured by the cytoskeleton. How local transport depends on the heterogeneous organization and dynamics of F-actin and microtubules is poorly understood. Here we use a novel delivery and functionalization strategy to utilize quantum dots (QDs) as probes for active and passive intracellular transport. Rapid imaging of non-functionalized QDs reveals two populations with a 100-fold difference in diffusion constant, with the faster fraction increasing upon actin depolymerization. When nanobody-functionalized QDs are targeted to different kinesin motor proteins, their trajectories do not display strong actin-induced transverse displacements, as suggested previously. Only kinesin-1 displays subtle directional fluctuations, because the subset of microtubules used by this motor undergoes prominent undulations. Using actin-targeting agents reveals that F-actin suppresses most microtubule shape remodelling, rather than promoting it. These results demonstrate how the spatial heterogeneity of the cytoskeleton imposes large variations in non-equilibrium intracellular dynamics.

Date: 2017
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DOI: 10.1038/ncomms14772

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