3D motion of vesicles along microtubules helps them to circumvent obstacles in cells

Ione Verdeny-Vilanova; Fabian Wehnekamp; Nitin Mohan; Ángel Sandoval Álvarez; Joseph Steven Borbely; Jason John Otterstrom; Don C Lamb; Melike Lakadamyali

doi:10.1242/jcs.201178

3D motion of vesicles along microtubules helps them to circumvent obstacles in cells

J Cell Sci. 2017 Jun 1;130(11):1904-1916. doi: 10.1242/jcs.201178. Epub 2017 Apr 18.

Authors

Ione Verdeny-Vilanova¹, Fabian Wehnekamp², Nitin Mohan¹, Ángel Sandoval Álvarez¹, Joseph Steven Borbely¹, Jason John Otterstrom¹, Don C Lamb², Melike Lakadamyali³

Affiliations

¹ ICFO-Institut de Ciències Fotòniques, The Barcelona Institute of Science and Technology, Castelldefels, Barcelona 08860, Spain.
² Ludwig-Maximilians-Universität München, Department of Chemistry, Physical Chemistry, Center for Integrated Protein Science Munich, and Nanosystems Initiative Munich, Butenandtstr. 5-13, München 81377, Germany.
³ ICFO-Institut de Ciències Fotòniques, The Barcelona Institute of Science and Technology, Castelldefels, Barcelona 08860, Spain melikel@mail.med.upenn.edu.

Abstract

Vesicle transport is regulated at multiple levels, including regulation by scaffolding proteins and the cytoskeleton. This tight regulation is essential, since slowing or stoppage of transport can cause accumulation of obstacles and has been linked to diseases. Understanding the mechanisms by which transport is regulated as well as how motor proteins overcome obstacles can give important clues as to how these mechanisms break down in disease states. Here, we describe that the cytoskeleton architecture impacts transport in a vesicle-size-dependent manner, leading to pausing of vesicles larger than the separation of the microtubules. We further develop methods capable of following 3D transport processes in living cells. Using these methods, we show that vesicles move using two different modes along the microtubule. Off-axis motion, which leads to repositioning of the vesicle in 3D along the microtubule, correlates with the presence of steric obstacles and may help in circumventing them.

Keywords: 3D tracking; Motor protein; Super-resolution microscopy; Vesicle trafficking.

MeSH terms

Animals
Cell Line
Chlorocebus aethiops
Cytoplasmic Vesicles / drug effects
Cytoplasmic Vesicles / metabolism*
Cytoplasmic Vesicles / ultrastructure
Cytoskeleton / drug effects
Cytoskeleton / metabolism*
Cytoskeleton / ultrastructure
Epithelial Cells / drug effects
Epithelial Cells / metabolism*
Epithelial Cells / ultrastructure
Gene Expression
Genes, Reporter
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Lysosomal-Associated Membrane Protein 2 / genetics
Lysosomal-Associated Membrane Protein 2 / metabolism*
Lysosomes / drug effects
Lysosomes / metabolism
Lysosomes / ultrastructure
Microspheres
Microtubules / drug effects
Microtubules / metabolism*
Microtubules / ultrastructure
Nocodazole / pharmacology
Optical Imaging
Paclitaxel / pharmacology
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Red Fluorescent Protein
Tubulin / genetics
Tubulin / metabolism*

Substances

LAMP2 protein, human
Luminescent Proteins
Lysosomal-Associated Membrane Protein 2
Recombinant Fusion Proteins
Tubulin
Green Fluorescent Proteins
Paclitaxel
Nocodazole