title: Optogenetic control of organelle transport and positioning.
abstract: Proper positioning of organelles by cytoskeleton-based motor proteins underlies cellular events such as signalling, polarization and growth. For many organelles, however, the precise connection between position and function has remained unclear, because strategies to control intracellular organelle positioning with spatiotemporal precision are lacking. Here we establish optical control of intracellular transport by using light-sensitive heterodimerization to recruit specific cytoskeletal motor proteins (kinesin, dynein or myosin) to selected cargoes. We demonstrate that the motility of peroxisomes, recycling endosomes and mitochondria can be locally and repeatedly induced or stopped, allowing rapid organelle repositioning. We applied this approach in primary rat hippocampal neurons to test how local positioning of recycling endosomes contributes to axon outgrowth and found that dynein-driven removal of endosomes from axonal growth cones reversibly suppressed axon growth, whereas kinesin-driven endosome enrichment enhanced growth. Our strategy for optogenetic control of organelle positioning will be widely applicable to explore site-specific organelle functions in different model systems.
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Petra
authors.initials: P
authors.lastname: van Bergeijk
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Max
authors.initials: M
authors.lastname: Adrian
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Casper C
authors.initials: CC
authors.lastname: Hoogenraad
authors.affiliation: Cell Biology, Department of Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, the Netherlands.
authors.forename: Lukas C
authors.initials: LC
authors.lastname: Kapitein
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology / Q000528:radiation effects; D001692:Biological Transport / Q000528:radiation effects; D002451:Cell Compartmentation / Q000502:physiology* / Q000528:radiation effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism / Q000528:radiation effects; D049229:Dendritic Spines / Q000378:metabolism / Q000528:radiation effects; D004398:Dyneins / Q000378:metabolism / Q000528:radiation effects; D011992:Endosomes / Q000378:metabolism* / Q000528:radiation effects; D006624:Hippocampus / Q000166:cytology; D042541:Intracellular Space / Q000378:metabolism / Q000528:radiation effects; D016547:Kinesins / Q000378:metabolism / Q000528:radiation effects; D008870:Microtubules / Q000378:metabolism / Q000528:radiation effects; D008928:Mitochondria / Q000378:metabolism* / Q000528:radiation effects; D024701:Myosin Type V / Q000378:metabolism / Q000528:radiation effects; D062308:Optogenetics / Q000379:methods*; D020675:Peroxisomes / Q000378:metabolism* / Q000528:radiation effects; D051381:Rats
chemical_list: D024701:Myosin Type V; D004398:Dyneins; D016547:Kinesins