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Optogenetic control of organelle transport and positioning
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Journal Article
Optogenetic control of organelle transport and positioning
2015
Overview
An optogenetic strategy allowing light-mediated recruitment of distinct cytoskeletal motor proteins to specific organelles is established; this technique enabled rapid and reversible activation or inhibition of the transport of organelles such as peroxisomes, recycling endosomes and mitochondria with high spatiotemporal accuracy, and the approach was also applied to primary neurons to demonstrate optical control of axonal growth by recycling endosome repositioning.
Light-touch manipulation of cellular organelles
How does the position of organelles within a cell influence cellular functions? In the absence of strategies to control intracellular organelle positioning with spatiotemporal precision, it has been difficult to answer this question. Lukas Kapitein and colleagues have developed an optogenetic strategy, based on light-mediated recruitment of distinct cytoskeletal motor proteins to their specific cargo organelles that allows such cellular manipulations. Using the new technique it is possible to rapidly and reversibly activate or inhibit the transport of specific organelles and demonstrate local modulation of organelle distributions — including peroxisomes, recycling endosomes and mitochondria — with high spatiotemporal accuracy. The authors demonstrate local modulation of organelle distributions for peroxisomes, recycling endosomes and mitochondria. They also applied this approach in primary neurons to establish optical control of axon outgrowth.
Proper positioning of organelles by cytoskeleton-based motor proteins underlies cellular events such as signalling, polarization and growth
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. 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.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 14/35
/ 42/109
/ 42/35
/ 45/22
/ 45/77
/ 82/80
/ 96/63
/ Accuracy
/ Animals
/ Biological Transport - radiation effects
/ Cell Compartmentation - physiology
/ Cell Compartmentation - radiation effects
/ Cytoskeleton - radiation effects
/ Dendritic Spines - metabolism
/ Dendritic Spines - radiation effects
/ Dynein
/ Endosomes - radiation effects
/ Humanities and Social Sciences
/ Intracellular Space - metabolism
/ Intracellular Space - radiation effects
/ Kinesin
/ Kinesins - radiation effects
/ letter
/ Light
/ Microtubules - radiation effects
/ Mitochondria - radiation effects
/ Molecular motors (Biochemistry)
/ Motility
/ Myosin
/ Myosin Type V - radiation effects
/ Neurons
/ Peroxisomes - radiation effects
/ Proteins
/ Rats
/ Science
