Interplay between kinesin-1 and cortical dynein during axonal outgrowth and microtubule organization in Drosophila neurons

eLife - Tập 4
Urko del Castillo1, Michael Winding1, Wen Lü1, Vladimir I. Gelfand1
1Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, United States

Tóm tắt

In this study, we investigated how microtubule motors organize microtubules in Drosophila neurons. We showed that, during the initial stages of axon outgrowth, microtubules display mixed polarity and minus-end-out microtubules push the tip of the axon, consistent with kinesin-1 driving outgrowth by sliding antiparallel microtubules. At later stages, the microtubule orientation in the axon switches from mixed to uniform polarity with plus-end-out. Dynein knockdown prevents this rearrangement and results in microtubules of mixed orientation in axons and accumulation of microtubule minus-ends at axon tips. Microtubule reorganization requires recruitment of dynein to the actin cortex, as actin depolymerization phenocopies dynein depletion, and direct recruitment of dynein to the membrane bypasses the actin requirement. Our results show that cortical dynein slides ‘minus-end-out’ microtubules from the axon, generating uniform microtubule arrays. We speculate that differences in microtubule orientation between axons and dendrites could be dictated by differential activity of cortical dynein.

Từ khóa


Tài liệu tham khảo

Ahmad, 1995, Microtubules released from the neuronal centrosome are transported into the axon, Journal of Cell Science, 108 (Pt 8), 2761, 10.1242/jcs.108.8.2761

Ahmad, 1998, Cytoplasmic dynein and dynactin are required for the transport of microtubules into the axon, The Journal of Cell Biology, 140, 391, 10.1083/jcb.140.2.391

Akhmanova, 2015, Microtubule minus-end-targeting proteins, Current Biology, 25, R162, 10.1016/j.cub.2014.12.027

Arthur, 2015, Dendrite arborization requires the dynein cofactor NudE, Journal of Cell Science, 128, 2191, 10.1242/jcs.170316

Baas, 1988, Polarity orientation of microtubules in hippocampal neurons: uniformity in the axon and nonuniformity in the dendrite, Proceedings of the National Academy of Sciences of the United States of America, 85, 8335, 10.1073/pnas.85.21.8335

Baas, 1999, Microtubules and neuronal polarity, Neuron, 22, 23, 10.1016/S0896-6273(00)80675-3

Barlan, 2013, The microtubule-binding protein ensconsin is an essential cofactor of kinesin-1, Current Biology, 23, 317, 10.1016/j.cub.2013.01.008

Clackson, 1998, Redesigning an FKBP-ligand interface to generate chemical dimerizers with novel specificity, Proceedings of the National Academy of Sciences of the United States of America, 95, 10437, 10.1073/pnas.95.18.10437

del Castillo, 2015, Pavarotti/MKLP1 regulates microtubule sliding and neurite outgrowth in drosophila neurons, Current Biology, 25, 200, 10.1016/j.cub.2014.11.008

Egger, 2013, In vitro imaging of primary neural cell culture from drosophila, Nature Protocols, 8, 958, 10.1038/nprot.2013.052

Fink, 2006, Dynein-dependent motility of microtubules and nucleation sites supports polarization of the tubulin array in the fungus ustilago maydis, Molecular Biology of the Cell, 17, 3242, 10.1091/mbc.E05-12-1118

Grabham, 2007, Cytoplasmic dynein and LIS1 are required for microtubule advance during growth cone remodeling and fast axonal outgrowth, Journal of Neuroscience, 27, 5823, 10.1523/JNEUROSCI.1135-07.2007

Hackney, 2000, Kinesin's IAK tail domain inhibits initial microtubule-stimulated ADP release, Nature Cell Biology, 2, 257, 10.1038/35010525

Heim, 2004, Genetically encoded indicators of cellular calcium dynamics based on troponin c and green fluorescent protein, Journal of Biological Chemistry, 279, 14280, 10.1074/jbc.M312751200

Hendershott, 2014, Regulation of microtubule minus-end dynamics by CAMSAPs and patronin, Proceedings of the National Academy of Sciences of the United States of America, 111, 5860, 10.1073/pnas.1404133111

Hoogenraad, 2003, Bicaudal d induces selective dynein-mediated microtubule minus end-directed transport, The EMBO Journal, 22, 6004, 10.1093/emboj/cdg592

Jaqaman, 2008, Robust single-particle tracking in live-cell time-lapse sequences, Nature Methods, 5, 695, 10.1038/nmeth.1237

Jiang, 2014, Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition, Developmental Cell, 28, 295, 10.1016/j.devcel.2014.01.001

Jolly, 2010, Kinesin-1 heavy chain mediates microtubule sliding to drive changes in cell shape, Proceedings of the National Academy of Sciences of the United States of America, 107, 12151, 10.1073/pnas.1004736107

Kapitein, 2015, Building the neuronal microtubule cytoskeleton, Neuron, 87, 492, 10.1016/j.neuron.2015.05.046

Kapitein, 2010, Probing intracellular motor protein activity using an inducible cargo trafficking assay, Biophysical Journal, 99, 2143, 10.1016/j.bpj.2010.07.055

Kim, 2007, Microtubule binding by dynactin is required for microtubule organization but not cargo transport, The Journal of Cell Biology, 176, 641, 10.1083/jcb.200608128

Kiyomitsu, 2013, Cortical dynein and asymmetric membrane elongation coordinately position the spindle in anaphase, Cell, 154, 391, 10.1016/j.cell.2013.06.010

Kotak, 2012, Cortical dynein is critical for proper spindle positioning in human cells, The Journal of Cell Biology, 199, 97, 10.1083/jcb.201203166

Kuznetsov, 1988, The quaternary structure of bovine brain kinesin, The EMBO Journal, 7, 353, 10.1002/j.1460-2075.1988.tb02820.x

Laan, 2012, Cortical dynein controls microtubule dynamics to generate pulling forces that position microtubule asters, Cell, 148, 502, 10.1016/j.cell.2012.01.007

Lee, 2002, Short stop provides an essential link between f-actin and microtubules during axon extension, Development, 129, 1195, 10.1242/dev.129.5.1195

Lin, 2012, Mitotic motors coregulate microtubule patterns in axons and dendrites, Journal of Neuroscience, 32, 14033, 10.1523/JNEUROSCI.3070-12.2012

Liu, 2010, Kinesin-12, a mitotic microtubule-associated motor protein, impacts axonal growth, navigation, and branching, Journal of Neuroscience, 30, 14896, 10.1523/JNEUROSCI.3739-10.2010

Lu, 2013, Organelle transport in cultured Drosophila cells: S2 cell line and primary neurons, Journal of Visualized Experiments, 10.3791/50838

Lu, 2013, Initial neurite outgrowth in drosophila neurons is driven by kinesin-powered microtubule sliding, Current Biology, 23, 1018, 10.1016/j.cub.2013.04.050

Lu, 2015, Kinesin-1-powered microtubule sliding initiates axonal regeneration in drosophila cultured neurons, Molecular Biology of the Cell, 26, 1296, 10.1091/mbc.E14-10-1423

Luo, 1994, Distinct morphogenetic functions of similar small GTPases: drosophila Drac1 is involved in axonal outgrowth and myoblast fusion, Genes & Development, 8, 1787, 10.1101/gad.8.15.1787

Matov, 2010, Analysis of microtubule dynamic instability using a plus-end growth marker, Nature Methods, 7, 761, 10.1038/nmeth.1493

Mazel, 2014, Direct observation of microtubule pushing by cortical dynein in living cells, Molecular Biology of the Cell, 25, 95, 10.1091/mbc.E13-07-0376

McNally, 1993, Identification of katanin, an ATPase that severs and disassembles stable microtubules, Cell, 75, 419, 10.1016/0092-8674(93)90377-3

Nadar, 2008, Kinesin-5 is essential for growth-cone turning, Current Biology, 18, 1972, 10.1016/j.cub.2008.11.021

Nadar, 2012, Microtubule redistribution in growth cones elicited by focal inactivation of kinesin-5, Journal of Neuroscience, 32, 5783, 10.1523/JNEUROSCI.0144-12.2012

Rogers, 2008, Culture of drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy, Nature Protocols, 3, 606, 10.1038/nprot.2008.18

Rolls, 2007, Polarity and intracellular compartmentalization of drosophila neurons, Neural Development, 2, 7, 10.1186/1749-8104-2-7

Roossien, 2014, Cytoplasmic dynein pushes the cytoskeletal meshwork forward during axonal elongation, Journal of Cell Science, 127, 3593, 10.1242/jcs.152611

Seeger, 2010, Microtubule-associated protein-like binding of the kinesin-1 tail to microtubules, Journal of Biological Chemistry, 285, 8155, 10.1074/jbc.M109.068247

Sharp, 2000, Functional coordination of three mitotic motors in drosophila embryos, Molecular Biology of the Cell, 11, 241, 10.1091/mbc.11.1.241

Shimada, 2006, Polarized transport of frizzled along the planar microtubule arrays in drosophila wing epithelium, Developmental Cell, 10, 209, 10.1016/j.devcel.2005.11.016

Stepanova, 2003, Visualization of microtubule growth in cultured neurons via the use of EB3-GFP (end-binding protein 3-green fluorescent protein), Journal of Neuroscience, 23, 2655, 10.1523/JNEUROSCI.23-07-02655.2003

Stone, 2008, Microtubules have opposite orientation in axons and dendrites of drosophila neurons, Molecular Biology of the Cell, 19, 4122, 10.1091/mbc.E07-10-1079

Straube, 2006, Conventional kinesin mediates microtubule-microtubule interactions in vivo, Molecular Biology of the Cell, 17, 907, 10.1091/mbc.E05-06-0542

Suter, 2011, The emerging role of forces in axonal elongation, Progress in Neurobiology, 94, 91, 10.1016/j.pneurobio.2011.04.002

Tanaka, 2012, Nezha/CAMSAP3 and CAMSAP2 cooperate in epithelial-specific organization of noncentrosomal microtubules, Proceedings of the National Academy of Sciences of the United States of America, 109, 20029, 10.1073/pnas.1218017109

Wang, 2013, Patronin mediates a switch from kinesin-13-dependent poleward flux to anaphase b spindle elongation, The Journal of Cell Biology, 203, 35, 10.1083/jcb.201306001

Yan, 2013, Kinesin-1 regulates dendrite microtubule polarity in caenorhabditis elegans, eLife, 2, e00133, 10.7554/eLife.00133

Zheng, 2008, Dynein is required for polarized dendritic transport and uniform microtubule orientation in axons, Nature Cell Biology, 10, 1172, 10.1038/ncb1777

Thông tin
Thông tin xuất bản

eLife

Tập 4

Thông tin tác giả