Abstract
The distinct electrical properties of axonal and dendritic membranes are largely a result of specific transport of vesicle-bound membrane proteins to each compartment. How this specificity arises is unclear because kinesin motors that transport vesicles cannot autonomously distinguish dendritically projecting microtubules from those projecting axonally. We hypothesized that interaction with a second motor might enable vesicles containing dendritic proteins to preferentially associate with dendritically projecting microtubules and avoid those that project to the axon. Here we show that in rat cortical neurons, localization of several distinct transmembrane proteins to dendrites is dependent on specific myosin motors and an intact actin network. Moreover, fusion with a myosin-binding domain from Melanophilin targeted Channelrhodopsin-2 specifically to the somatodendritic compartment of neurons in mice in vivo. Together, our results suggest that dendritic transmembrane proteins direct the vesicles in which they are transported to avoid the axonal compartment through interaction with myosin motors.
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References
Mostov, K., Su, T. & ter Beest, M. Polarized epithelial membrane traffic: conservation and plasticity. Nat. Cell Biol. 5, 287–293 (2003).
Folsch, H., Ohno, H., Bonifacino, J.S. & Mellman, I. A novel clathrin adaptor complex mediates basolateral targeting in polarized epithelial cells. Cell 99, 189–198 (1999).
Burack, M.A., Silverman, M.A. & Banker, G. The role of selective transport in neuronal protein sorting. Neuron 26, 465–472 (2000).
Matsuda, S. et al. Accumulation of AMPA receptors in autophagosomes in neuronal axons lacking adaptor protein AP-4. Neuron 57, 730–745 (2008).
Setou, M. et al. Glutamate-receptor-interacting protein GRIP1 directly steers kinesin to dendrites. Nature 417, 83–87 (2002).
Setou, M., Nakagawa, T., Seog, D.H. & Hirokawa, N. Kinesin superfamily motor protein KIF17 and mLin-10 in NMDA receptor-containing vesicle transport. Science 288, 1796–1802 (2000).
Chu, P.J., Rivera, J.F. & Arnold, D.B. A role for Kif17 in transport of Kv4.2. J. Biol. Chem. 281, 365–373 (2006).
Nakata, T. & Hirokawa, N. Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head. J. Cell Biol. 162, 1045–1055 (2003).
Kaech, S., Fischer, M., Doll, T. & Matus, A. Isoform specificity in the relationship of actin to dendritic spines. J. Neurosci. 17, 9565–9572 (1997).
Nakata, T., Terada, S. & Hirokawa, N. Visualization of the dynamics of synaptic vesicle and plasma membrane proteins in living axons. J. Cell Biol. 140, 659–674 (1998).
Correia, S.S. et al. Motor protein-dependent transport of AMPA receptors into spines during long-term potentiation. Nat. Neurosci. 11, 457–466 (2008).
Osterweil, E., Wells, D.G. & Mooseker, M.S. A role for myosin VI in postsynaptic structure and glutamate receptor endocytosis. J. Cell Biol. 168, 329–338 (2005).
Wang, Z. et al. Myosin Vb mobilizes recycling endosomes and AMPA receptors for postsynaptic plasticity. Cell 135, 535–548 (2008).
Cheney, R.E. et al. Brain myosin-V is a two-headed unconventional myosin with motor activity. Cell 75, 13–23 (1993).
Lise, M.F. et al. Involvement of myosin Vb in glutamate receptor trafficking. J. Biol. Chem. 281, 3669–3678 (2006).
Ruberti, F. & Dotti, C.G. Involvement of the proximal C terminus of the AMPA receptor subunit GluR1 in dendritic sorting. J. Neurosci. 20, RC78:1–5 (2000).
Mercer, J.A., Seperack, P.K., Strobel, M.C., Copeland, N.G. & Jenkins, N.A. Novel myosin heavy chain encoded by murine dilute coat colour locus. Nature 349, 709–713 (1991).
Sheng, M., Tsaur, M.L., Jan, Y.N. & Jan, L.Y. Subcellular segregation of two A-type K+ channel proteins in rat central neurons. Neuron 9, 271–284 (1992).
Cheng, C., Glover, G., Banker, G. & Amara, S.G. A novel sorting motif in the glutamate transporter excitatory amino acid transporter 3 directs its targeting in Madin-Darby canine kidney cells and hippocampal neurons. J. Neurosci. 22, 10643–10652 (2002).
Das, S.S. & Banker, G.A. The role of protein interaction motifs in regulating the polarity and clustering of the metabotropic glutamate receptor mGluR1a. J. Neurosci. 26, 8115–8125 (2006).
Geething, N.C. & Spudich, J.A. Identification of a minimal myosin Va binding site within an intrinsically unstructured domain of melanophilin. J. Biol. Chem. 282, 21518–21528 (2007).
West, A.E., Neve, R.L. & Buckley, K.M. Identification of a somatodendritic targeting signal in the cytoplasmic domain of the transferrin receptor. J. Neurosci. 17, 6038–6047 (1997).
Rosales, C.R., Osborne, K.D., Zuccarino, G.V., Scheiffele, P. & Silverman, M.A. A cytoplasmic motif targets neuroligin-1 exclusively to dendrites of cultured hippocampal neurons. Eur. J. Neurosci. 22, 2381–2386 (2005).
Bradke, F. & Dotti, C.G. Differentiated neurons retain the capacity to generate axons from dendrites. Curr. Biol. 10, 1467–1470 (2000).
Winckler, B., Forscher, P. & Mellman, I. A diffusion barrier maintains distribution of membrane proteins in polarized neurons. Nature 397, 698–701 (1999).
Passafaro, M. et al. Subunit-specific temporal and spatial patterns of AMPA receptor exocytosis in hippocampal neurons. Nat. Neurosci. 4, 917–926 (2001).
Nagel, G. et al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc. Natl. Acad. Sci. USA 100, 13940–13945 (2003).
Boyden, E.S., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 8, 1263–1268 (2005).
Li, X. et al. Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin. Proc. Natl. Acad. Sci. USA 102, 17816–17821 (2005).
Petreanu, L., Huber, D., Sobczyk, A. & Svoboda, K. Channelrhodopsin-2-assisted circuit mapping of long-range callosal projections. Nat. Neurosci. 10, 663–668 (2007).
Arenkiel, B.R. et al. In vivo light-induced activation of neural circuitry in transgenic mice expressing channelrhodopsin-2. Neuron 54, 205–218 (2007).
Morris, R.L. & Hollenbeck, P.J. Axonal transport of mitochondria along microtubules and F-actin in living vertebrate neurons. J. Cell Biol. 131, 1315–1326 (1995).
Rivera, J.F., Ahmad, S., Quick, M.W., Liman, E.R. & Arnold, D.B. An evolutionarily conserved dileucine motif in Shal K+ channels mediates dendritic targeting. Nat. Neurosci. 6, 243–250 (2003).
Kaether, C., Skehel, P. & Dotti, C.G. Axonal membrane proteins are transported in distinct carriers: a two-color video microscopy study in cultured hippocampal neurons. Mol. Biol. Cell 11, 1213–1224 (2000).
Guillaud, L., Setou, M. & Hirokawa, N. KIF17 dynamics and regulation of NR2B trafficking in hippocampal neurons. J. Neurosci. 23, 131–140 (2003).
Pollard, T.D., Selden, S.C. & Maupin, P. Interaction of actin filaments with microtubules. J. Cell Biol. 99, 33s–37s (1984).
Dehmelt, L. & Halpain, S. Actin and microtubules in neurite initiation: are MAPs the missing link? J. Neurobiol. 58, 18–33 (2004).
Ali, M.Y. et al. Myosin Va maneuvers through actin intersections and diffuses along microtubules. Proc. Natl. Acad. Sci. USA 104, 4332–4336 (2007).
Kural, C. et al. Tracking melanosomes inside a cell to study molecular motors and their interaction. Proc. Natl. Acad. Sci. USA 104, 5378–5382 (2007).
Bridgman, P.C. Myosin Va movements in normal and dilute-lethal axons provide support for a dual filament motor complex. J. Cell Biol. 146, 1045–1060 (1999).
Baas, P.W., Deitch, J.S., Black, M.M. & Banker, G.A. Polarity orientation of microtubules in hippocampal neurons: uniformity in the axon and nonuniformity in the dendrite. Proc. Natl. Acad. Sci. USA 85, 8335–8339 (1988).
Dillman, J. F. 3rd, Dabney, L.P. & Pfister, K.K. Cytoplasmic dynein is associated with slow axonal transport. Proc. Natl. Acad. Sci. USA 93, 141–144 (1996).
Li, X.D. et al. The globular tail domain puts on the brake to stop the ATPase cycle of myosin Va. Proc. Natl. Acad. Sci. USA 105, 1140–1145 (2008).
Pranchevicius, M.C. et al. Myosin Va phosphorylated on Ser1650 is found in nuclear speckles and redistributes to nucleoli upon inhibition of transcription. Cell Motil. Cytoskeleton 65, 441–456 (2008).
Krementsov, D.N., Krementsova, E.B. & Trybus, K.M. Myosin V: regulation by calcium, calmodulin, and the tail domain. J. Cell Biol. 164, 877–886 (2004).
Luo, L., Callaway, E.M. & Svoboda, K. Genetic dissection of neural circuits. Neuron 57, 634–660 (2008).
Huber, D. et al. Sparse optical microstimulation in barrel cortex drives learned behaviour in freely moving mice. Nature 451, 61–64 (2008).
Acknowledgements
The TfR-GFP and JPA5-CD8 (G. Banker, Oregon Health Sciences University), NLG-HA (M. Silverman, Simon Fraser University), GluR1-GFP (R. Malinow, University of California at San Diego) and siRNA vector (J. Esteban, University of Michigan) plasmids and the antibodies to GluR1 (R. Wenthold, US National Institutes of Health–National Institute on Deafness and Other Communication Disorders) and EAAT3 (S. Amara, University of Pittsburgh) were gifts. The authors would like to thank E. Liman, N. Segil, D. McKemy and members of the Arnold lab for comments on the manuscript and S.H. Kwon for creating the illustrations. This work was supported by US National Institutes of Health grants NS-041963 and MH-071439 to D.B.A. and support from the Howard Hughes Medical Institute to K.S.
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D.B.A. and T.L.L. designed and T.L.L. generated chimeric and mutant constructs. D.B.A. and T.L.L. designed and T.L.L. performed experiments to test constructs in dissociated neuronal cultures; K.S. and T.M. designed and performed the slice experiments; D.B.A., T.L.L., T.M. and K.S. analyzed results; D.B.A., T.L.L. and K.S. wrote the paper; and D.B.A. supervised the project.
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Lewis, T., Mao, T., Svoboda, K. et al. Myosin-dependent targeting of transmembrane proteins to neuronal dendrites. Nat Neurosci 12, 568–576 (2009). https://doi.org/10.1038/nn.2318
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DOI: https://doi.org/10.1038/nn.2318
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