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Specific Sorting and Post-Golgi Trafficking of Dendritic Potassium Channels in Living Neurons*

Open AccessPublished:February 25, 2014DOI:https://doi.org/10.1074/jbc.M113.534495
      Proper membrane localization of ion channels is essential for the function of neuronal cells. Particularly, the computational ability of dendrites depends on the localization of different ion channels in specific subcompartments. However, the molecular mechanisms that control ion channel localization in distinct dendritic subcompartments are largely unknown. Here, we developed a quantitative live cell imaging method to analyze protein sorting and post-Golgi vesicular trafficking. We focused on two dendritic voltage-gated potassium channels that exhibit distinct localizations: Kv2.1 in proximal dendrites and Kv4.2 in distal dendrites. Our results show that Kv2.1 and Kv4.2 channels are sorted into two distinct populations of vesicles at the Golgi apparatus. The targeting of Kv2.1 and Kv4.2 vesicles occurred by distinct mechanisms as evidenced by their requirement for specific peptide motifs, cytoskeletal elements, and motor proteins. By live cell and super-resolution imaging, we identified a novel trafficking machinery important for the localization of Kv2.1 channels. Particularly, we identified non-muscle myosin II as an important factor in Kv2.1 trafficking. These findings reveal that the sorting of ion channels at the Golgi apparatus and their subsequent trafficking by unique molecular mechanisms are crucial for their specific localizations within dendrites.

      Introduction

      The somatodendritic compartment of neurons receives and integrates incoming synaptic information. These physiological processes are underpinned by a number of receptors and voltage-gated ion channels that are targeted specifically to this compartment. The highly controlled expression of receptors and ion channels and their accurate positioning within the dendrites are therefore important for dendritic computation. However, despite this importance, the mechanisms underlying intracellular trafficking and precise targeting of these molecules remain poorly understood.
      Previous studies suggest that membrane proteins localized in dendrites are selectively delivered from the soma to the dendrites by vesicular transport (
      • Burack M.A.
      • Silverman M.A.
      • Banker G.
      The role of selective transport in neuronal protein sorting.
      ,
      • Sampo B.
      • Kaech S.
      • Kunz S.
      • Banker G.
      Two distinct mechanisms target membrane proteins to the axonal surface.
      ) and are prevented from entering the axon at the axon initial segment (
      • Al-Bassam S.
      • Xu M.
      • Wandless T.J.
      • Arnold D.B.
      Differential trafficking of transport vesicles contributes to the localization of dendritic proteins.
      ,
      • Watanabe K.
      • Al-Bassam S.
      • Miyazaki Y.
      • Wandless T.J.
      • Webster P.
      • Arnold D.B.
      Networks of polarized actin filaments in the axon initial segment provide a mechanism for sorting axonal and dendritic proteins.
      ). However, it is still uncertain how receptors and ion channels are targeted to more confined areas within the dendrites, such as proximal dendrites, distal dendrites, and dendritic spines (
      • Lai H.C.
      • Jan L.Y.
      The distribution and targeting of neuronal voltage-gated ion channels.
      ).
      In the present study, we hypothesized that, within dendrites, multiple biosynthetic trafficking mechanisms exist to target membrane proteins to different and specific subcompartments. To test this hypothesis, we focused on Kv2.1
      The abbreviations used are: Kv
      Voltage-gated K+
      DIV
      days in vitro
      MyoIIB
      non-muscle myosin IIB
      CCD
      charge-coupled device
      PALM
      photoactivated localization microscopy
      PA-TagRFP
      photoactivatable red fluorescent protein
      AIS
      axon initial segment
      LatA
      latrunculin A
      MAP-2
      microtubule-associated protein-2.
      and Kv4.2 channels based on their distinct localizations in dendrites. Kv2.1 channels are widely expressed in the mammalian brain where they exhibit restricted localization on soma and proximal parts of the dendrites (
      • Murakoshi H.
      • Trimmer J.S.
      Identification of the Kv2.1 K+ channel as a major component of the delayed rectifier K+ current in rat hippocampal neurons.
      ). In contrast, Kv4.2 channels are expressed in distal parts of the dendrites with increasing expression levels from the soma (
      • Menegola M.
      • Misonou H.
      • Vacher H.
      • Trimmer J.S.
      Dendritic A-type potassium channel subunit expression in CA1 hippocampal interneurons.
      ). These Kv channels, therefore, serve as excellent model proteins to study targeting to different subcompartments of neuronal dendrites (
      • Jensen C.S.
      • Rasmussen H.B.
      • Misonou H.
      Neuronal trafficking of voltage-gated potassium channels.
      ).
      We first developed quantitative live cell imaging methods to investigate whether different ion channels are sorted into distinct transport vesicles at the Golgi apparatus. Using Kv channels tagged with fluorescent proteins, we here demonstrate a novel approach to synchronize the release of transport vesicles from the Golgi apparatus by a modified temperature arrest method. With this approach, we uncovered that Kv2.1 and Kv4.2 channels are sorted into different types of transport vesicles, which display different movement dynamics and distributions. The specific sorting into vesicles was found to be dictated by previously reported peptide targeting motifs present in Kv2.1 (
      • Lim S.T.
      • Antonucci D.E.
      • Scannevin R.H.
      • Trimmer J.S.
      A novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons.
      ) and Kv4.2 (
      • 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.
      ). Quantitative analysis revealed that the differential targeting of Kv2.1 and Kv4.2 post-Golgi transport vesicles is mediated by distinct molecular mechanisms. Particularly, we identified a novel actin-myosin-based trafficking mechanism for Kv2.1 of which disruption caused a marked change in the expression of the channel. Our imaging study, therefore, provides insight into how dendritic ion channels are localized to specific subcompartments in neuronal cells. The novel imaging technique introduced here also provides a basis for future investigations of vesicular trafficking of a broader spectrum of membrane signaling proteins in neurons.

      DISCUSSION

      In this study, we developed a novel imaging approach and used it to probe protein sorting and vesicular transport of dendritic Kv channels in living neurons. Our approach, based on the temperature block, resulted in the effective accumulation of GFP-tagged channel proteins in the Golgi apparatus, thereby allowing semipulse-chase imaging of the post-Golgi vesicular trafficking of Kv channels. By combining this approach with a number of quantitative analyses, we provided evidence that two dendritic ion channels, Kv2.1 and Kv4.2, are precisely sorted into distinct pools of transport vesicles that are targeted to specific dendritic subcompartments by different molecular mechanisms. Previous work has demonstrated that membrane proteins targeted to either axonal or somatodendritic compartments, such as the transferrin receptor and neuroglia cell adhesion molecule (NgCAM) are sorted in distinct populations of transport vesicles (
      • Burack M.A.
      • Silverman M.A.
      • Banker G.
      The role of selective transport in neuronal protein sorting.
      ), a result that has also been supported by recent studies (
      • Lewis Jr., T.L.
      • Mao T.
      • Svoboda K.
      • Arnold D.B.
      Myosin-dependent targeting of transmembrane proteins to neuronal dendrites.
      ,
      • Song A.H.
      • Wang D.
      • Chen G.
      • Li Y.
      • Luo J.
      • Duan S.
      • Poo M.M.
      A selective filter for cytoplasmic transport at the axon initial segment.
      ). Our study further extends this compartment-specific trafficking model to ion channels that are localized to specific subcompartments within the dendrites. Based on our findings, we propose a selective dendritic transport model (Fig. 11E) in which proximal Kv2.1 and distal Kv4.2 are sorted into unique populations of vesicles at the Golgi apparatus and delivered by distinct transport machineries.
      The significance of the specific sorting in establishing the distinct localizations of Kv2.1 and Kv4.2 is supported by our experiments using localization mutants. We showed that the mutants, which have been reported to mislocalize in neuronal membranes (
      • Lim S.T.
      • Antonucci D.E.
      • Scannevin R.H.
      • Trimmer J.S.
      A novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons.
      ,
      • 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.
      ), were indeed impaired in the protein sorting process. They appeared to be still efficiently packaged into post-Golgi transport vesicles but not into their designated populations. Consequently, these vesicles carrying missorted proteins were transported to improper locations, such as the distal dendrites for Kv2.1 S586A and the axon for Kv4.2 LL/AV. These results indicate that sorting to a specific population of transport vesicles at the Golgi apparatus is a crucial step in localizing these Kv channels to specific dendritic subcompartments.
      Interestingly, our studies also shed light on how efficiently and precisely neurons sort dendritic ion channels into post-Golgi vesicles. Ample evidence suggests that the binding of a specific set of coat proteins to a peptide sorting signal is the common mechanism whereby membrane proteins are concentrated and packaged into a specific population of vesicles at the Golgi apparatus (
      • Bonifacino J.S.
      • Lippincott-Schwartz J.
      Coat proteins: shaping membrane transport.
      ,
      • Traub L.M.
      Common principles in clathrin-mediated sorting at the Golgi and the plasma membrane.
      ). A recent study also reported that the μ1A subunit of adaptor protein-1 complex prevents somatodendritic proteins from being packaged into axonal transport vesicles at the Golgi apparatus (
      • Farías G.G.
      • Cuitino L.
      • Guo X.
      • Ren X.
      • Jarnik M.
      • Mattera R.
      • Bonifacino J.S.
      Signal-mediated, AP-1/clathrin-dependent sorting of transmembrane receptors to the somatodendritic domain of hippocampal neurons.
      ). Although we currently do not know the molecular mechanism whereby dendritic Kv2.1 and Kv4.2 are separated into specific vesicular carriers, our results that an Arf1 mutant inhibits the formation of both types of vesicles indicate that adaptor protein-1 or other clathrin-associated sorting proteins recruited by Arf1 (
      • Traub L.M.
      Common principles in clathrin-mediated sorting at the Golgi and the plasma membrane.
      ,
      • Edeling M.A.
      • Smith C.
      • Owen D.
      Life of a clathrin coat: insights from clathrin and AP structures.
      ) might be involved in the sorting events.
      The velocities of post-Golgi transport vesicles obtained in this study (0.5–6 μm/s) are much greater than the values reported for single myosin IIB molecules (<0.1 μm/s) (
      • Norstrom M.F.
      • Smithback P.A.
      • Rock R.S.
      Unconventional processive mechanics of non-muscle myosin IIB.
      ) and kinesin motors (up to 2 μm/s) (
      • Hirokawa N.
      • Noda Y.
      • Tanaka Y.
      • Niwa S.
      Kinesin superfamily motor proteins and intracellular transport.
      ), which have been measured mostly in vitro. As our results with vesicular stomatitis virus G-GFP showed velocities comparable with those measured previously in living cells (
      • Hirschberg K.
      • Miller C.M.
      • Ellenberg J.
      • Presley J.F.
      • Siggia E.D.
      • Phair R.D.
      • Lippincott-Schwartz J.
      Kinetic analysis of secretory protein traffic and characterization of Golgi to plasma membrane transport intermediates in living cells.
      ,
      • Toomre D.
      • Keller P.
      • White J.
      • Olivo J.C.
      • Simons K.
      Dual-color visualization of trans-Golgi network to plasma membrane traffic along microtubules in living cells.
      ,
      • Polishchuk E.V.
      • Di Pentima A.
      • Luini A.
      • Polishchuk R.S.
      Mechanism of constitutive export from the Golgi: bulk flow via the formation, protrusion, and en bloc cleavage of large trans-Golgi network tubular domains.
      • Lock J.G.
      • Stow J.L.
      Rab11 in recycling endosomes regulates the sorting and basolateral transport of E-cadherin.
      ), we concluded that our measurements are genuine. It should also be noted that the most classical type of vesicular transport in neurons, the fast axonal transport, has been shown to operate at the speed of ∼5 μm/s (400 mm/day). Therefore, there may be unknown factors that accelerate molecular motors, presumably their hydrolysis cycle (
      • Howard J.
      Mechanics of Motor Proteins and the Cytoskeleton.
      ), in the native cellular environment. However, these remain to be identified.
      Previous studies have characterized mobile structures positive for Kv2.1 in neurons using live cell imaging (
      • Nakata T.
      • Hirokawa N.
      Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head.
      ,
      • Tamkun M.M.
      • O'connell K.M.
      • Rolig A.S.
      A cytoskeletal-based perimeter fence selectively corrals a sub-population of cell surface Kv2.1 channels.
      ). However, because vesicles mediate the transport of membrane proteins between several compartments, such as from endoplasmic reticulum to Golgi or from the surface to endosomes, simple live cell imaging does not provide direct insight into the post-Golgi trafficking of Kv2.1. Our approach combining temperature block with quantitative live cell imaging allowed us to detect and analyze post-Golgi transport vesicles preferentially. Here, we provided evidence that GFP-Kv2.1 is sorted into a specific pool of vesicles at the Golgi apparatus that differs from the vesicle population carrying GFP-Kv4.2. Furthermore, GFP-Kv2.1 vesicles were trafficked by a unique mechanism involving myosin IIB and actin filaments. The molecular interaction detected between endogenous Kv2.1 and myosin IIB in brain tissue suggests that this novel trafficking mechanism identified for GFP-Kv2.1 also applies to the native channel. This was further supported by the results that shRNAs for myosin IIB caused significant changes in the expression of endogenous Kv2.1.
      The binding between Kv2.1 and myosin IIB appears to be direct. This was evidenced by the results that recombinant myosin IIB expressed in E. coli was able to bind to recombinant cytoplasmic fragments of Kv2.1. This is comparable with the direct binding of the KIF17 motor protein to Kv4.2 (
      • Chu P.J.
      • Rivera J.F.
      • Arnold D.B.
      A role for Kif17 in transport of Kv4.2.
      ). Also, the binding may involve both cytoplasmic tails of Kv2.1 as myosin IIB was co-purified with both of the fragments. This is interesting in light of the previous work demonstrating that the N and C termini of Kv2.1 can form a polypeptide complex and facilitate Kv2.1 trafficking in heterologous cells (
      • Mohapatra D.P.
      • Siino D.F.
      • Trimmer J.S.
      Interdomain cytoplasmic interactions govern the intracellular trafficking, gating, and modulation of the Kv2.1 channel.
      ).
      It has been shown that Kv2.1 is also localized in the proximal region of the axon, the AIS (
      • Lim S.T.
      • Antonucci D.E.
      • Scannevin R.H.
      • Trimmer J.S.
      A novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons.
      ,
      • Sarmiere P.D.
      • Weigle C.M.
      • Tamkun M.M.
      The Kv2.1 K+ channel targets to the axon initial segment of hippocampal and cortical neurons in culture and in situ.
      ). Although the S586A mutation disrupted the proximal localization of Kv2.1 in dendrites, it did not affect its restricted localization at the AIS (see Fig. 3A) (
      • Lim S.T.
      • Antonucci D.E.
      • Scannevin R.H.
      • Trimmer J.S.
      A novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons.
      ). This suggests that the mechanism that localizes Kv2.1 to the AIS differs from the mechanism mediating the restricted somatodendritic localization of Kv2.1.
      Our live PALM study provided evidence that actin filaments are oriented randomly in proximal dendrites. This is consistent with the random motion of Kv2.1 vesicles presumably mediated by a myosin motor. Although we do not know exactly how long actin filaments are, actin filament length in neurons varies widely, ranging from a few micrometers to 100 nm (
      • Xu K.
      • Zhong G.
      • Zhuang X.
      Actin, spectrin, and associated proteins form a periodic cytoskeletal structure in axons.
      ,
      • Korobova F.
      • Svitkina T.
      Molecular architecture of synaptic actin cytoskeleton in hippocampal neurons reveals a mechanism of dendritic spine morphogenesis.
      ). Therefore, motor molecules (and Kv2.1 vesicles) may have to change tracks frequently in the proximal dendrites. The random orientation and the short track length would cause frequent stalls and changes of direction of Kv2.1 transport vesicles in accordance with our observation. Alternatively, the myosin motor itself may reverse direction as observed by single molecule imaging of myosin IIB (
      • Norstrom M.F.
      • Smithback P.A.
      • Rock R.S.
      Unconventional processive mechanics of non-muscle myosin IIB.
      ).
      In summary, using quantitative imaging approaches, we provide evidence that neurons sort dendritic ion channels into multiple trafficking pathways, thereby resulting in destination-specific targeting and localization. In addition, these approaches would be useful in future studies to determine vesicular trafficking of other membrane signaling molecules in living neurons.

      Acknowledgments

      We thank Drs. James Trimmer, Stephanie Kaech, Mark Rizzo, Takashi Tsuboi, and Don Arnold for generously providing precious reagents. We also thank Matthew Rasband and Scott Thompson for critical reading and helpful discussion.

      REFERENCES

        • Burack M.A.
        • Silverman M.A.
        • Banker G.
        The role of selective transport in neuronal protein sorting.
        Neuron. 2000; 26: 465-472
        • Sampo B.
        • Kaech S.
        • Kunz S.
        • Banker G.
        Two distinct mechanisms target membrane proteins to the axonal surface.
        Neuron. 2003; 37: 611-624
        • Al-Bassam S.
        • Xu M.
        • Wandless T.J.
        • Arnold D.B.
        Differential trafficking of transport vesicles contributes to the localization of dendritic proteins.
        Cell Rep. 2012; 2: 89-100
        • Watanabe K.
        • Al-Bassam S.
        • Miyazaki Y.
        • Wandless T.J.
        • Webster P.
        • Arnold D.B.
        Networks of polarized actin filaments in the axon initial segment provide a mechanism for sorting axonal and dendritic proteins.
        Cell Rep. 2012; 2: 1546-1553
        • Lai H.C.
        • Jan L.Y.
        The distribution and targeting of neuronal voltage-gated ion channels.
        Nat. Rev. Neurosci. 2006; 7: 548-562
        • Murakoshi H.
        • Trimmer J.S.
        Identification of the Kv2.1 K+ channel as a major component of the delayed rectifier K+ current in rat hippocampal neurons.
        J. Neurosci. 1999; 19: 1728-1735
        • Menegola M.
        • Misonou H.
        • Vacher H.
        • Trimmer J.S.
        Dendritic A-type potassium channel subunit expression in CA1 hippocampal interneurons.
        Neuroscience. 2008; 154: 953-964
        • Jensen C.S.
        • Rasmussen H.B.
        • Misonou H.
        Neuronal trafficking of voltage-gated potassium channels.
        Mol. Cell. Neurosci. 2011; 48: 288-297
        • Lim S.T.
        • Antonucci D.E.
        • Scannevin R.H.
        • Trimmer J.S.
        A novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons.
        Neuron. 2000; 25: 385-397
        • 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. 2003; 6: 243-250
        • Kaech S.
        • Ludin B.
        • Matus A.
        Cytoskeletal plasticity in cells expressing neuronal microtubule-associated proteins.
        Neuron. 1996; 17: 1189-1199
        • Gilling M.
        • Rasmussen H.B.
        • Calloe K.
        • Sequeira A.F.
        • Baretto M.
        • Oliveira G.
        • Almeida J.
        • Lauritsen M.B.
        • Ullmann R.
        • Boonen S.E.
        • Brondum-Nielsen K.
        • Kalscheuer V.M.
        • Tümer Z.
        • Vicente A.M.
        • Schmitt N.
        • Tommerup N.
        Dysfunction of the heteromeric KV7.3/KV7.5 potassium channel is associated with autism spectrum disorders.
        Front. Genet. 2013; 4: 54
        • Kaech S.
        • Banker G.
        Culturing hippocampal neurons.
        Nat. Protoc. 2006; 1: 2406-2415
        • Shibata R.
        • Misonou H.
        • Campomanes C.R.
        • Anderson A.E.
        • Schrader L.A.
        • Doliveira L.C.
        • Carroll K.I.
        • Sweatt J.D.
        • Rhodes K.J.
        • Trimmer J.S.
        A fundamental role for KChIPs in determining the molecular properties and trafficking of Kv4.2 potassium channels.
        J. Biol. Chem. 2003; 278: 36445-36454
        • Trimmer J.S.
        Immunological identification and characterization of a delayed rectifier K+ channel polypeptide in rat brain.
        Proc. Natl. Acad. Sci. U.S.A. 1991; 88: 10764-10768
        • Frost N.A.
        • Shroff H.
        • Kong H.
        • Betzig E.
        • Blanpied T.A.
        Single-molecule discrimination of discrete perisynaptic and distributed sites of actin filament assembly within dendritic spines.
        Neuron. 2010; 67: 86-99
        • Manley S.
        • Gillette J.M.
        • Patterson G.H.
        • Shroff H.
        • Hess H.F.
        • Betzig E.
        • Lippincott-Schwartz J.
        High-density mapping of single-molecule trajectories with photoactivated localization microscopy.
        Nat. Methods. 2008; 5: 155-157
        • An W.F.
        • Bowlby M.R.
        • Betty M.
        • Cao J.
        • Ling H.P.
        • Mendoza G.
        • Hinson J.W.
        • Mattsson K.I.
        • Strassle B.W.
        • Trimmer J.S.
        • Rhodes K.J.
        Modulation of A-type potassium channels by a family of calcium sensors.
        Nature. 2000; 403: 553-556
        • Frech G.C.
        • VanDongen A.M.
        • Schuster G.
        • Brown A.M.
        • Joho R.H.
        A novel potassium channel with delayed rectifier properties isolated from rat brain by expression cloning.
        Nature. 1989; 340: 642-645
        • Dascher C.
        • Balch W.E.
        Dominant inhibitory mutants of ARF1 block endoplasmic reticulum to Golgi transport and trigger disassembly of the Golgi apparatus.
        J. Biol. Chem. 1994; 269: 1437-1448
        • Jeyifous O.
        • Waites C.L.
        • Specht C.G.
        • Fujisawa S.
        • Schubert M.
        • Lin E.I.
        • Marshall J.
        • Aoki C.
        • de Silva T.
        • Montgomery J.M.
        • Garner C.C.
        • Green W.N.
        SAP97 and CASK mediate sorting of NMDA receptors through a previously unknown secretory pathway.
        Nat. Neurosci. 2009; 12: 1011-1019
        • Hirschberg K.
        • Miller C.M.
        • Ellenberg J.
        • Presley J.F.
        • Siggia E.D.
        • Phair R.D.
        • Lippincott-Schwartz J.
        Kinetic analysis of secretory protein traffic and characterization of Golgi to plasma membrane transport intermediates in living cells.
        J. Cell Biol. 1998; 143: 1485-1503
        • Toomre D.
        • Keller P.
        • White J.
        • Olivo J.C.
        • Simons K.
        Dual-color visualization of trans-Golgi network to plasma membrane traffic along microtubules in living cells.
        J. Cell Sci. 1999; 112: 21-33
        • Polishchuk E.V.
        • Di Pentima A.
        • Luini A.
        • Polishchuk R.S.
        Mechanism of constitutive export from the Golgi: bulk flow via the formation, protrusion, and en bloc cleavage of large trans-Golgi network tubular domains.
        Mol. Biol. Cell. 2003; 14: 4470-4485
        • Lock J.G.
        • Stow J.L.
        Rab11 in recycling endosomes regulates the sorting and basolateral transport of E-cadherin.
        Mol. Biol. Cell. 2005; 16: 1744-1755
        • Sarmiere P.D.
        • Weigle C.M.
        • Tamkun M.M.
        The Kv2.1 K+ channel targets to the axon initial segment of hippocampal and cortical neurons in culture and in situ.
        BMC Neurosci. 2008; 9: 112
        • Brown J.R.
        • Stafford P.
        • Langford G.M.
        Short-range axonal/dendritic transport by myosin-V: a model for vesicle delivery to the synapse.
        J. Neurobiol. 2004; 58: 175-188
        • Osterweil E.
        • Wells D.G.
        • Mooseker M.S.
        A role for myosin VI in postsynaptic structure and glutamate receptor endocytosis.
        J. Cell Biol. 2005; 168: 329-338
        • Chu P.J.
        • Rivera J.F.
        • Arnold D.B.
        A role for Kif17 in transport of Kv4.2.
        J. Biol. Chem. 2006; 281: 365-373
        • Xu K.
        • Zhong G.
        • Zhuang X.
        Actin, spectrin, and associated proteins form a periodic cytoskeletal structure in axons.
        Science. 2013; 339: 452-456
        • Betzig E.
        • Patterson G.H.
        • Sougrat R.
        • Lindwasser O.W.
        • Olenych S.
        • Bonifacino J.S.
        • Davidson M.W.
        • Lippincott-Schwartz J.
        • Hess H.F.
        Imaging intracellular fluorescent proteins at nanometer resolution.
        Science. 2006; 313: 1642-1645
        • Pollard T.D.
        • Borisy G.G.
        Cellular motility driven by assembly and disassembly of actin filaments.
        Cell. 2003; 112: 453-465
        • Subach F.V.
        • Patterson G.H.
        • Renz M.
        • Lippincott-Schwartz J.
        • Verkhusha V.V.
        Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells.
        J. Am. Chem. Soc. 2010; 132: 6481-6491
        • Straight A.F.
        • Cheung A.
        • Limouze J.
        • Chen I.
        • Westwood N.J.
        • Sellers J.R.
        • Mitchison T.J.
        Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor.
        Science. 2003; 299: 1743-1747
        • Allingham J.S.
        • Smith R.
        • Rayment I.
        The structural basis of blebbistatin inhibition and specificity for myosin II.
        Nat. Struct. Mol. Biol. 2005; 12: 378-379
        • Los G.V.
        • Encell L.P.
        • McDougall M.G.
        • Hartzell D.D.
        • Karassina N.
        • Zimprich C.
        • Wood M.G.
        • Learish R.
        • Ohana R.F.
        • Urh M.
        • Simpson D.
        • Mendez J.
        • Zimmerman K.
        • Otto P.
        • Vidugiris G.
        • Zhu J.
        • Darzins A.
        • Klaubert D.H.
        • Bulleit R.F.
        • Wood K.V.
        HaloTag: a novel protein labeling technology for cell imaging and protein analysis.
        ACS Chem. Biol. 2008; 3: 373-382
        • Lewis Jr., T.L.
        • Mao T.
        • Svoboda K.
        • Arnold D.B.
        Myosin-dependent targeting of transmembrane proteins to neuronal dendrites.
        Nat. Neurosci. 2009; 12: 568-576
        • Song A.H.
        • Wang D.
        • Chen G.
        • Li Y.
        • Luo J.
        • Duan S.
        • Poo M.M.
        A selective filter for cytoplasmic transport at the axon initial segment.
        Cell. 2009; 136: 1148-1160
        • Bonifacino J.S.
        • Lippincott-Schwartz J.
        Coat proteins: shaping membrane transport.
        Nat. Rev. Mol. Cell Biol. 2003; 4: 409-414
        • Traub L.M.
        Common principles in clathrin-mediated sorting at the Golgi and the plasma membrane.
        Biochim. Biophys. Acta. 2005; 1744: 415-437
        • Farías G.G.
        • Cuitino L.
        • Guo X.
        • Ren X.
        • Jarnik M.
        • Mattera R.
        • Bonifacino J.S.
        Signal-mediated, AP-1/clathrin-dependent sorting of transmembrane receptors to the somatodendritic domain of hippocampal neurons.
        Neuron. 2012; 75: 810-823
        • Edeling M.A.
        • Smith C.
        • Owen D.
        Life of a clathrin coat: insights from clathrin and AP structures.
        Nat. Rev. Mol. Cell Biol. 2006; 7: 32-44
        • Norstrom M.F.
        • Smithback P.A.
        • Rock R.S.
        Unconventional processive mechanics of non-muscle myosin IIB.
        J. Biol. Chem. 2010; 285: 26326-26334
        • Hirokawa N.
        • Noda Y.
        • Tanaka Y.
        • Niwa S.
        Kinesin superfamily motor proteins and intracellular transport.
        Nat. Rev. Mol. Cell Biol. 2009; 10: 682-696
        • Howard J.
        Mechanics of Motor Proteins and the Cytoskeleton.
        Sinauer Associates, Inc., Sunderland, MA2001
        • Nakata T.
        • Hirokawa N.
        Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head.
        J. Cell Biol. 2003; 162: 1045-1055
        • Tamkun M.M.
        • O'connell K.M.
        • Rolig A.S.
        A cytoskeletal-based perimeter fence selectively corrals a sub-population of cell surface Kv2.1 channels.
        J. Cell Sci. 2007; 120: 2413-2423
        • Mohapatra D.P.
        • Siino D.F.
        • Trimmer J.S.
        Interdomain cytoplasmic interactions govern the intracellular trafficking, gating, and modulation of the Kv2.1 channel.
        J. Neurosci. 2008; 28: 4982-4994
        • Korobova F.
        • Svitkina T.
        Molecular architecture of synaptic actin cytoskeleton in hippocampal neurons reveals a mechanism of dendritic spine morphogenesis.
        Mol. Biol. Cell. 2010; 21: 165-176