Formation of elaborately branched dendrites is necessary for the proper input and connectivity of many sensory neurons. Previous studies have revealed that dendritic growth relies heavily on ER-to-Golgi transport, Golgi outposts and endocytic recycling. How new membrane and associated cargo is delivered from the secretory and endosomal compartments to sites of active dendritic growth, however, remains unknown. Using a candidate-based genetic screen in C. elegans, we have identified the small GTPase RAB-10 as a key regulator of membrane trafficking during dendrite morphogenesis. Loss of rab-10 severely reduced proximal dendritic arborization in the multi-dendritic PVD neuron. RAB-10 acts cell-autonomously in the PVD neuron and localizes to the Golgi and early endosomes. Loss of function mutations of the exocyst complex components exoc-8 and sec-8, which regulate tethering, docking and fusion of transport vesicles at the plasma membrane, also caused proximal dendritic arborization defects and led to the accumulation of intracellular RAB-10 vesicles. In rab-10 and exoc-8 mutants, the trans-membrane proteins DMA-1 and HPO-30, which promote PVD dendrite stabilization and branching, no longer localized strongly to the proximal dendritic membranes and instead were sequestered within intracellular vesicles. Together these results suggest a crucial role for the Rab10 GTPase and the exocyst complex in controlling membrane transport from the secretory and/or endosomal compartments that is required for dendritic growth.
References
[1]
Ye B, Zhang Y, Song W, Younger SH, Jan LY, et al. (2007) Growing dendrites and axons differ in their reliance on the secretory pathway. Cell 130: 717–729. pmid:17719548 doi: 10.1016/j.cell.2007.06.032
[2]
Satoh D, Sato D, Tsuyama T, Saito M, Ohkura H, et al. (2008) Spatial control of branching within dendritic arbors by dynein-dependent transport of Rab5-endosomes. Nat Cell Biol 10: 1164–1171. doi: 10.1038/ncb1776. pmid:18758452
[3]
Lee MC, Miller EA, Goldberg J, Orci L, Schekman R (2004) Bi-directional protein transport between the ER and Golgi. Annu Rev Cell Dev Biol 20: 87–123. pmid:15473836 doi: 10.1146/annurev.cellbio.20.010403.105307
[4]
Lazo OM, Gonzalez A, Ascano M, Kuruvilla R, Couve A, et al. (2013) BDNF regulates Rab11-mediated recycling endosome dynamics to induce dendritic branching. J Neurosci 33: 6112–6122. doi: 10.1523/JNEUROSCI.4630-12.2013. pmid:23554492
[5]
Way JC, Chalfie M (1989) The mec-3 gene of Caenorhabditis elegans requires its own product for maintained expression and is expressed in three neuronal cell types. Genes Dev 3: 1823–1833. pmid:2576011 doi: 10.1101/gad.3.12a.1823
[6]
Chatzigeorgiou M, Yoo S, Watson JD, Lee WH, Spencer WC, et al. (2010) Specific roles for DEG/ENaC and TRP channels in touch and thermosensation in C. elegans nociceptors. Nat Neurosci 13: 861–868. doi: 10.1038/nn.2581. pmid:20512132
[7]
Li W, Kang L, Piggott BJ, Feng Z, Xu XZ (2011) The neural circuits and sensory channels mediating harsh touch sensation in Caenorhabditis elegans. Nat Commun 2: 315. doi: 10.1038/ncomms1308. pmid:21587232
[8]
Smith CJ, Watson JD, Spencer WC, O'Brien T, Cha B, et al. (2010) Time-lapse imaging and cell-specific expression profiling reveal dynamic branching and molecular determinants of a multi-dendritic nociceptor in C. elegans. Dev Biol 345: 18–33. doi: 10.1016/j.ydbio.2010.05.502. pmid:20537990
[9]
Liu OW, Shen K (2012) The transmembrane LRR protein DMA-1 promotes dendrite branching and growth in C. elegans. Nat Neurosci 15: 57–63. doi: 10.1038/nn.2978
[10]
Dong X, Liu OW, Howell AS, Shen K (2013) An extracellular adhesion molecule complex patterns dendritic branching and morphogenesis. Cell 155: 296–307. doi: 10.1016/j.cell.2013.08.059. pmid:24120131
[11]
Salzberg Y, Diaz-Balzac CA, Ramirez-Suarez NJ, Attreed M, Tecle E, et al. (2013) Skin-derived cues control arborization of sensory dendrites in Caenorhabditis elegans. Cell 155: 308–320. doi: 10.1016/j.cell.2013.08.058. pmid:24120132
[12]
Smith CJ, O'Brien T, Chatzigeorgiou M, Spencer WC, Feingold-Link E, et al. (2013) Sensory neuron fates are distinguished by a transcriptional switch that regulates dendrite branch stabilization. Neuron 79: 266–280. doi: 10.1016/j.neuron.2013.05.009. pmid:23889932
[13]
Wang T, Liu Y, Xu XH, Deng CY, Wu KY, et al. (2011) Lgl1 activation of rab10 promotes axonal membrane trafficking underlying neuronal polarization. Dev Cell 21: 431–444. doi: 10.1016/j.devcel.2011.07.007. pmid:21856246
[14]
Liu Y, Xu XH, Chen Q, Wang T, Deng CY, et al. (2013) Myosin Vb controls biogenesis of post-Golgi Rab10 carriers during axon development. Nat Commun 4: 2005. doi: 10.1038/ncomms3005. pmid:23770993
[15]
Deng CY, Lei WL, Xu XH, Ju XC, Liu Y, et al. (2014) JIP1 mediates anterograde transport of Rab10 cargos during neuronal polarization. J Neurosci 34: 1710–1723. doi: 10.1523/JNEUROSCI.4496-13.2014. pmid:24478353
[16]
Xu XH, Deng CY, Liu Y, He M, Peng J, et al. (2014) MARCKS regulates membrane targeting of Rab10 vesicles to promote axon development. Cell Res 24: 576–594. doi: 10.1038/cr.2014.33. pmid:24662485
[17]
He B, Guo W (2009) The exocyst complex in polarized exocytosis. Curr Opin Cell Biol 21: 537–542. doi: 10.1016/j.ceb.2009.04.007. pmid:19473826
[18]
Frische EW, Pellis-van Berkel W, van Haaften G, Cuppen E, Plasterk RH, et al. (2007) RAP-1 and the RAL-1/exocyst pathway coordinate hypodermal cell organization in Caenorhabditis elegans. EMBO J 26: 5083–5092. pmid:17989692 doi: 10.1038/sj.emboj.7601922
[19]
Babbey CM, Bacallao RL, Dunn KW (2010) Rab10 associates with primary cilia and the exocyst complex in renal epithelial cells. Am J Physiol Renal Physiol 299: F495–506. doi: 10.1152/ajprenal.00198.2010. pmid:20576682
[20]
Sano H, Eguez L, Teruel MN, Fukuda M, Chuang TD, et al. (2007) Rab10, a target of the AS160 Rab GAP, is required for insulin-stimulated translocation of GLUT4 to the adipocyte plasma membrane. Cell Metab 5: 293–303. pmid:17403373 doi: 10.1016/j.cmet.2007.03.001
[21]
Inoue M, Chang L, Hwang J, Chiang SH, Saltiel AR (2003) The exocyst complex is required for targeting of Glut4 to the plasma membrane by insulin. Nature 422: 629–633. pmid:12687004 doi: 10.1038/nature01533
[22]
Babbey CM, Ahktar N, Wang E, Chen CC, Grant BD, et al. (2006) Rab10 regulates membrane transport through early endosomes of polarized Madin-Darby canine kidney cells. Mol Biol Cell 17: 3156–3175. pmid:16641372 doi: 10.1091/mbc.e05-08-0799
[23]
Schuck S, Gerl MJ, Ang A, Manninen A, Keller P, et al. (2007) Rab10 is involved in basolateral transport in polarized Madin-Darby canine kidney cells. Traffic 8: 47–60. pmid:17132146 doi: 10.1111/j.1600-0854.2006.00506.x
[24]
Chen CC, Schweinsberg PJ, Vashist S, Mareiniss DP, Lambie EJ, et al. (2006) RAB-10 is required for endocytic recycling in the Caenorhabditis elegans intestine. Mol Biol Cell 17: 1286–1297. pmid:16394106 doi: 10.1091/mbc.e05-08-0787
[25]
Glodowski DR, Chen CC, Schaefer H, Grant BD, Rongo C (2007) RAB-10 regulates glutamate receptor recycling in a cholesterol-dependent endocytosis pathway. Mol Biol Cell 18: 4387–4396. pmid:17761527 doi: 10.1091/mbc.e07-05-0486
[26]
Shi A, Chen CC, Banerjee R, Glodowski D, Audhya A, et al. (2010) EHBP-1 functions with RAB-10 during endocytic recycling in Caenorhabditis elegans. Mol Biol Cell 21: 2930–2943. doi: 10.1091/mbc.E10-02-0149. pmid:20573983
[27]
Arakawa H, Kudo H, Batrak V, Caldwell RB, Rieger MA, et al. (2008) Protein evolution by hypermutation and selection in the B cell line DT40. Nucleic Acids Res 36: e1. pmid:18073192 doi: 10.1093/nar/gkm616
[28]
Frokjaer-Jensen C, Davis MW, Sarov M, Taylor J, Flibotte S, et al. (2014) Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon. Nat Methods 11: 529–534. doi: 10.1038/nmeth.2889. pmid:24820376
[29]
Guo P, Hu T, Zhang J, Jiang S, Wang X (2010) Sequential action of Caenorhabditis elegans Rab GTPases regulates phagolysosome formation during apoptotic cell degradation. Proc Natl Acad Sci U S A 107: 18016–18021. doi: 10.1073/pnas.1008946107. pmid:20921409
[30]
Grant B, Zhang Y, Paupard MC, Lin SX, Hall DH, et al. (2001) Evidence that RME-1, a conserved C. elegans EH-domain protein, functions in endocytic recycling. Nat Cell Biol 3: 573–579. pmid:11389442
[31]
Chen B, Jiang Y, Zeng S, Yan J, Li X, et al. (2010) Endocytic sorting and recycling require membrane phosphatidylserine asymmetry maintained by TAT-1/CHAT-1. PLoS Genet 6: e1001235. doi: 10.1371/journal.pgen.1001235. pmid:21170358
[32]
Woodman PG (2000) Biogenesis of the sorting endosome: the role of Rab5. Traffic 1: 695–701. pmid:11208157 doi: 10.1034/j.1600-0854.2000.010902.x
[33]
Ullrich O, Reinsch S, Urbe S, Zerial M, Parton RG (1996) Rab11 regulates recycling through the pericentriolar recycling endosome. J Cell Biol 135: 913–924. pmid:8922376 doi: 10.1083/jcb.135.4.913
[34]
Grosshans BL, Ortiz D, Novick P (2006) Rabs and their effectors: achieving specificity in membrane traffic. Proc Natl Acad Sci U S A 103: 11821–11827. pmid:16882731 doi: 10.1073/pnas.0601617103
[35]
Guo W, Roth D, Walch-Solimena C, Novick P (1999) The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis. EMBO J 18: 1071–1080. pmid:10022848 doi: 10.1093/emboj/18.4.1071
[36]
Jiu Y, Jin C, Liu Y, Holmberg CI, Jantti J (2012) Exocyst subunits Exo70 and Exo84 cooperate with small GTPases to regulate behavior and endocytic trafficking in C. elegans. PLoS One 7: e32077. doi: 10.1371/journal.pone.0032077. pmid:22389680
[37]
Armenti ST, Lohmer LL, Sherwood DR, Nance J (2014) Repurposing an endogenous degradation system for rapid and targeted depletion of C. elegans proteins. Development 141: 4640–4647. doi: 10.1242/dev.115048. pmid:25377555
[38]
Grindstaff KK, Yeaman C, Anandasabapathy N, Hsu SC, Rodriguez-Boulan E, et al. (1998) Sec6/8 complex is recruited to cell-cell contacts and specifies transport vesicle delivery to the basal-lateral membrane in epithelial cells. Cell 93: 731–740. pmid:9630218 doi: 10.1016/s0092-8674(00)81435-x
[39]
Tsuboi T, Ravier MA, Xie H, Ewart MA, Gould GW, et al. (2005) Mammalian exocyst complex is required for the docking step of insulin vesicle exocytosis. J Biol Chem 280: 25565–25570. pmid:15878854 doi: 10.1074/jbc.m501674200
[40]
Pereira-Leal JB, Seabra MC (2000) The mammalian Rab family of small GTPases: definition of family and subfamily sequence motifs suggests a mechanism for functional specificity in the Ras superfamily. J Mol Biol 301: 1077–1087. pmid:10966806 doi: 10.1006/jmbi.2000.4010
[41]
Shen Z, Zhang X, Chai Y, Zhu Z, Yi P, et al. (2014) Conditional knockouts generated by engineered CRISPR-Cas9 endonuclease reveal the roles of coronin in C. elegans neural development. Dev Cell 30: 625–636. doi: 10.1016/j.devcel.2014.07.017. pmid:25155554
[42]
Miyabayashi T, Palfreyman MT, Sluder AE, Slack F, Sengupta P (1999) Expression and function of members of a divergent nuclear receptor family in Caenorhabditis elegans. Dev Biol 215: 314–331. pmid:10545240 doi: 10.1006/dbio.1999.9470
[43]
Lerner DW, McCoy D, Isabella AJ, Mahowald AP, Gerlach GF, et al. (2013) A Rab10-dependent mechanism for polarized basement membrane secretion during organ morphogenesis. Dev Cell 24: 159–168. doi: 10.1016/j.devcel.2012.12.005. pmid:23369713
[44]
Jones TA, Nikolova LS, Schjelderup A, Metzstein MM (2014) Exocyst-mediated membrane trafficking is required for branch outgrowth in Drosophila tracheal terminal cells. Dev Biol 390: 41–50. doi: 10.1016/j.ydbio.2014.02.021. pmid:24607370
[45]
Chen S, Li L, Li J, Liu B, Zhu X, et al. (2014) SEC-10 and RAB-10 coordinate basolateral recycling of clathrin-independent cargo through endosomal tubules in Caenorhabditis elegans. Proc Natl Acad Sci U S A 111: 15432–15437. doi: 10.1073/pnas.1408327111. pmid:25301900
[46]
Jiu Y, Hasygar K, Tang L, Liu Y, Holmberg CI, et al. (2014) par-1, atypical pkc, and PP2A/B55 sur-6 are implicated in the regulation of exocyst-mediated membrane trafficking in Caenorhabditis elegans. G3 (Bethesda) 4: 173–183. doi: 10.1534/g3.113.006718
[47]
Murthy M, Teodoro RO, Miller TP, Schwarz TL (2010) Sec5, a member of the exocyst complex, mediates Drosophila embryo cellularization. Development 137: 2773–2783. doi: 10.1242/dev.048330. pmid:20630948
[48]
Einstein G, Buranosky R, Crain BJ (1994) Dendritic pathology of granule cells in Alzheimer's disease is unrelated to neuritic plaques. J Neurosci 14: 5077–5088. pmid:8046469
[49]
McNeill TH, Brown SA, Rafols JA, Shoulson I (1988) Atrophy of medium spiny I striatal dendrites in advanced Parkinson's disease. Brain Res 455: 148–152. pmid:3416180 doi: 10.1016/0006-8993(88)90124-2
[50]
Aguirre-Chen C, Bulow HE, Kaprielian Z (2011) C. elegans bicd-1, homolog of the Drosophila dynein accessory factor Bicaudal D, regulates the branching of PVD sensory neuron dendrites. Development 138: 507–518. doi: 10.1242/dev.060939. pmid:21205795
[51]
Rual JF, Ceron J, Koreth J, Hao T, Nicot AS, et al. (2004) Toward improving Caenorhabditis elegans phenome mapping with an ORFeome-based RNAi library. Genome Res 14: 2162–2168. pmid:15489339 doi: 10.1101/gr.2505604
[52]
Inoue T, Sherwood DR, Aspock G, Butler JA, Gupta BP, et al. (2002) Gene expression markers for Caenorhabditis elegans vulval cells. Mech Dev 119 Suppl 1: S203–209. pmid:14516686 doi: 10.1016/s0925-4773(03)00117-5
[53]
Gallegos ME, Balakrishnan S, Chandramouli P, Arora S, Azameera A, et al. (2012) The C. elegans rab family: identification, classification and toolkit construction. PLoS One 7: e49387. doi: 10.1371/journal.pone.0049387. pmid:23185324
[54]
Hobert O (2002) PCR fusion-based approach to create reporter gene constructs for expression analysis in transgenic C. elegans. Biotechniques 32: 728–730. pmid:11962590
[55]
Iyer EP, Iyer SC, Sullivan L, Wang D, Meduri R, et al. (2013) Functional genomic analyses of two morphologically distinct classes of Drosophila sensory neurons: post-mitotic roles of transcription factors in dendritic patterning. PLoS One 8: e72434. doi: 10.1371/journal.pone.0072434. pmid:23977298
