Abstract
Recent studies have demonstrated an important role for miR-124, the most abundant and well-conserved brain-specific microRNA(miRNA), in promoting neurite outgrowth and elongation during neuronal differentiation. This miRNA’s target genes and the mechanisms that execute this role remain unclear. In this study, we identified ROCK1, a small GTPase Rho kinase, as a direct target of miR-124 for regulating neurite elongation. miR-124 significantly inhibited ROCK1 expression in M17 cells. Inhibiting ROCK1 promoted neurite elongation, and the overexpression of ROCK1 strongly repressed the neurite elongation-enhancing effect of miR-124 in M17 cells. We determined that Akt functions as a novel ROCK1 downstream effector in regulating neurite outgrowth and elongation.
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Arimura N, Inagaki N, Chihara K, Menager C, Nakamura N, Amano M, Iwamatsu A, Goshima Y, Kaibuchi K (2000) Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. evidence for two separate signaling pathways for growth cone collapse. J Biological Chemistry 275:23973–23980
Arimura N, Menager C, Kawano Y, Yoshimura T, Kawabata S, Hattori A, Fukata Y, Amano M, Goshima Y, Inagaki M, Morone N, Usukura J, Kaibuchi K (2005) Phosphorylation by Rho kinase regulates CRMP-2 activity in growth cones. Mol Cell Biol 25:9973–9984
Baudet ML, Zivraj KH, Abreu-Goodger C, Muldal A, Armisen J, Blenkiron C, Goldstein LD, Miska EA, Holt CE (2012) miR-124 acts through CoREST to control onset of Sema3A sensitivity in navigating retinal growth cones. Nat Neurosci 15:29–38
Brandt N, Franke K, Rasin MR, Baumgart J, Vogt J, Khrulev S, Hassel B, Pohl EE, Sestan N, Nitsch R, Schumacher S (2007) The neural EGF family member CALEB/NGC mediates dendritic tree and spine complexity. EMBO J 26:2371–2386
Cheng LC, Pastrana E, Tavazoie M, Doetsch F (2009) miR-124 regulates adult neurogenesis in the subventricular zone stem cell niche. Nat Neurosci 12:399–408
Chiang HR, Schoenfeld LW, Ruby JG, Auyeung VC, Spies N, Baek D, Johnston WK, Russ C, Luo S, Babiarz JE, Blelloch R, Schroth GP, Nusbaum C, Bartel DP (2010) Mammalian microRNAs: experimental evaluation of novel and previously annotated genes. Genes Development 24:992–1009
Da Silva JS, Medina M, Zuliani C, Di Nardo A, Witke W, Dotti CG (2003) RhoA/ROCK regulation of neuritogenesis via profilin IIa-mediated control of actin stability. J Cell Biology 162:1267–1279
Del Rio JA, Gonzalez-Billault C, Urena JM, Jimenez EM, Barallobre MJ, Pascual M, Pujadas L, Simo S, La Torre A, Wandosell F, Avila J, Soriano E (2004) MAP1B is required for Netrin 1 signaling in neuronal migration and axonal guidance. Current Biology: CB 14:840–850
Dertinger SK, Jiang X, Li Z, Murthy VN, Whitesides GM (2002) Gradients of substrate-bound laminin orient axonal specification of neurons. Proc Natl Acad Sci U S A 99:12542–12547
Duffy P, Schmandke A, Schmandke A, Sigworth J, Narumiya S, Cafferty WB, Strittmatter SM (2009) Rho-associated kinase II (ROCKII) limits axonal growth after trauma within the adult mouse spinal cord. J Neurosci: Off J Soc Neurosci 29:15266–15276
Flynt AS, Lai EC (2008) Biological principles of microRNA-mediated regulation: shared themes amid diversity. Nat Rev Genet 9:831–842
Franke K, Otto W, Johannes S, Baumgart J, Nitsch R, Schumacher S (2012) miR-124-regulated RhoG reduces neuronal process complexity via ELMO/Dock180/Rac1 and Cdc42 signalling. EMBO J 31:2908–2921
Fujisawa K, Fujita A, Ishizaki T, Saito Y, Narumiya S (1996) Identification of the Rho-binding domain of p160ROCK, a Rho-associated coiled-coil containing protein kinase. J Biological Chemistry 271:23022–23028
Gomes C, Smith SC, Youssef MN, Zheng JJ, Hagg T, Hetman M (2011) RNA polymerase 1-driven transcription as a mediator of BDNF-induced neurite outgrowth. J Biological Chemistry 286:4357–4363
Govek EE, Newey SE, Van Aelst L (2005) The role of the Rho GTPases in neuronal development. Genes Development 19:1–49
Hirose M, Ishizaki T, Watanabe N, Uehata M, Kranenburg O, Moolenaar WH, Matsumura F, Maekawa M, Bito H, Narumiya S (1998) Molecular dissection of the Rho-associated protein kinase (p160ROCK)-regulated neurite remodeling in neuroblastoma N1E-115 cells. J Cell Biology 141:1625–1636
Huang B, Li W, Zhao B, **a C, Liang R, Ruan K, **g N, ** Y (2009) MicroRNA expression profiling during neural differentiation of mouse embryonic carcinoma P19 cells. Acta Biochim Biophys Sin 41:231–236
Huang TC, Chang HY, Chen CY, Wu PY, Lee H, Liao YF, Hsu WM, Huang HC, Juan HF (2011) Silencing of miR-124 induces neuroblastoma SK-N-SH cell differentiation, cell cycle arrest and apoptosis through promoting AHR. FEBS Lett 585:3582–3586
Jaworski J, Spangler S, Seeburg DP, Hoogenraad CC, Sheng M (2005) Control of dendritic arborization by the phosphoinositide-3′-kinase-Akt-mammalian target of rapamycin pathway. J Neurosci: Off J Soc Neurosci 25:11300–11312
Krichevsky AM, Sonntag KC, Isacson O, Kosik KS (2006) Specific microRNAs modulate embryonic stem cell-derived neurogenesis. Stem Cells 24:857–864
Lang Q, Ling C (2012) MiR-124 suppresses cell proliferation in hepatocellular carcinoma by targeting PIK3CA. Biochem Biophys Res Commun 426:247–252
Lee JH, Shin SY, Kim S, Choo J, Lee YH (2006) Suppression of PTEN expression during aggregation with retinoic acid in P19 mouse embryonal carcinoma cells. Biochem Biophys Res Comm 347:715–722
Luo L (2000) Rho GTPases in neuronal morphogenesis. Nat Rev Neurosci 1:173–180
Maekawa M, Ishizaki T, Boku S, Watanabe N, Fujita A, Iwamatsu A, Obinata T, Ohashi K, Mizuno K, Narumiya S (1999) Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science 285:895–898
Makeyev EV, Zhang J, Carrasco MA, Maniatis T (2007) The microRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol Cell 27:435–448
Mueller BK, Mack H, Teusch N (2005) Rho kinase, a promising drug target for neurological disorders. Nat Rev Drug Discov 4:387–398
Nakayama M, Goto TM, Sugimoto M, Nishimura T, Shinagawa T, Ohno S, Amano M, Kaibuchi K (2008) Rho-kinase phosphorylates PAR-3 and disrupts PAR complex formation. Dev Cell 14:205–215
Shi SH, Jan LY, Jan YN (2003) Hippocampal neuronal polarity specified by spatially localized mPar3/mPar6 and PI 3-kinase activity. Cell 112:63–75
Sklan EH, Berson A, Birikh KR, Gutnick A, Shahar O, Shoham S, Soreq H (2006) Acetylcholinesterase modulates stress-induced motor responses through catalytic and noncatalytic properties. Biol Psychiatry 60:741–751
Stefani G, Slack FJ (2008) Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol 9:219–230
Visvanathan J, Lee S, Lee B, Lee JW, Lee SK (2007) The microRNA miR-124 antagonizes the anti-neural REST/SCP1 pathway during embryonic CNS development. Genes Development 21:744–749
Yu JY, Chung KH, Deo M, Thompson RC, Turner DL (2008) MicroRNA miR-124 regulates neurite outgrowth during neuronal differentiation. Exp Cell Res 314:2618–2633
Zhang X, Zhu J, Yang GY, Wang QJ, Qian L, Chen YM, Chen F, Tao Y, Hu HS, Wang T, Luo ZG (2007) Dishevelled promotes axon differentiation by regulating atypical protein kinase C. Nature cell Biology 9:743–754
Zheng F, Liao YJ, Cai MY, Liu YH, Liu TH, Chen SP, Bian XW, Guan XY, Lin MC, Zeng YX, Kung HF, **e D (2012) The putative tumour suppressor microRNA-124 modulates hepatocellular carcinoma cell aggressiveness by repressing ROCK2 and EZH2. Gut 61:278–289
Acknowledgements
This work was supported by the supported by the National Natural Science Foundation of China (81071016), Program for New Century Excellent Talents in University (NCET-12-0647), Key Project of Guangdong Province (CXZD1018), and the Program for Changjiang Scholars and the Innovative Research Team in University (IRT1142). The authors declare that they have no conflicts of interest.
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Gu, X., Meng, S., Liu, S. et al. miR-124 Represses ROCK1 Expression to Promote Neurite Elongation Through Activation of the PI3K/Akt Signal Pathway. J Mol Neurosci 52, 156–165 (2014). https://doi.org/10.1007/s12031-013-0190-6
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DOI: https://doi.org/10.1007/s12031-013-0190-6