Collapsin response mediator protein 5 accelerates neurite outgrowth in hippocampal neurons

doi:10.3969/j.issn.0529-1356.2014.03.001

Abstract

Abstract:

Objective To investigate function of collapsin response mediator protein 5(CRMP5) on neurite outgrowth. Methods The CRMP5 eukaryotic expression vector was constructed and transfected into hippocampal neurons. The gene transfection, Real-time quantitative PCR and Western blotting were used to detect expression of CRMP5 protein. The lapse-time imaging and neurite extraction were utilized to show neurite outgrowth and differentiation and 3 double-pored were performed, compared with the vector without CRMP5 gene. Results It was successful to construct the CRMP5 eukaryotic expression vector with an EGFP tag. The lipofectamine effectually transfected CRMP5 into cultured neurons, and the CRMP5 protein was expressed successfully more than the control cells. CRMP5 protein was abundant in cell body, initiation and end of neurites. Overexpression of CRMP5 in neuronal cells significantly promoted outgrowth neurites, and led to the formation of longer neurites with more branches. Accompanying rapid outgrowth of neurites, branches from original neurites were contributed to form a network. The results of neurite length and extraction showed that neurons overexpressing CRMP5 were possessed more and longer neurites (P<0.01), compared with control cells. Conclusion The results suggest that CRMP5 accelerates not only axonal growth but also branching.

Key words: Collapsin response mediator protein 5, Neuron, Neurite, Outgrowth, Gene transfection, Eukaryotic expression, Western blotting, Rat

CHEN Ke-en WANG Yuan-yuan ZHANG Ji-feng LI Bin GUO Guo-qing*. Collapsin response mediator protein 5 accelerates neurite outgrowth in hippocampal neurons[J]. AAS, 2014, 45(3): 297-303.

References

［1］Bretin S, Reibel S, Charrier E, et al. Differential expression of CRMP1, CRMP2A, CRMP2B, and CRMP5 in axons or dendrites of distinct neurons in the mouse brain［J］. J Comp Neurol, 2005, 486(1):1-17.
［2］Arimura N, Kaibuchi K. Neuronal polarity: from extracellular signals to intracellular mechanisms［J］. Nat Rev Neurosci, 2007, 8(3):194-205.
［3］Horiuchi M, El Far O, Betz H. Ulip6, a novel unc-33 and dihydropyrimidinase related protein highly expressed in developing rat brain［J］. FEBS Lett, 2000, 480(2-3):283-286.
［4］Fukada M, Watakabe I, Yuasa-Kawada J, et al. Molecular characterization of CRMP5, a novel member of the collapsin response mediator protein family［J］. J Biol Chem, 2000, 275(48): 37957-37965.
［5］Veyrac A, Reibel S, Sacquet J, et al. CRMP5 regulates generation and survival of newborn neurons in olfactory and hippocampal neurogenic areas of the adult mouse brain［J］. PLoS One, 2011, 6(10): e23721.
［6］Cross SA, Salomao DR, Parisi JE, et al. Paraneoplastic autoimmune optic neuritis with retinitis defined by CRMP-5-IgG［J］. Ann Neurol, 2003, 54(1):38-50.
［7］Auvergnon N, Reibel S, Touret, M, et al. Altered expression of CRMPs in the brain of bovine spongiform encephalopathy-infected mice during disease progression［J］. Brain Res, 2009, 1261:1-6.
［8］Byk T, Haddada H, Vainchenker W, et al. Lipofectamine and related cationic lipids strongly improve adenoviral infection efficiency of primitive human hematopoietic cells［J］. Hum Gene Ther, 1998, 9(17):2493-2502.
［9］Kong L，Zhou GH，Yang GJ，et al. Construction of CTSB-RNAi-LV vector and its effect on mouse brain micvascular endothelial cell line bEND.3［J］. Acta Anatomica Sinica, 2011, 42(2):190-194.（in Chinese）
孔丽，周国宏，杨桂姣，等. 组织蛋白酶BRNAiLV载体构建及其对小鼠脑微血管内皮细胞系bEND.3的影响［J］. 解剖学报，2011, 42(2):190-194. 
［10］Cnops L, Van de Plas B, Arckens L. Age-dependent expression of collapsin response mediator proteins (CRMPs) in cat visual cortex［J］. Eur J Neurosci, 2004, 19(8):2345-2351.
［11］Cnops L, Hu TT, Burnat K, et al. Age-dependent alterations in CRMP2 and CRMP4 protein expression profiles in cat visual cortex［J］. Brain Res, 2006, 1088(1):109-119.
［12］Fukada M, Watakabe I, Yuasa-Kawada J, et al. Molecular characterization of CRMP5, a novel member of the collapsin response mediator protein family［J］. J Biol Chem, 2000, 275(48): 37957-37965.
［13］Rasband MN. The axon initial segment and the maintenance of neuronal polarity［J］. Nat Rev Neurosci, 2010, 11(8):552-562.
［14］SChmidt H, Rathjen FG. Signalling mechanisms regulating axonal branching in vivo［J］. Bioessays, 2010, 32(11):977-985.
［15］Lin PC, Chan PM, Hall C, et al. Collapsin response mediator proteins (CRMPs) are a new class of microtubule-associated protein (MAP) that selectively interacts with assembled microtubules via a taxol-sensitive binding interaction［J］. J Biol Chem, 2011, 286(48): 41466-41478.
［16］Fukata Y, Itoh TJ, Kimura T, et al. CRMP-2 binds to tubulin heterodimers to promote microtubule assembly［J］. Nat Cell Biol, 2002, 4(8):583-591.
［17］Yoshimura T, Kawano Y, Arimura N, et al. GSK-3beta regulates phosphorylation of CRMP-2 and neuronal polarity［J］. Cell, 2005, 120(1):137-149.
［18］Brot S, Rogemond V, Perrot V, et al. CRMP5 interacts with tubulin to inhibit neurite outgrowth,thereby modulating the function of CRMP2［J］.J Neurosci, 2010, 30(32):10639-10654
［19］Yamashita N, Mosinger B, Roy A, et al. CRMP5 (collapsin response mediator protein 5) regulates dendritic development and synaptic plasticity in the cerebellar Purkinje cells［J］. J Neurosci, 2011, 31(5):1773-1779.
［20］Hotta A, Inatome R, Yuasa-Kawada J, et al. Critical role of collapsin response mediator protein associated molecule CRAM for filopodia and growth cone development in neurons［J］. Mol Biol Cell, 2005, 16(1): 32-39.

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