Plasticity-related gene 3 promotes neurite shaft protrusion

BMC Neuroscience

Anja U Bräuer¹  Ulf Strauss¹  Anna Soriguera Farrés¹  Beate Geist¹  Arne Battefeld¹  Tanja Velmans¹ 

[1] Institute of Cell Biology and Neurobiology, Center for Anatomy, Charité – Universitätsmedizin Berlin, Charitéplatz 1, Berlin, 10117, Germany

关键词: Growth cone;    Cytoskeleton;    Cell differentiation;    Phospholipids;   

Others  :  1140380
DOI  :  10.1186/1471-2202-14-36

 received in 2012-06-19, accepted in 2013-03-07,  发布年份 2013

【 摘 要 】

Background

Recently, we and others proposed plasticity-related gene 3 (PRG3) as a novel molecule in neuritogenesis based on PRG3 overexpression experiments in neuronal and non-neuronal cell lines. However, direct information on PRG3 effects in neuronal development and, in particular, its putative spatio-temporal distribution and conditions of action, is sparse.

Results

We demonstrate here that PRG3 induces filopodia formation in HEK293 cells depending on its N-glycosylation status. The PRG3 protein was strongly expressed during mouse brain development in vivo from embryonic day 16 to postnatal day 5 (E16 – P5). From P5 on, expression declined. Furthermore, in early, not yet polarized hippocampal cultured neurons, PRG3 was expressed along the neurite shaft. Knock-down of PRG3 in these neurons led to a decreased number of neurites. This phenotype is rescued by expression of an shRNA-resistant PRG3 construct in PRG3 knock-down neurons. After polarization, endogenous PRG3 expression shifted mainly to axons, specifically to the plasma membrane along the neurite shaft. These PRG3 pattern changes appeared temporally and spatially related to ongoing synaptogenesis. Therefore we tested (i) whether dendritic PRG3 re-enhancement influences synaptic currents and (ii) whether synaptic inputs contribute to the PRG3 shift. Our results rendered both scenarios unlikely: (i) PRG3 over-expression had no influence on miniature excitatory postsynaptic currents (mEPSC) and (ii) blocking of incoming signals did not alter PRG3 distribution dynamics. In addition, PRG3 levels did not interfere with intrinsic neuronal properties.

Conclusion

Taken together, our data indicate that endogenous PRG3 promotes neurite shaft protrusion and therefore contributes to regulating filopodia formation in immature neurons. PRG3 expression in more mature neurons, however, is predominantly localized in the axon. Changes in PRG3 levels did not influence intrinsic or synaptic neuronal properties.

【 授权许可】

   
2013 Velmans et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Song H, Poo M: The cell biology of neuronal navigation. Nat Cell Biol 2001, 3(3):E81-E88.
• [2]Luo L: Actin cytoskeleton regulation in neuronal morphogenesis and structural plasticity. Annu Rev Cell Dev Biol 2002, 18:601-635.
• [3]Dickson BJ: Molecular mechanisms of axon guidance. Science 2002, 298(5600):1959-1964.
• [4]Etienne-Manneville S: Cdc42–the centre of polarity. J Cell Sci 2004, 117(Pt 8):1291-1300.
• [5]Fukushima N, Furuta D, Hidaka Y, Moriyama R, Tsujiuchi T: Post-translational modifications of tubulin in the nervous system. J Neurochem 2009, 109(3):683-693.
• [6]Pan S, Iannotti MJ, Sifers RN: Analysis of serpin secretion, misfolding, and surveillance in the endoplasmic reticulum. Methods Enzymol 2011, 499:1-16.
• [7]Srikrishna G, Huttunen HJ, Johansson L, Weigle B, Yamaguchi Y, Rauvala H, Freeze HH: N -Glycans on the receptor for advanced glycation end products influence amphoterin binding and neurite outgrowth. J Neurochem 2002, 80(6):998-1008.
• [8]Dent EW, Gertler FB: Cytoskeletal dynamics and transport in growth cone motility and axon guidance. Neuron 2003, 40(2):209-227.
• [9]Schaefer AW, Kabir N, Forscher P: Filopodia and actin arcs guide the assembly and transport of two populations of microtubules with unique dynamic parameters in neuronal growth cones. J Cell Biol 2002, 158(1):139-152.
• [10]Mingorance-Le Meur A, O'Connor TP: Neurite consolidation is an active process requiring constant repression of protrusive activity. EMBO J 2009, 28(3):248-260.
• [11]Welch MD, Mullins RD: Cellular control of actin nucleation. Annu Rev Cell Dev Biol 2002, 18:247-288.
• [12]Fukushima N, Weiner JA, Chun J: Lysophosphatidic acid (LPA) is a novel extracellular regulator of cortical neuroblast morphology. Dev Biol 2000, 228(1):6-18.
• [13]Fukushima N, Morita Y: Actomyosin-dependent microtubule rearrangement in lysophosphatidic acid-induced neurite remodeling of young cortical neurons. Brain Res 2006, 1094(1):65-75.
• [14]Moolenaar WH: Lysophosphatidic acid, a multifunctional phospholipid messenger. J Biol Chem 1995, 270(22):12949-12952.
• [15]Moolenaar WH, Kranenburg O, Postma FR, Zondag GC: Lysophosphatidic acid: G-protein signalling and cellular responses. Curr Opin Cell Biol 1997, 9(2):168-173.
• [16]Brindley DN: Lipid phosphate phosphatases and related proteins: signaling functions in development, cell division, and cancer. J Cell Biochem 2004, 92(5):900-912.
• [17]Brauer AU, Savaskan NE, Kuhn H, Prehn S, Ninnemann O, Nitsch R: A new phospholipid phosphatase, PRG-1, is involved in axon growth and regenerative sprouting. Nat Neurosci 2003, 6(6):572-578.
• [18]Savaskan NE, Brauer AU, Nitsch R: Molecular cloning and expression regulation of PRG-3, a new member of the plasticity-related gene family. Eur J Neurosci 2004, 19(1):212-220.
• [19]Wang WZ, Molnar Z: Dynamic pattern of mRNA expression of plasticity-related gene-3 (PRG-3) in the mouse cerebral cortex during development. Brain Res Bull 2005, 66(4–6):454-460.
• [20]Sigal YJ, Quintero OA, Cheney RE, Morris AJ: Cdc42 and ARP2/3-independent regulation of filopodia by an integral membrane lipid-phosphatase-related protein. J Cell Sci 2007, 120(Pt 2):340-352.
• [21]Savaskan NE, Rocha L, Kotter MR, Baer A, Lubec G, van Meeteren LA, Kishi Y, Aoki J, Moolenaar WH, Nitsch R: Autotaxin (NPP-2) in the brain: cell type-specific expression and regulation during development and after neurotrauma. Cell Mol Life Sci 2007, 64(2):230-243.
• [22]Drews D, Klar M, Dame C, Brauer AU: Developmental expression profile of the YY2 gene in mice. BMC Dev Biol 2009, 9:45. BioMed Central Full Text
• [23]Elder JH, Alexander S: endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci USA 1982, 79(15):4540-4544.
• [24]Cabral CM, Liu Y, Sifers RN: Dissecting glycoprotein quality control in the secretory pathway. Trends Biochem Sci 2001, 26(10):619-624.
• [25]Zhang QX, Pilquil CS, Dewald J, Berthiaume LG, Brindley DN: Identification of structurally important domains of lipid phosphate phosphatase-1: implications for its sites of action. Biochem J 2000, 345(Pt 2):181-184.
• [26]Wallace DF, Harris JM, Subramaniam VN: Functional analysis and theoretical modeling of ferroportin reveals clustering of mutations according to phenotype. Am J Physiol Cell Physiol 2009, 298(1):C75-C84.
• [27]Grabrucker A, Vaida B, Bockmann J, Boeckers TM: Synaptogenesis of hippocampal neurons in primary cell culture. Cell Tissue Res 2009, 338(3):333-341.
• [28]Barila D, Plateroti M, Nobili F, Muda AO, Xie Y, Morimoto T, Perozzi G: The Dri 42 gene, whose expression is up-regulated during epithelial differentiation, encodes a novel endoplasmic reticulum resident transmembrane protein. J Biol Chem 1996, 271(47):29928-29936.
• [29]Trimbuch T, Beed P, Vogt J, Schuchmann S, Maier N, Kintscher M, Breustedt J, Schuelke M, Streu N, Kieselmann O: Synaptic PRG-1 modulates excitatory transmission via lipid phosphate-mediated signaling. Cell 2009, 138(6):1222-1235.
• [30]Noguchi K, Herr D, Mutoh T, Chun J: Lysophosphatidic acid (LPA) and its receptors. Curr Opin Pharmacol 2009, 9(1):15-23.
• [31]van Horck FP, Weinl C, Holt CE: Retinal axon guidance: novel mechanisms for steering. Curr Opin Neurobiol 2004, 14(1):61-66.
• [32]Broggini T, Nitsch R, Savaskan NE: Plasticity-related gene 5 (PRG5) induces filopodia and neurite growth and impedes lysophosphatidic acid- and nogo-A-mediated axonal retraction. Mol Biol Cell 2010, 21(4):521-537.
• [33]Wiggin GR, Fawcett JP, Pawson T: Polarity proteins in axon specification and synaptogenesis. Dev Cell 2005, 8(6):803-816.
• [34]Gomperts SN, Carroll R, Malenka RC, Nicoll RA: Distinct roles for ionotropic and metabotropic glutamate receptors in the maturation of excitatory synapses. J Neurosci 2000, 20(6):2229-2237.
• [35]Basarsky TA, Parpura V, Haydon PG: Hippocampal synaptogenesis in cell culture: developmental time course of synapse formation, calcium influx, and synaptic protein distribution. J Neurosci 1994, 14(11 Pt 1):6402-6411.