Abstract
Nogo-A is a well-known myelin-enriched inhibitory protein for axonal growth and regeneration in the central nervous system (CNS). Besides oligodendrocytes, our previous data revealed that Nogo-A is also expressed in subpopulations of neurons including retinal ganglion cells, in which it can have a positive role in the neuronal growth response after injury, through an unclear mechanism. In the present study, we analyzed the opposite roles of glial versus neuronal Nogo-A in the injured visual system. To this aim, we created oligodendrocyte (Cnp-Cre+/−xRtn4/Nogo-Aflox/flox) and neuron-specific (Thy1-Cretg+xRtn4flox/flox) conditional Nogo-A knock-out (KO) mouse lines. Following complete intraorbital optic nerve crush, both spontaneous and inflammation-mediated axonal outgrowth was increased in the optic nerves of the glia-specific Nogo-A KO mice. In contrast, neuron-specific deletion of Nogo-A in a KO mouse line or after acute gene recombination in retinal ganglion cells mediated by adeno-associated virus serotype 2.Cre virus injection in Rtn4flox/flox animals decreased axon sprouting in the injured optic nerve. These results therefore show that selective ablation of Nogo-A in oligodendrocytes and myelin in the optic nerve is more effective at enhancing regrowth of injured axons than what has previously been observed in conventional, complete Nogo-A KO mice. Our data also suggest that neuronal Nogo-A in retinal ganglion cells could participate in enhancing axonal sprouting, possibly by cis-interaction with Nogo receptors at the cell membrane that may counteract trans-Nogo-A signaling. We propose that inactivating Nogo-A in glia while preserving neuronal Nogo-A expression may be a successful strategy to promote axonal regeneration in the CNS.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- AAV:
-
adeno-associated virus
- AKT:
-
protein kinase B
- APC:
-
adenomatous polyposis coli
- CNPase:
-
2′,3′-cyclic nucleotide 3′-phosphodiesterase
- CNS:
-
central nervous system
- CRALBP:
-
cellular retinaldehyde binding protein
- CREB:
-
cAMP response element-binding protein
- CTB-A594:
-
cholera toxin B-Alexa594
- DRG:
-
dorsal root ganglion
- ERK1/2:
-
extracellular signal-regulated kinases 1/2
- Gap-43:
-
growth-associated protein 43
- GAPDH:
-
glyceraldehyde 3-phosphate dehydrogenase
- GFP:
-
green fluorescent protein
- KO:
-
knockout
- MAG:
-
myelin-associated glycoprotein
- NgR1:
-
Nogo66 receptor 1
- ONC:
-
optic nerve crush injury
- PBS:
-
phosphate-buffered saline
- qRT-PCR:
-
quantitative real-time polymerase chain reaction
- RGC:
-
retinal ganglion cell
- RTN:
-
reticulon
- SOCS3:
-
suppressor of cytokine signaling 3
- S1PR2:
-
sphingosine 1-phosphate receptor 2
- Sprr1A:
-
small proline-rich protein 1A
- STAT3:
-
signal transducer and activator of transcription 3
- TBS:
-
Tris-buffered saline
References
Yiu G, He Z . Glial inhibition of CNS axon regeneration. Nat Rev Neurosci 2006; 7: 617–627.
Schwab ME . Functions of Nogo proteins and their receptors in the nervous system. Nat Rev Neurosci 2010; 11: 799–811.
Goldberg JL, Espinosa JS, Xu Y, Davidson N, Kovacs GT, Barres BA . Retinal ganglion cells do not extend axons by default: promotion by neurotrophic signaling and electrical activity. Neuron 2002; 33: 689–702.
Schwab ME . Nogo and axon regeneration. Curr Opin Neurobiol 2004; 14: 118–124.
Pernet V, Schwab ME . The role of Nogo-A in axonal plasticity, regrowth and repair. Cell Tissue Res 2012; 349: 97–104.
Bareyre FM, Haudenschild B, Schwab ME . Long-lasting sprouting and gene expression changes induced by the monoclonal antibody IN-1 in the adult spinal cord. J Neurosci 2002; 22: 7097–7110.
Simonen M, Pedersen V, Weinmann O, Schnell L, Buss A, Ledermann B et al. Systemic deletion of the myelin-associated outgrowth inhibitor Nogo-A improves regenerative and plastic responses after spinal cord injury. Neuron 2003; 38: 201–211.
Thallmair M, Metz GA, Z'Graggen WJ, Raineteau O, Kartje GL, Schwab ME . Neurite growth inhibitors restrict plasticity and functional recovery following corticospinal tract lesions. Nat Neurosci 1998; 1: 124–131.
Dimou L, Schnell L, Montani L, Duncan C, Simonen M, Schneider R et al. Nogo-A-deficient mice reveal strain-dependent differences in axonal regeneration. J Neurosci 2006; 26: 5591–5603.
Kim JE, Li S, GrandPre T, Qiu D, Strittmatter SM . Axon regeneration in young adult mice lacking Nogo-A/B. Neuron 2003; 38: 187–199.
Schwab ME, Strittmatter SM . Nogo limits neural plasticity and recovery from injury. Curr Opin Neurobiol 2014; 27C: 53–60.
Huber AB, Weinmann O, Brosamle C, Oertle T, Schwab ME . Patterns of Nogo mRNA and protein expression in the developing and adult rat and after CNS lesions. J Neurosci 2002; 22: 3553–3567.
Wang X, Chun SJ, Treloar H, Vartanian T, Greer CA, Strittmatter SM . Localization of Nogo-A and Nogo-66 receptor proteins at sites of axon-myelin and synaptic contact. J Neurosci 2002; 22: 5505–5515.
Fournier AE, GrandPre T, Strittmatter SM . Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration. Nature 2001; 409: 341–346.
Kempf A, Tews B, Arzt ME, Weinmann O, Obermair FJ, Pernet V et al. The sphingolipid receptor S1PR2 is a receptor for Nogo-a repressing synaptic plasticity. PLoS Biol 2014; 12: e1001763.
Egea J, Klein R . Bidirectional Eph-ephrin signaling during axon guidance. Trends Cell Biol 2007; 17: 230–238.
Haklai-Topper L, Mlechkovich G, Savariego D, Gokhman I, Yaron A . Cis interaction between Semaphorin6A and Plexin-A4 modulates the repulsive response to Sema6A. EMBO J 2010; 29: 2635–2645.
Delekate A, Zagrebelsky M, Kramer S, Schwab ME, Korte M . NogoA restricts synaptic plasticity in the adult hippocampus on a fast time scale. Proc Natl Acad Sci USA 2011; 108: 2569–2574.
Tews B, Schonig K, Arzt ME, Clementi S, Rioult-Pedotti MS, Zemmar A et al. Synthetic microRNA-mediated downregulation of Nogo-A in transgenic rats reveals its role as regulator of synaptic plasticity and cognitive function. Proc Natl Acad Sci USA 2013; 110: 6583–6588.
Wills ZP, Mandel-Brehm C, Mardinly AR, McCord AE, Giger RJ, Greenberg ME . The nogo receptor family restricts synapse number in the developing hippocampus. Neuron 2012; 73: 466–481.
Raiker SJ, Lee H, Baldwin KT, Duan Y, Shrager P, Giger RJ . Oligodendrocyte-myelin glycoprotein and Nogo negatively regulate activity-dependent synaptic plasticity. J Neurosci 2010; 30: 12432–12445.
Akbik FV, Bhagat SM, Patel PR, Cafferty WB, Strittmatter SM . Anatomical plasticity of adult brain is titrated by Nogo Receptor 1. Neuron 2013; 77: 859–866.
Mathis C, Schroter A, Thallmair M, Schwab ME . Nogo-a regulates neural precursor migration in the embryonic mouse cortex. Cereb Cortex 2010; 20: 2380–2390.
Mingorance-Le Meur A, Zheng B, Soriano E, del Rio JA . Involvement of the myelin-associated inhibitor Nogo-A in early cortical development and neuronal maturation. Cereb Cortex 2007; 17: 2375–2386.
Mi YJ, Hou B, Liao QM, Ma Y, Luo Q, Dai YK et al. Amino-Nogo-A antagonizes reactive oxygen species generation and protects immature primary cortical neurons from oxidative toxicity. Cell Death Differ 2012; 19: 1175–1186.
Guo F, Jin WL, Li LY, Song WY, Wang HW, Gou XC et al. M9, a novel region of amino-Nogo-A, attenuates cerebral ischemic injury by inhibiting NADPH oxidase-derived superoxide production in mice. CNS Neurosci Ther 2013; 19: 319–328.
Oertle T, van der Haar ME, Bandtlow CE, Robeva A, Burfeind P, Buss A et al. Nogo-A inhibits neurite outgrowth and cell spreading with three discrete regions. J Neurosci 2003; 23: 5393–5406.
Petrinovic MM, Duncan CS, Bourikas D, Weinman O, Montani L, Schroeter A et al. Neuronal Nogo-A regulates neurite fasciculation, branching and extension in the developing nervous system. Development 2010; 137: 2539–2550.
Pernet V, Joly S, Dalkara D, Schwarz O, Christ F, Schaffer D et al. Neuronal Nogo-A upregulation does not contribute to ER stress-associated apoptosis but participates in the regenerative response in the axotomized adult retina. Cell Death Differ 2011; 19: 1096–1108.
Lappe-Siefke C, Goebbels S, Gravel M, Nicksch E, Lee J, Braun PE et al. Disruption of Cnp1 uncouples oligodendroglial functions in axonal support and myelination. Nat Genet 2003; 33: 366–374.
Sprinkle TJ . 2',3'-cyclic nucleotide 3'-phosphodiesterase, an oligodendrocyte-Schwann cell and myelin-associated enzyme of the nervous system. Crit Rev Neurobiol 1989; 4: 235–301.
Aigner L, Arber S, Kapfhammer JP, Laux T, Schneider C, Botteri F et al. Overexpression of the neural growth-associated protein GAP-43 induces nerve sprouting in the adult nervous system of transgenic mice. Cell 1995; 83: 269–278.
Bonilla IE, Tanabe K, Strittmatter SM . Small proline-rich repeat protein 1A is expressed by axotomized neurons and promotes axonal outgrowth. J Neurosci 2002; 22: 1303–1315.
Pernet V, Joly S, Jordi N, Dalkara D, Guzik-Kornacka A, Flannery JG et al. Misguidance and modulation of axonal regeneration by Stat3 and Rho/ROCK signaling in the transparent optic nerve. Cell Death Dis 2013; 4: e734.
Yin Y, Cui Q, Gilbert HY, Yang Y, Yang Z, Berlinicke C et al. Oncomodulin links inflammation to optic nerve regeneration. Proc Natl Acad Sci USA 2009; 106: 19587–19592.
Yin Y, Henzl MT, Lorber B, Nakazawa T, Thomas TT, Jiang F et al. Oncomodulin is a macrophage-derived signal for axon regeneration in retinal ganglion cells. Nat Neurosci 2006; 9: 843–852.
Luo X, Salgueiro Y, Beckerman SR, Lemmon VP, Tsoulfas P, Park KK . Three-dimensional evaluation of retinal ganglion cell axon regeneration and pathfinding in whole mouse tissue after injury. Exp Neurol 2013; 247: 653–662.
Dodt HU, Leischner U, Schierloh A, Jahrling N, Mauch CP, Deininger K et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat Methods 2007; 4: 331–336.
Kempf A, Montani L, Petrinovic MM, Schroeter A, Weinmann O, Patrignani A et al. Upregulation of axon guidance molecules in the adult central nervous system of Nogo-A knockout mice restricts neuronal growth and regeneration. Eur J Neurosci 2013; 38: 3567–3579.
Joly S, Jordi N, Schwab ME, Pernet V . The Ephrin receptor EphA4 restricts axonal sprouting and enhances branching in the injured mouse optic nerve. Eur J Neurosci 2014 e-pub ahead of print 12 July 2014 doi:10.1111/ejn.12677.
Cafferty WB, Duffy P, Huebner E, Strittmatter SM . MAG and OMgp synergize with Nogo-A to restrict axonal growth and neurological recovery after spinal cord trauma. J Neurosci 2010; 30: 6825–6837.
Moore DL, Blackmore MG, Hu Y, Kaestner KH, Bixby JL, Lemmon VP et al. KLF family members regulate intrinsic axon regeneration ability. Science 2009; 326: 298–301.
Liebscher T, Schnell L, Schnell D, Scholl J, Schneider R, Gullo M et al. Nogo-A antibody improves regeneration and locomotion of spinal cord-injured rats. Ann Neurol 2005; 58: 706–719.
Li S, Liu BP, Budel S, Li M, Ji B, Walus L et al. Blockade of Nogo-66, myelin-associated glycoprotein, and oligodendrocyte myelin glycoprotein by soluble Nogo-66 receptor promotes axonal sprouting and recovery after spinal injury. J Neurosci 2004; 24: 10511–10520.
Dergham P, Ellezam B, Essagian C, Avedissian H, Lubell WD, McKerracher L . Rho signaling pathway targeted to promote spinal cord repair. J Neurosci 2002; 22: 6570–6577.
Sun F, Park KK, Belin S, Wang D, Lu T, Chen G et al. Sustained axon regeneration induced by co-deletion of PTEN and SOCS3. Nature 2011; 480: 372–375.
Yin Y, Cui Q, Li Y, Irwin N, Fischer D, Harvey AR et al. Macrophage-derived factors stimulate optic nerve regeneration. J Neurosci 2003; 23: 2284–2293.
Hauk TG, Leibinger M, Muller A, Andreadaki A, Knippschild U, Fischer D . Stimulation of axon regeneration in the mature optic nerve by intravitreal application of the toll-like receptor 2 agonist Pam3Cys. Invest Ophthalmol Vis Sci 2010; 51: 459–464.
Wang X, Hasan O, Arzeno A, Benowitz LI, Cafferty WB, Strittmatter SM . Axonal regeneration induced by blockade of glial inhibitors coupled with activation of intrinsic neuronal growth pathways. Exp Neurol 2012; 237: 55–69.
Fischer D, He Z, Benowitz LI . Counteracting the Nogo receptor enhances optic nerve regeneration if retinal ganglion cells are in an active growth state. J Neurosci 2004; 24: 1646–1651.
Dickendesher TL, Baldwin KT, Mironova YA, Koriyama Y, Raiker SJ, Askew KL et al. NgR1 and NgR3 are receptors for chondroitin sulfate proteoglycans. Nat Neurosci 2012; 15: 703–712.
Giger RJ, Hollis 2nd ER, Tuszynski MH . Guidance molecules in axon regeneration. Cold Spring Harb Perspect Biol 2010; 2: a001867.
Pernet V, Schwab ME . Lost in the jungle: new hurdles for optic nerve axon regeneration. Trends Neurosci 2014; 37: 381–387.
Kern F, Stanika RI, Sarg B, Offterdinger M, Hess D, Obermair GJ et al. Nogo-A couples with Apg-1 through interaction and co-ordinate expression under hypoxic and oxidative stress. Biochem J 2013; 455: 217–227.
Dubreuil CI, Winton MJ, McKerracher L . Rho activation patterns after spinal cord injury and the role of activated Rho in apoptosis in the central nervous system. J Cell Biol 2003; 162: 233–243.
Jaworski A, Tessier-Lavigne M . Autocrine/juxtaparacrine regulation of axon fasciculation by Slit-Robo signaling. Nat Neurosci 2012; 15: 367–369.
Sprinzak D, Lakhanpal A, Lebon L, Santat LA, Fontes ME, Anderson GA et al. Cis-interactions between Notch and Delta generate mutually exclusive signalling states. Nature 2010; 465: 86–90.
Emmenlauer M, Ronneberger O, Ponti A, Schwarb P, Griffa A, Filippi A et al. XuvTools: free, fast and reliable stitching of large 3D datasets. J Microsc 2009; 233: 42–60.
Erb M, Steck AJ, Nave KA, Schaeren-Wiemers N . Differential expression of L- and S-MAG upon cAMP stimulated differentiation in oligodendroglial cells. J Neurosci Res 2003; 71: 326–337.
Pellegrin S, Mellor H . Rho GTPase activation assays. Curr Protoc Cell Biol 2008 chapter 14 Unit 14, 18 doi:10.1002/0471143030.cb1408s38.
MacGillavry HD, Stam FJ, Sassen MM, Kegel L, Hendriks WT, Verhaagen J et al. NFIL3 and cAMP response element-binding protein form a transcriptional feedforward loop that controls neuronal regeneration-associated gene expression. J Neurosci 2009; 29: 15542–15550.
Acknowledgements
This work was supported by the Swiss National Science Foundation (SNF grant #31003A-149315-1 for MES), the European Research Council (ERC, ‘Nogorise’ #294115 for MES) and by the International Foundation for Research in Paraplegia (IRP, P115 for BT). We thank Professor Klaus-Armin Nave for kindly providing the Cnp-Cre mouse line, Professor Ronald van Kesteren for sending us the F11 cells and Professor Schaeren-Wiemers for sharing the L-MAG antiserum with us. We also thank A Guzik-Kornacka for help with tissue dissections and protein/mRNA extractions.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Edited by N Bazan
Supplementary Information accompanies this paper on Cell Death and Differentiation website
Supplementary information
Rights and permissions
About this article
Cite this article
Vajda, F., Jordi, N., Dalkara, D. et al. Cell type-specific Nogo-A gene ablation promotes axonal regeneration in the injured adult optic nerve. Cell Death Differ 22, 323–335 (2015). https://doi.org/10.1038/cdd.2014.147
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cdd.2014.147
This article is cited by
-
Synaptic or Non-synaptic? Different Intercellular Interactions with Retinal Ganglion Cells in Optic Nerve Regeneration
Molecular Neurobiology (2022)
-
Nogo-A-targeting antibody promotes visual recovery and inhibits neuroinflammation after retinal injury
Cell Death & Disease (2020)
-
Nerve Growth Factor Role on Retinal Ganglion Cell Survival and Axon Regrowth: Effects of Ocular Administration in Experimental Model of Optic Nerve Injury
Molecular Neurobiology (2019)
-
Nogo-A inactivation improves visual plasticity and recovery after retinal injury
Cell Death & Disease (2018)
-
Fasudil, a Rho-Associated Protein Kinase Inhibitor, Attenuates Traumatic Retinal Nerve Injury in Rabbits
Journal of Molecular Neuroscience (2016)
