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J Brachial Plex Peripher Nerve Inj 2008; 03(01): e58-e68
DOI: 10.1186/1749-7221-3-19
Research article
Fuentes et al; licensee BioMed Central Ltd.

Rho kinase inhibitors Y27632 and H1152 augment neurite extension in the presence of cultured Schwann cells[*]

Erick O Fuentes
1   Department of Plastic and Hand Surgery, University of Freiburg Medical Centre, Freiburg, Germany
,
Jost Leemhuis
2   Institute of Experimental and Clinical Pharmacology and Toxicology, Centre for Neuroscience, Freiburg, Germany
,
G Björn Stark
1   Department of Plastic and Hand Surgery, University of Freiburg Medical Centre, Freiburg, Germany
,
Eva M Lang
1   Department of Plastic and Hand Surgery, University of Freiburg Medical Centre, Freiburg, Germany
› Author Affiliations
Subject Editor:
Further Information

Publication History

08 April 2008

25 September 2008

Publication Date:
17 September 2014 (online)

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Abstract

Background RhoA and Rho kinase inhibitors overcome the inhibition of axonal regeneration posed by central nervous system (CNS) substrates.

Methods To investigate if inhibition of the Rho pathway augments the neurite extension that naturally occurs in the peripheral nervous system (PNS) following nerve damage, dorsal root ganglion neurons and Schwann cell co-cultures were incubated with culture medium, C3 fusion toxin, and the Rho kinase (ROCK) inhibitors Y27632 and H1152. The longest neurite per neuron were measured and compared. Incubation with Y27632 and H1152 resulted in significantly longer neurites than controls when the neurons were in contact with Schwann cells. When separated by a porous P.E.T. membrane, only the group incubated with H1152 developed significantly longer neurites. This work demonstrates that Rho kinase inhibition augments neurite elongation in the presence of contact with a PNS-like substrate.

*This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


 

  • References

  • 1 Giniger E. How do Rho family GTPases direct axon growth and guidance? A proposal relating signaling pathways to growth cone mechanics. Differentiation 2002; 70: 385-396 12366376
    CrossrefPubMedGoogle Scholar
  • 2 Amano M, Ito M, Kimura K, Fukata Y, Chihara K, Nakano T, Matsuura Y, Kaibuchi K. Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase). J Biol Chem 1996; 271: 20246-20249 8702756
    CrossrefPubMedGoogle Scholar
  • 3 Nobes CD, Hall A. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell 1995; 81: 53-62 7536630
    CrossrefPubMedGoogle Scholar
  • 4 Mackay DJ, Nobes CD, Hall A. The Rho’s progress: a potential role during neuritogenesis for the Rho family of GTPases. Trends Neurosci 1995; 18: 496-501 8592759
    CrossrefPubMedGoogle Scholar
  • 5 Sung JK, Miao L, Calvert JW, Huang L, Louis HH, Zhang JH. A possible role of RhoA/Rho-kinase in experimental spinal cord injury in rat. Brain Research 2003; 959: 29-38 12480155
    CrossrefPubMedGoogle Scholar
  • 6 Monnier PP, Sierra A, Schwab JM, Henke-Fahle S, Mueller BK. The Rho/ROCK pathway mediates neurite growth-inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar. Mol Cell Neurosci 2003; 22 (3) 319-330 12691734
    CrossrefPubMedGoogle Scholar
  • 7 Niederost B, Oertle T, Fritsche J, McKinney RA, Bandtlow CE. Nogo-A and myelin-associated glycoprotein mediate neurite growth inhibition by antagonistic regulation of RhoA and Rac1. J Neurosci 2002; 22: 10368-10376 12451136
    PubMedGoogle Scholar
  • 8 Bandtlow CE. Regeneration in the central nervous system. Experimental Gerontology 2003; 38: 79-86 12543264
    CrossrefPubMedGoogle Scholar
  • 9 Yamashita T, Tucker KL, Barde YA. Neurotrophin binding to the p75 receptor modulates Rho activity and axonal outgrowth. Neuron 1999; 24: 585-593 10595511
    CrossrefPubMedGoogle Scholar
  • 10 Yamashita T, Tohyama M. The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI. Nat Neurosci 2003; 6: 461-467 12692556
    PubMedGoogle Scholar
  • 11 Gehler S, Gallo G, Veien E, Letourneau PC. p75 neurotrophin receptor signaling regulates growth cone filopodial dynamics through modulating RhoA activity. J Neurosci 2004; 24: 4363-4372 15128850
    CrossrefPubMedGoogle Scholar
  • 12 Mi S, Lee X, Shao Z, Thill G, Ji B, Relton J, Levesque M, Allaire N, Perrin S, Sands B, Crowell T, Cate RL, McCoy JM, Pepinsky RB. LINGO-1 is a component of the Nogo-66 receptor/p75 signaling complex. Nat Neurosci 2004; 7: 221-228 14966521
    CrossrefPubMedGoogle Scholar
  • 13 Terashima T, Yasuda H, Terada M, Kogawa S, Maeda K, Haneda M, Kashiwagi A, Kikkawa R. Expression of Rho-family GTPases (Rac, cdc42, RhoA) and their association with p-21 activated kinase in adult rat peripheral nerve. J Neurochem 2001; 77: 986-993 11359863
    CrossrefPubMedGoogle Scholar
  • 14 Hiraga A, Kuwabara S, Doya H, Kanai K, Fujitani M, Taniguchi J, Arai K, Mori M, Hattori T, Yamashita T. Rho-kinase inhibition enhances axonal regeneration after peripheral nerve injury. J Peripher Nerv Syst 2006; 11: 217-224 16930283
    CrossrefPubMedGoogle Scholar
  • 15 Melendez-Vasquez CV, Einheber S, Salzer JL. Rho kinase regulates schwann cell myelination and formation of associated axonal domains. J Neurosci 2004; 24: 3953-3963 15102911
    CrossrefPubMedGoogle Scholar
  • 16 Fernandez-Valle C, Wood PM, Bunge MB. Localization of focal adhesion kinase in differentiating Schwann cell/neuron cultures. Microsc Res Tech 1998; 41 (5) 416-430 9672424
    CrossrefPubMedGoogle Scholar
  • 17 Yamauchi J, Chan JR, Shooter EM. Neurotrophins regulate Schwann cell migration by activating divergent signaling pathways dependent on Rho GTPases. Proc Natl Acad Sci USA 2004; 101: 8774-8779 423271 15161978
    CrossrefPubMedGoogle Scholar
  • 18 Barth H, Blocker D, Aktories K. The uptake machinery of clostridial actin ADP-ribosylating toxins – a cell delivery system for fusion proteins and polypeptide drugs. Naunyn Schmiedebergs Arch Pharmacol 2002; 366: 501-512 12444490
    CrossrefPubMedGoogle Scholar
  • 19 Ishizaki T, Uehata M, Tamechika I, Keel J, Nonomura K, Maekawa M, Narumiya S. Pharmacological properties of Y-2 a specific inhibitor of rho-associated kinases. Mol Pharmacol 7632, 57: 976-983
    PubMedGoogle Scholar
  • 20 Uehata M, Ishizaki T, Satoh H, Ono T, Kawahara T, Morishita T, Tamakawa H, Yamagami K, Inui J, Maekawa M, Narumiya S. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 1997; 389: 990-994 9353125
    CrossrefPubMedGoogle Scholar
  • 21 Ikenoya M, Hidaka H, Hosoya T, Suzuki M, Yamamoto N, Sasaki Y. Inhibition of rho-kinase-induced myristoylated alanine-rich C kinase substrate (MARCKS) phosphorylation in human neuronal cells by H- a novel and specific Rho-kinase inhibitor. J Neurochem 1152; 81: 9-16
    PubMedGoogle Scholar
  • 22 Sasaki Y, Suzuki M, Hidaka H. The novel and specific Rho-kinase inhibitor (S)-(+)-2-methyl-1-[(4-methyl-5-isoquinoline)sulfonyl]-homopiperazine as a probing molecule for Rho-kinase-involved pathway. Pharmacol Ther 2002; 93: 225-232 12191614
    CrossrefPubMedGoogle Scholar
  • 23 Breitenlechner C, Gassel M, Hidaka H, Kinzel V, Huber R, Engh RA, Bossemeyer D. Protein kinase A in complex with Rho-kinase inhibitors Y-27632 Fasudil, and H-1152P: structural basis of selectivity. Structure 2003; 11 (12) 1595-1607 14656443
    CrossrefPubMedGoogle Scholar
  • 24 Seilheimer B, Persohn E, Schachner M. Neural cell adhesion molecule expression is regulated by Schwann cell-neuron interactions in culture. Journal of Cell Biology 1989; 108: 1909-1915 2115570 2715182
    CrossrefPubMedGoogle Scholar
  • 25 Henle F, Leemhuis J, Fischer C, Bock HH, Lindemeyer K, Feuerstein TJ, Meyer DK. Gabapentin-lactam induces dendritic filopodia and motility in cultured hippocampal neurons. J Pharmacol Exp Ther 2006; 319: 181-191 16844845
    CrossrefPubMedGoogle Scholar
  • 26 Henle F, Fischer C, Meyer DK, Leemhuis J. Vasoactive intestinal peptide and PACAP38 control N-methyl-D-aspartic acid-induced dendrite motility by modifying the activities of Rho GTPases and phosphatidylinositol 3-kinases. J Biol Chem 2006; 281: 24955-24969 16803895
    CrossrefPubMedGoogle Scholar
  • 27 Lehmann M, Fournier A, Selles-Navarro I, Dergham P, Sebok A, Leclerc N, Tigyi G, McKerracher L. Inactivation of Rho signaling pathway promotes CNS axon regeneration. J Neurosci 1999; 19: 7537-7547 10460260
    CrossrefPubMedGoogle Scholar
  • 28 Thies E, Davenport RW. Independent roles of Rho-GTPases in growth cone and axonal behavior. Journal of Neurobiology 2003; 54: 358-369 12500311
    CrossrefPubMedGoogle Scholar
  • 29 Ellezam B, Dubreuil C, Winton M, Loy L, Dergham P. Inactivation of intracellular Rho to stimulate axon growth and regeneration. Progress in Brain Research 2002; 137: 371-380 12440379
    PubMedGoogle Scholar
  • 30 Borisoff JF, Chan CC, Hiebert GW, Oschipok L, Robertson GS. Suppression of Rho-kinase activity promotes axonal growth on inhibitory CNS substrates. Mol Cell Neurosci 2003; 22 (3) 405-416 12691741
    CrossrefPubMedGoogle Scholar
  • 31 Goldberg JL, Barres BA. Nogo in nerve regeneration. Nature 2000; 403: 369-370 10667770
    CrossrefPubMedGoogle Scholar
  • 32 Sandvig A, Berry M, Barrett LB, Butt A, Logan A. Myelin-, reactive glia-, and scar-derived CNS axon growth inhibitors: expression, receptor signaling, and correlation with axon regeneration. Glia 2004; 46: 225-251 15048847
    CrossrefPubMedGoogle Scholar
  • 33 McGee AW, Strittmatter SM. The Nogo-66 receptor: focusing myelin inhibition of axon regeneration. Trends in Neurosciences 2003; 26: 193-198 12689770
    CrossrefPubMedGoogle Scholar
  • 34 Schwab ME. Nogo and axon regeneration. Current Opinion in Neurobiology 2004; 14: 118-124 15018947
    CrossrefPubMedGoogle Scholar
  • 35 Fu SY, Gordon T. The cellular and molecular basis of peripheral nerve regeneration. Mol Neurobiol 1997; 14: 67-116 9170101
    CrossrefPubMedGoogle Scholar
  • 36 Bunge MB, Bunge RP, Kleitman N, Dean AC. Role of peripheral nerve extracellular matrix in Schwann cell function and in neurite regeneration. Dev Neurosci 1989; 11: 348-360 2676458
    CrossrefPubMedGoogle Scholar
  • 37 Bunge RP. The role of the Schwann cell in trophic support and regeneration. J Neurol 1994; 242: S19-S21 7699403
    CrossrefPubMedGoogle Scholar
  • 38 Meyer M, Matsuoka I, Wetmore C, Olson L, Thoenen H. Enhanced synthesis of brain-derived neurotrophic factor in the lesioned peripheral nerve: different mechanisms are responsible for the regulation of BDNF and NGF mRNA. J Cell Biol 1992; 119: 45-54 2289627 1527172
    CrossrefPubMedGoogle Scholar
  • 39 Hempstead BL. The many faces of p75NTR. Curr Opin Neurobiol 2002; 12: 260-267 12049931
    CrossrefPubMedGoogle Scholar
  • 40 Chao MV. Neurotrophins and their receptors: a convergence point for many signalling pathways. Nat Rev Neurosci 2003; 4: 299-309 12671646
    CrossrefPubMedGoogle Scholar
  • 41 Kobayashi K, Takahashi M, Matsushita N, Miyazaki J, Koike M. Survival of developing motor neurons mediated by Rho GTPase signaling pathway through Rho-kinase. J Neurosci 2004; 24: 3480-3488 15071095
    CrossrefPubMedGoogle Scholar
  • 42 Seilheimer B, Schachner M. Regulation of neural cell adhesion molecule expression on cultured mouse Schwann cells by nerve growth factor. EMBO J 1987; 6: 1611-1616 553532 3608988
    PubMedGoogle Scholar
  • 43 Seilheimer B, Schachner M. Studies of adhesion molecules mediating interactions between cells of peripheral nervous system indicate a major role for L1 in mediating sensory neuron growth on Schwann cells in culture. Journal of Cell Biology 1988; 107: 341-351 2115159 3292543
    CrossrefPubMedGoogle Scholar
  • 44 Haney CA, Sahenk Z, Li C, Lemmon VP, Roder J, Trapp BD. Heterophilic binding of L1 on unmyelinated sensory axons mediates Schwann cell adhesion and is required for axonal survival. Journal of Cell Biology 1999; 146: 1173-1184 2169489 10477768
    CrossrefPubMedGoogle Scholar
  • 45 David S, Braun PE, Jackson DL, Kottis V, McKerracher L, David S, Braun PE, Jackson DL, Kottis V, McKerracher L. Laminin overrides the inhibitory effects of peripheral nervous system and central nervous system myelin-derived inhibitors of neurite growth. Journal of Neuroscience Research 1995; 42: 594-602 8568945
    CrossrefPubMedGoogle Scholar
  • 46 Bruck W, Bruck Y, Maruschak B, Friede RL. Mechanisms of macrophage recruitment in Wallerian degeneration. Acta Neuropathol 1995; 89: 363-367 7610768
    CrossrefPubMedGoogle Scholar
  • 47 Stoll G, Griffin JW, Li CY, Trapp BD. Wallerian degeneration in the peripheral nervous system: participation of both Schwann cells and macrophages in myelin degradation. J Neurocytol 1989; 18: 671-683 2614485
    CrossrefPubMedGoogle Scholar
  • 48 Cai D, Qiu J, Cao Z, McAtee M, Bregman BS, Filbin MT. Neuronal cyclic AMP controls the developmental loss in ability of axons to regenerate. J Neurosci 2001; 21: 4731-4739 11425900
    PubMedGoogle Scholar
  • 49 Fukata M, Nakagawa M, Kuroda S, Kaibuchi K. Cell adhesion and Rho small GTPases. J Cell Sci 1999; 112 (Pt 24) 4491-4500 10574699
    PubMedGoogle Scholar