Spinal cord injury induction of lesional expression of profibrotic and angiogenic connective tissue growth factor confined to reactive astrocytes, invading fibroblasts and endothelial cells

Sabine Conrad T.A.1, Hermann J. Schluesener M.D., Ph.D.1, Mehdi Adibzahdeh Ph.D.1, and Jan M. Schwab M.D., Ph.D.1
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  • 1 Institute of Brain Research, University of Tübingen, Medical School, Tübingen, Germany; and Equipe Développement Neuronal, Université Pierre et Marie Curie, Paris, France
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Object. The glial scar composed of astrogliosis and extracellular matrix deposition represents a major impediment to axonal regeneration. The authors investigated the role of a novel profibrotic and angiogenic peptide connective tissue growth factor (CTGF [Hcs24/IGFBP-r2P]) in glial scar formation following spinal cord injury (SCI) in rats.

Methods. The effects of SCI on CTGF expression during glial scar maturation 1 day to 1 month post-SCI were investigated using fluorescein-activated cell sorter (FACS) immunohistochemical analysis; these findings were compared with those obtained in sham-operated (control) spinal cords.

The CTGF-positive cells accumulated at the spinal cord lesion site (p < 0.0001) corresponding to areas of glial scar formation. In the perilesional rim, CTGF expression was confined to invading vimentin-positive, glial fibrillary acidic protein (GFAP)—negative fibroblastoid cells, endothelial and smooth-muscle cells of laminin-positive vessels, and GFAP-positive reactive astrocytes. The CTGF-positive astrocytes coexpressed the activation-associated intermediate filaments nestin, vimentin (> 80%), and mesenchymal scar component fibronectin (50%).

Conclusions. The restricted accumulation of CTGF-reactive astrocytes and CTGF-positive fibroblastoid cells lining the laminin-positive basal neolamina suggests participation of these cells in scar formation. In addition, perilesional upregulation of endothelial and smooth-muscle CTGF expression points to a role in blood—brain barrier function modulating edema-induced secondary damage.

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  • 1.

    Babic AM, , Chen CC, & Lau LF: Fisp12/mouse connective tissue growth factor mediates endothelial cell adhesion and migration through integrin alphavbeta3, promotes endothelial cells survival, and induces angiogenesis in vivo. Mol Cell Biol 19:29582966, 1999 Babic AM, Chen CC, Lau LF: Fisp12/mouse connective tissue growth factor mediates endothelial cell adhesion and migration through integrin alphavbeta3, promotes endothelial cells survival, and induces angiogenesis in vivo. Mol Cell Biol 19:2958–2966, 1999

    • Search Google Scholar
    • Export Citation
  • 2.

    Bignami A: Glial types, gliogenesis, and extracellular matrix in the mammalian CNS, in Vernadakis A, & Roots B (eds): Neuron-Glia Interrelations During Phylogeny, ed 1. Totowa: Humana Press, 1998, pp 339 Bignami A: Glial types, gliogenesis, and extracellular matrix in the mammalian CNS, in Vernadakis A, Roots B (eds): Neuron-Glia Interrelations During Phylogeny, ed 1. Totowa: Humana Press, 1998, pp 3–39

    • Search Google Scholar
    • Export Citation
  • 3.

    Cancilla PA, , Bready J, & Berliner J: Astrocyte-endothelial cell interactions, in S Murphy (ed): Astrocytes. Pharmacology and Function. San Diego: Academic Press, 1993, pp 387397 Cancilla PA, Bready J, Berliner J: Astrocyte-endothelial cell interactions, in S Murphy (ed): Astrocytes. Pharmacology and Function. San Diego: Academic Press, 1993, pp 387–397

    • Search Google Scholar
    • Export Citation
  • 4.

    Dammeier J, , Brauchle M, , Falk W, , Grotendorst GR, & Werner S: Connective tissue growth factor: a novel regulator of mucosal repair and fibrosis in inflammatory bowel disease? Int J Biochem Cell Biol 30:909922, 1998 Dammeier J, Brauchle M, Falk W, Grotendorst GR, Werner S: Connective tissue growth factor: a novel regulator of mucosal repair and fibrosis in inflammatory bowel disease? Int J Biochem Cell Biol 30:909–922, 1998

    • Search Google Scholar
    • Export Citation
  • 5.

    Daniels JT, , Schultz GS, , Blalock TD, , Garrett Q, , Grotendorst GR, & Dean NM, et al: Mediation of transforming growth factorbeta(1)-stimulated matrix contraction by fibroblasts: a role for connective tissue growth factor in contractile scarring. Am J Pathol 163:20432052, 2003 Daniels JT, Schultz GS, Blalock TD, Garrett Q, Grotendorst GR, Dean NM, et al: Mediation of transforming growth factorbeta(1)-stimulated matrix contraction by fibroblasts: a role for connective tissue growth factor in contractile scarring. Am J Pathol 163:2043–2052, 2003

    • Search Google Scholar
    • Export Citation
  • 6.

    Duncan MR, , Frazier KS, , Abramson S, , Williams S, , Klapper H, & Huang X, et al: Connective tissue growth factor mediates transforming growth factor b-induced collagen synthesis: down-regulation by cAMP. FASEB J 13:17741786, 1999 Duncan MR, Frazier KS, Abramson S, Williams S, Klapper H, Huang X, et al: Connective tissue growth factor mediates transforming growth factor b-induced collagen synthesis: down-regulation by cAMP. FASEB J 13:1774–1786, 1999

    • Search Google Scholar
    • Export Citation
  • 7.

    Faden IF: Pharmacological treatment approaches for brain and spinal cord trauma, in Narayan RK, , Wilberger JE, & Povlishock JT (eds): Neurotrauma, ed 1. New York: McGraw-Hill, 1996, pp 14791490 Faden IF: Pharmacological treatment approaches for brain and spinal cord trauma, in Narayan RK, Wilberger JE, Povlishock JT (eds): Neurotrauma, ed 1. New York: McGraw-Hill, 1996, pp 1479–1490

    • Search Google Scholar
    • Export Citation
  • 8.

    Fawcett JW, & Asher RA: The glial scar and central nervous system repair. Brain Res Bull 49:377391, 1999 Fawcett JW, Asher RA: The glial scar and central nervous system repair. Brain Res Bull 49:377–391, 1999

    • Search Google Scholar
    • Export Citation
  • 9.

    Grotendorst GR: Connective tissue growth factor: a mediator of TGF-β action on fibroblasts. Cytokine Growth Factor Rev 8:171179, 1997 Grotendorst GR: Connective tissue growth factor: a mediator of TGF-β action on fibroblasts. Cytokine Growth Factor Rev 8:171–179, 1997

    • Search Google Scholar
    • Export Citation
  • 10.

    Hamada Y, , Ikata T, , Katoh S, , Katoh K, , Niwa K, & Tsutsumishita Y, et al: Effects of exogenous transforming growth factor-beta 1 on spinal cord injury in rats. Neurosci Lett 203:97100, 1996 Hamada Y, Ikata T, Katoh S, Katoh K, Niwa K, Tsutsumishita Y, et al: Effects of exogenous transforming growth factor-beta 1 on spinal cord injury in rats. Neurosci Lett 203:97–100, 1996

    • Search Google Scholar
    • Export Citation
  • 11.

    Hashimoto Y, , Shindo-Okada N, , Tani M, , Nagamachi Y, , Takeuchi Y, & Shiroishi T, et al: Expression of the Elm1 gene, a novel gene of the CCN (connective tissue growth factor, Cyr61/Cef10, and neuroblastoma overexpressed gene) family, suppresses in vivo tumor growth and metastasis of K-1735 murine melanoma cells. J Exp Med 187:289296, 1998 Hashimoto Y, Shindo-Okada N, Tani M, Nagamachi Y, Takeuchi Y, Shiroishi T, et al: Expression of the Elm1 gene, a novel gene of the CCN (connective tissue growth factor, Cyr61/Cef10, and neuroblastoma overexpressed gene) family, suppresses in vivo tumor growth and metastasis of K-1735 murine melanoma cells. J Exp Med 187:289–296, 1998

    • Search Google Scholar
    • Export Citation
  • 12.

    Henrich-Noack P, , Prehn JH, & Krieglstein J: TGF-β 1 protects hippocampal neurons against degeneration caused by transient global ischemia. Dose-response relationship and potential neuroprotective mechanisms. Stroke 27:16091615, 1996 Henrich-Noack P, Prehn JH, Krieglstein J: TGF-β 1 protects hippocampal neurons against degeneration caused by transient global ischemia. Dose-response relationship and potential neuroprotective mechanisms. Stroke 27:1609–1615, 1996

    • Search Google Scholar
    • Export Citation
  • 13.

    Heuer H, , Christ S, , Friedrichsen S, , Brauer D, , Winckler M, & Bauer K, et al: Connective tissue growth factor: a novel marker of layer VII neurons in the rat cerebral cortex. Neuroscience 119:4352, 2003 Heuer H, Christ S, Friedrichsen S, Brauer D, Winckler M, Bauer K, et al: Connective tissue growth factor: a novel marker of layer VII neurons in the rat cerebral cortex. Neuroscience 119:43–52, 2003

    • Search Google Scholar
    • Export Citation
  • 14.

    Hertel M, , Tretter Y, , Alzheimer C, & Werner S: Connective tissue growth factor: a novel player in tissue reorganization after brain injury? Eur J Neurosci 12:376380, 2000 Hertel M, Tretter Y, Alzheimer C, Werner S: Connective tissue growth factor: a novel player in tissue reorganization after brain injury? Eur J Neurosci 12:376–380, 2000

    • Search Google Scholar
    • Export Citation
  • 15.

    Imperato-Kalmar EL, , McKinney RA, , Schnell L, , Rubin BP, & Schwab ME: Local changes in vascular architecture following partial spinal cord lesion in the rat. Exp Neurol 145:322328, 1997 Imperato-Kalmar EL, McKinney RA, Schnell L, Rubin BP, Schwab ME: Local changes in vascular architecture following partial spinal cord lesion in the rat. Exp Neurol 145:322–328, 1997

    • Search Google Scholar
    • Export Citation
  • 16.

    Kim HS, , Nagalla SR, , Oh Y, , Wilson E, , Roberts CT Jr, & Rosenfeld RG: Identification of a family of low-affinity insulin-like growth factor binding proteins (IGFBPs): characterization of connective tissue growth factor as a member of the IGFBP superfamily. Proc Natl Acad Sci USA 94:1298112986, 1997 Kim HS, Nagalla SR, Oh Y, Wilson E, Roberts CT Jr, Rosenfeld RG: Identification of a family of low-affinity insulin-like growth factor binding proteins (IGFBPs): characterization of connective tissue growth factor as a member of the IGFBP superfamily. Proc Natl Acad Sci USA 94:12981–12986, 1997

    • Search Google Scholar
    • Export Citation
  • 17.

    Kondo Y, , Nakanishi T, , Takigawa M, & Ogawa N: Immunohistochemical localization of connective tissue growth factor in the rat central nervous system. Brain Res 834:146151, 1999 Kondo Y, Nakanishi T, Takigawa M, Ogawa N: Immunohistochemical localization of connective tissue growth factor in the rat central nervous system. Brain Res 834:146–151, 1999

    • Search Google Scholar
    • Export Citation
  • 18.

    Kubo M, , Kikuchi K, , Nashiro K, , Kakinuma T, , Hayashi N, & Nanko H, et al: Expression of fibrogenic cytokines in desmoplastic malignant melanoma. Br J Dermatol 139:192197, 1998 Kubo M, Kikuchi K, Nashiro K, Kakinuma T, Hayashi N, Nanko H, et al: Expression of fibrogenic cytokines in desmoplastic malignant melanoma. Br J Dermatol 139:192–197, 1998

    • Search Google Scholar
    • Export Citation
  • 19.

    Lasky JA, , Ortiz LA, , Tonthat B, , Hoyle GW, , Corti M, & Athas G, et al: Connective tissue growth factor mRNA expression is upregulated in bleomycin-induced lung fibrosis. Am J Physiol 275:L365L371, 1998 Lasky JA, Ortiz LA, Tonthat B, Hoyle GW, Corti M, Athas G, et al: Connective tissue growth factor mRNA expression is upregulated in bleomycin-induced lung fibrosis. Am J Physiol 275:L365–L371, 1998

    • Search Google Scholar
    • Export Citation
  • 20.

    Lawrence DA: Transforming growth factor-β: a general review. Eur Cytokine Netw 7:363374, 1996 Lawrence DA: Transforming growth factor-β: a general review. Eur Cytokine Netw 7:363–374, 1996

    • Search Google Scholar
    • Export Citation
  • 21.

    Logan A, & Berry M: Transforming growth factor-β1 and basic fibroblast growth factor in the injured CNS. Trends Pharmacol Sci 14:337342, 1993 Logan A, Berry M: Transforming growth factor-β1 and basic fibroblast growth factor in the injured CNS. Trends Pharmacol Sci 14:337–342, 1993

    • Search Google Scholar
    • Export Citation
  • 22.

    Logan A, , Berry M, , Gonzalez AM, , Frautschy SA, , Sporn MB, & Baird A: Effects of transforming growth factor β1 on scar production in the injured central nervous system of the rat. Eur J Neurosci 6:355363, 1994 Logan A, Berry M, Gonzalez AM, Frautschy SA, Sporn MB, Baird A: Effects of transforming growth factor β1 on scar production in the injured central nervous system of the rat. Eur J Neurosci 6:355–363, 1994

    • Search Google Scholar
    • Export Citation
  • 23.

    Martinerie C, , Viegas-Pequignot E, , Guenard I, , Dutrillaux B, , Nguyen VC, & Bernheim A, et al: Physical mapping of human loci homologous to the chicken nov proto-oncogene. Oncogene 7:25292534, 1992 Martinerie C, Viegas-Pequignot E, Guenard I, Dutrillaux B, Nguyen VC, Bernheim A, et al: Physical mapping of human loci homologous to the chicken nov proto-oncogene. Oncogene 7:2529–2534, 1992

    • Search Google Scholar
    • Export Citation
  • 24.

    McKeon RJ, , Schreiber RC, , Rudge JS, & Silver J: Reduction of neurite outgrowth in a model of glial scarring following CNS injury is correlated with the expression of inhibitory molecules on reactive astroytes. J Neurosci 11:33983411, 1991 McKeon RJ, Schreiber RC, Rudge JS, Silver J: Reduction of neurite outgrowth in a model of glial scarring following CNS injury is correlated with the expression of inhibitory molecules on reactive astroytes. J Neurosci 11:3398–3411, 1991

    • Search Google Scholar
    • Export Citation
  • 25.

    Merkler D, , Metz GA, , Raineteau O, , Dietz V, , Schwab ME, & Fouad K: Locomotor recovery in spinal cord-injured rats treated with an antibody neutralizing the myelin-associated neurite growth inhibitor Nogo-A. J Neurosci 21:36653673, 2001 Merkler D, Metz GA, Raineteau O, Dietz V, Schwab ME, Fouad K: Locomotor recovery in spinal cord-injured rats treated with an antibody neutralizing the myelin-associated neurite growth inhibitor Nogo-A. J Neurosci 21:3665–3673, 2001

    • Search Google Scholar
    • Export Citation
  • 26.

    Miller JD, & Ironside JW: Raised intracranial pressure, oedema and hydrocephalus, in Graham DI, & Lantos PL (eds): Greenfield's Neuropathology, ed 6. London: Arnold, 1996, pp157187 Miller JD, Ironside JW: Raised intracranial pressure, oedema and hydrocephalus, in Graham DI, Lantos PL (eds): Greenfield's Neuropathology, ed 6. London: Arnold, 1996, pp157–187

    • Search Google Scholar
    • Export Citation
  • 27.

    Nakanishi T, , Nishida T, , Shimo T, , Kobayashi K, , Kubo T, & Tamatani T, et al: Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture. Endocrinology 141:264273, 2000 Nakanishi T, Nishida T, Shimo T, Kobayashi K, Kubo T, Tamatani T, et al: Effects of CTGF/Hcs24, a product of a hypertrophic chondrocyte-specific gene, on the proliferation and differentiation of chondrocytes in culture. Endocrinology 141:264–273, 2000

    • Search Google Scholar
    • Export Citation
  • 28.

    Norenberg MD: Astrocyte responses to CNS injury. J Neuropathol Exp Neurol 53:213220, 1994 Norenberg MD: Astrocyte responses to CNS injury. J Neuropathol Exp Neurol 53:213–220, 1994

    • Search Google Scholar
    • Export Citation
  • 29.

    Oemar BS, , Werner A, , Garnier JM, , Do DD, , Godoy N, & Nauck M, et al: Human connective tissue growth factor is expressed in advanced atherosclerotic lesions. Circulation 95:831839, 1997 Oemar BS, Werner A, Garnier JM, Do DD, Godoy N, Nauck M, et al: Human connective tissue growth factor is expressed in advanced atherosclerotic lesions. Circulation 95:831–839, 1997

    • Search Google Scholar
    • Export Citation
  • 30.

    Schluesener HJ, , Seid K, & Meyermann R: Effects of autoantigen and dexamethasone treatment on expression of endothelial-monocyte activating polypeptide II and allograft-inflammatory factor-1 by activated macrophages and microglial cells in lesions of experimental autoimmune encephalomyelitits, neuritis and uveitis. Acta Neuropathol 97:119126, 1999 Schluesener HJ, Seid K, Meyermann R: Effects of autoantigen and dexamethasone treatment on expression of endothelial-monocyte activating polypeptide II and allograft-inflammatory factor-1 by activated macrophages and microglial cells in lesions of experimental autoimmune encephalomyelitits, neuritis and uveitis. Acta Neuropathol 97:119–126, 1999

    • Search Google Scholar
    • Export Citation
  • 31.

    Schnell L, , Fearn S, , Klassen H, , Schwab ME, & Perry VH: Acute inflammatory responses to mechanical lesions in the CNS: differences between brain and spinal cord. Eur J Neurosci 11:36483658, 1999 Schnell L, Fearn S, Klassen H, Schwab ME, Perry VH: Acute inflammatory responses to mechanical lesions in the CNS: differences between brain and spinal cord. Eur J Neurosci 11:3648–3658, 1999

    • Search Google Scholar
    • Export Citation
  • 32.

    Schwab ME, & Bartholdi D: Degeneration and regeneration of axons in the lesioned spinal cord. Physiol Rev 76:319370, 1996 Schwab ME, Bartholdi D: Degeneration and regeneration of axons in the lesioned spinal cord. Physiol Rev 76:319–370, 1996

    • Search Google Scholar
    • Export Citation
  • 33.

    Schwab JM, , Beschorner R, , Nguyen TD, , Meyermann R, & Schluesener HJ: Differential cellular accumulation of connective tissue growth factor defines a subset of reactive astrocytes, invading fibroblasts, and endothelial cells following central nervous system injury in rats and humans. J Neurotrauma 18:377388, 2001 Schwab JM, Beschorner R, Nguyen TD, Meyermann R, Schluesener HJ: Differential cellular accumulation of connective tissue growth factor defines a subset of reactive astrocytes, invading fibroblasts, and endothelial cells following central nervous system injury in rats and humans. J Neurotrauma 18:377–388, 2001

    • Search Google Scholar
    • Export Citation
  • 34.

    Schwab JM, , Postler E, , Nguyen TD, , Mittelbronn M, , Meyermann R, & Schluesener HJ: Connective tissue growth factor is expressed by a subset of reactive astrocytes in human cerebral infarction. Neuropathol Appl Neurobiol 26:434440, 2000 Schwab JM, Postler E, Nguyen TD, Mittelbronn M, Meyermann R, Schluesener HJ: Connective tissue growth factor is expressed by a subset of reactive astrocytes in human cerebral infarction. Neuropathol Appl Neurobiol 26:434–440, 2000

    • Search Google Scholar
    • Export Citation
  • 35.

    Silver J, & Miller JH: Regeneration beyond the glial scar. Nat Rev Neurosci 5:14656, 2004 Silver J, Miller JH: Regeneration beyond the glial scar. Nat Rev Neurosci 5:146–56, 2004

    • Search Google Scholar
    • Export Citation
  • 36.

    Steffen CL, , Ball-Mirth DK, , Harding PA, , Bhattacharyya N, , Pillai S, & Brigstock DR: Characterization of cell-associated and soluble forms of connective tissue growth factor (CTGF) produced by fibroblast cells in vitro. Growth Factors 15:199213, 1998 Steffen CL, Ball-Mirth DK, Harding PA, Bhattacharyya N, Pillai S, Brigstock DR: Characterization of cell-associated and soluble forms of connective tissue growth factor (CTGF) produced by fibroblast cells in vitro. Growth Factors 15:199–213, 1998

    • Search Google Scholar
    • Export Citation
  • 37.

    Stichel CC, & Muller HW: Experimental strategies to promote axonal regeneration after traumatic central nervous system injury. Prog Neurobiol 56:119148, 1998 Stichel CC, Muller HW: Experimental strategies to promote axonal regeneration after traumatic central nervous system injury. Prog Neurobiol 56:119–148, 1998

    • Search Google Scholar
    • Export Citation
  • 38.

    Tator CH: Update on the pathophysiology and pathology of acute spinal cord injury. Brain Pathol 5:407413, 1995 Tator CH: Update on the pathophysiology and pathology of acute spinal cord injury. Brain Pathol 5:407–413, 1995

    • Search Google Scholar
    • Export Citation
  • 39.

    Toru-Delbauffe D, , Baghdassarian D, , Both D, , Bernard R, , Rouget P, & Pierre M: Effects of TGF-β1 on the proliferation and differentiation of an immortalized astrocyte cell line: relationship with extracellular matrix. Exp Cell Res 202:316325, 1992 Toru-Delbauffe D, Baghdassarian D, Both D, Bernard R, Rouget P, Pierre M: Effects of TGF-β1 on the proliferation and differentiation of an immortalized astrocyte cell line: relationship with extracellular matrix. Exp Cell Res 202:316–325, 1992

    • Search Google Scholar
    • Export Citation
  • 40.

    Wenger C, , Ellenrieder V, , Alber B, , Lacher U, , Menke A, & Hameister H, et al: Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells. Oncogene 18:10731080, 1999 Wenger C, Ellenrieder V, Alber B, Lacher U, Menke A, Hameister H, et al: Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells. Oncogene 18:1073–1080, 1999

    • Search Google Scholar
    • Export Citation

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