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Urokinase-Type Plasminogen Activator Induces BV-2 Microglial Cell Migration Through Activation of Matrix Metalloproteinase-9

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Abstract

In response to brain injury, microglia migrate and accumulate in the affected sites, which is an important step in the regulation of inflammation and neuronal degeneration/regeneration. In this study, we investigated the effect of urokinase-type plasminogen activator (uPA) on the BV-2 microglial cell migration. At resting state, BV-2 microglial cells secreted uPA and the release of uPA was increased by ATP, a chemoattractant released from injured neuron. The migration of BV-2 cell was significantly induced by uPA and inhibited by uPA inhibitors. In this condition, uPA increased the activity of matrix metalloproteinase (MMP-9) and the inhibition of MMP activity with pharmacological inhibitors against either uPA (amiloride) or MMP (phenanthrolene and SB-3CT) effectively prevented BV2 cell migration. Interestingly, the level of MMP-9 protein and mRNA in the cell were not changed by uPA. These results suggest that the increase of MMP-9 activity by uPA is regulated at the post-translational level, possibly via increased activation of the enzyme. Unlike the uPA inhibitor, plasmin inhibitor PAI-1 only partially inhibited uPA-induced cell migration and MMP-9 activation. The incubation of recombinant MMP-9 with uPA resulted in the activation of MMP-9. These results suggest that uPA plays a critical role in BV-2 microglial cell migration by activating pro-MMP-9, in part by its direct action on MMP-9 and also in part by the activation of plasminogen/plasmin cascade.

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References

  1. Alfano D, Franco P, Vocca I, Gambi N, Pisa V, Mancini A, Caputi M, Carriero MV, Iaccarino I, Stoppelli MP (2005) The urokinase plasminogen activator and its receptor: role in cell growth and apoptosis. Thromb Haemost 93:205–211

    CAS  PubMed  Google Scholar 

  2. Baramova EN, Bajou K, Remacle A, L’Hoir C, Krell HW, Weidle UH, Noel A, Foidart JM (1997) Involvement of PA/plasmin system in the processing of pro-MMP-9 and in the second step of pro-MMP-2 activation. FEBS Lett 405:157–162

    Article  CAS  PubMed  Google Scholar 

  3. Blasi E, Barluzzi R, Bocchini V, Mazzolla R, Bistoni F (1990) Immortalization of murine microglial cells by a v-raf/v-myc carrying retrovirus. J Neuroimmunol 27:229–237

    Article  CAS  PubMed  Google Scholar 

  4. Blasi F (1993) Molecular mechanisms of protease-mediated tumor invasiveness. J Surg Oncol Suppl 3:21–23

    Article  CAS  PubMed  Google Scholar 

  5. Brockhaus J, Moller T, Kettenmann H (1996) Phagocytozing ameboid microglial cells studied in a mouse corpus callosum slice preparation. Glia 16:81–90

    Article  CAS  PubMed  Google Scholar 

  6. Carbonell WS, Murase S, Horwitz AF, Mandell JW (2005) Migration of perilesional microglia after focal brain injury and modulation by CC chemokine receptor 5: an in situ time-lapse confocal imaging study. J Neurosci 25:7040–7047

    Article  CAS  PubMed  Google Scholar 

  7. Castellino FJ, Ploplis VA (2005) Structure and function of the plasminogen/plasmin system. Thromb Haemost 93:647–654

    CAS  PubMed  Google Scholar 

  8. Clark AW, Krekoski CA, Bou SS, Chapman KR, Edwards DR (1997) Increased gelatinase A (MMP-2) and gelatinase B (MMP-9) activities in human brain after focal ischemia. Neurosci Lett 238:53–56

    Article  CAS  PubMed  Google Scholar 

  9. Dalmau I, Vela JM, Gonzalez B, Finsen B, Castellano B (2003) Dynamics of microglia in the developing rat brain. J Comp Neurol 458:144–157

    Article  PubMed  Google Scholar 

  10. DeClerck YA, Laug WE (1996) Cooperation between matrix metalloproteinases and the plasminogen activator-plasmin system in tumor progression. Enzyme Protein 49:72–84

    CAS  PubMed  Google Scholar 

  11. Elkabes S, DiCicco-Bloom EM, Black IB (1996) Brain microglia/macrophages express neurotrophins that selectively regulate microglial proliferation and function. J Neurosci 16:2508–2521

    CAS  PubMed  Google Scholar 

  12. Galko MJ, Tessier-Lavigne M (2000) Function of an axonal chemoattractant modulated by metalloprotease activity. Science 289:1365–1367

    Article  CAS  PubMed  Google Scholar 

  13. Gottschall PE, Yu X, Bing B (1995) Increased production of gelatinase B (matrix metalloproteinase-9) and interleukin-6 by activated rat microglia in culture. J Neurosci Res 42:335–342

    Article  CAS  PubMed  Google Scholar 

  14. Hahn-Dantona E, Ruiz JF, Bornstein P, Strickland DK (2001) The low density lipoprotein receptor-related protein modulates levels of matrix metalloproteinase 9 (MMP-9) by mediating its cellular catabolism. J Biol Chem 276:15498–15503

    Article  CAS  PubMed  Google Scholar 

  15. Honda S, Sasaki Y, Ohsawa K, Imai Y, Nakamura Y, Inoue K, Kohsaka S (2001) Extracellular ATP or ADP induce chemotaxis of cultured microglia through Gi/o-coupled P2Y receptors. J Neurosci 21:1975–1982

    CAS  PubMed  Google Scholar 

  16. Irigoyen JP, Munoz-Canoves P, Montero L, Koziczak M, Nagamine Y (1999) The plasminogen activator system: biology and regulation. Cell Mol Life Sci 56:104–132

    Article  CAS  PubMed  Google Scholar 

  17. Joo SH, Kwon KJ, Kim JW, Kim JW, Hasan MR, Lee HJ, Han SH, Shin CY (2010) Regulation of matrix metalloproteinase-9 and tissue plasminogen activator activity by alpha-synuclein in rat primary glial cells. Neurosci Lett 469:352–356

    Article  CAS  PubMed  Google Scholar 

  18. Kim J-A, Kang Y-R, Thapa D, Lee J-S, Park M-A, Lee K-H, Lyoo W-S, Lee Y-R (2009) Anti-invasive and anti-angiogenic effects of xanthohumol and its synthetic derivatives. Biomol Ther 17:422–429

    Article  CAS  Google Scholar 

  19. Kim J-S, Moon A-R (2009) Fibronectin induces pro-MMP-2 activation and enhances invasion in H-ras-transformed human breast epithelial cells. Biomol Ther 17:288–292

    CAS  Google Scholar 

  20. Kjoller L, Hall A (2001) Rac mediates cytoskeletal rearrangements and increased cell motility induced by urokinase-type plasminogen activator receptor binding to vitronectin. J Cell Biol 152:1145–1157

    Article  CAS  PubMed  Google Scholar 

  21. Kreutzberg GW (1996) Microglia: a sensor for pathological events in the CNS. Trends Neurosci 19:312–318

    Article  CAS  PubMed  Google Scholar 

  22. Kunigal S, Lakka SS, Gondi CS, Estes N, Rao JS (2007) RNAi-mediated downregulation of urokinase plasminogen activator receptor and matrix metalloprotease-9 in human breast cancer cells results in decreased tumor invasion, angiogenesis and growth. Int J Cancer 121:2307–2316

    Article  CAS  PubMed  Google Scholar 

  23. Kurpius D, Nolley EP, Dailey ME (2007) Purines induce directed migration and rapid homing of microglia to injured pyramidal neurons in developing hippocampus. Glia 55:873–884

    Article  PubMed  Google Scholar 

  24. Lee SY, Kim HJ, Lee WJ, Joo SH, Jeon SJ, Kim JW, Kim HS, Han SH, Lee J, Park SH, Cheong JH, Kim WK, Ko KH, Shin CY (2008) Differential regulation of matrix metalloproteinase-9 and tissue plasminogen activator activity by the cyclic-AMP system in lipopolysaccharide-stimulated rat primary astrocytes. Neurochem Res 33:2324–2334

    Article  CAS  PubMed  Google Scholar 

  25. Lee WJ, Shin CY, Yoo BK, Ryu JR, Choi EY, Cheong JH, Ryu JH, Ko KH (2003) Induction of matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide-stimulated primary astrocytes is mediated by extracellular signal-regulated protein kinase 1/2 (Erk1/2). Glia. 41:15–24

    Article  PubMed  Google Scholar 

  26. Liang X, Yang X, Tang Y, Zhou H, Liu X, Xiao L, Gao J, Mao Z (2008) RNAi-mediated downregulation of urokinase plasminogen activator receptor inhibits proliferation, adhesion, migration and invasion in oral cancer cells. Oral Oncol 44:1172–1180

    Article  CAS  PubMed  Google Scholar 

  27. Lorenzl S, Calingasan N, Yang L, Albers DS, Shugama S, Gregorio J, Krell HW, Chirichigno J, Joh T, Beal MF (2004) Matrix metalloproteinase-9 is elevated in 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine-induced parkinsonism in mice. Neuromolecular Med 5:119–132

    Article  CAS  PubMed  Google Scholar 

  28. Mazzieri R, Masiero L, Zanetta L, Monea S, Onisto M, Garbisa S, Mignatti P (1997) Control of type IV collagenase activity by components of the urokinase-plasmin system: a regulatory mechanism with cell-bound reactants. EMBO J 16:2319–2332

    Article  CAS  PubMed  Google Scholar 

  29. Nakajima K, Kohsaka S (2001) Microglia: activation and their significance in the central nervous system. J Biochem 130:169–175

    CAS  PubMed  Google Scholar 

  30. Nakajima K, Tohyama Y, Kurihara T, Kohsaka S (2005) Enhancement of urokinase-type plasminogen activator (uPA) secretion, but not that of substrate plasminogen (PGn), by rat microglia stimulated with neuronal conditioned medium. Neurosci Lett 378:13–17

    Article  CAS  PubMed  Google Scholar 

  31. Nakajima K, Tsuzaki N, Shimojo M, Hamanoue M, Kohsaka S (1992) Microglia isolated from rat brain secrete a urokinase-type plasminogen activator. Brain Res 577:285–292

    Article  CAS  PubMed  Google Scholar 

  32. Oh J, Takahashi R, Kondo S, Mizoguchi A, Adachi E, Sasahara RM, Nishimura S, Imamura Y, Kitayama H, Alexander DB, Ide C, Horan TP, Arakawa T, Yoshida H, Nishikawa S, Itoh Y, Seiki M, Itohara S, Takahashi C, Noda M (2001) The membrane-anchored MMP inhibitor RECK is a key regulator of extracellular matrix integrity and angiogenesis. Cell 107:789–800

    Article  CAS  PubMed  Google Scholar 

  33. Park GH, Jeon SJ, Ko HM, Ryu JR, Lee JM, Kim HY, Han SH, Kang YS, Park SH, Shin CY, Ko KH (2010) Activation of microglial cells via protease-activated receptor 2 mediates neuronal cell death in cultured rat primary neuron. Nitric Oxide 22:18–29

    Article  PubMed  CAS  Google Scholar 

  34. Ramos-Desimone N, Hahn-Dantona E, Sipley J, Nagase H, Debrah LF, James PQ (1999) Activation of Matrix Metalloproteinase-9(MMP-9) via a converging plasmin/stromelysin-1 cascade enhances tumor cell invasion. J Biol ChemV 274:13066–13076

    Article  CAS  Google Scholar 

  35. Rosenberg GA, Cunningham LA, Wallace J, Alexander S, Estrada EY, Grossetete M, Razhagi A, Miller K, Gearing A (2001) Immunohistochemistry of matrix metalloproteinases in reperfusion injury to rat brain: activation of MMP-9 linked to stromelysin-1 and microglia in cell cultures. Brain Res 893:104–112

    Article  CAS  PubMed  Google Scholar 

  36. Sidenius N, Blasi F (2003) The urokinase plasminogen activator system in cancer: recent advances and implication for prognosis and therapy. Cancer Metastasis Rev 22:205–222

    Article  CAS  PubMed  Google Scholar 

  37. Simard AR, Soulet D, Gowing G, Julien JP, Rivest S (2006) Bone marrow-derived microglia play a critical role in restricting senile plaque formation in Alzheimer’s disease. Neuron 49:489–502

    Article  CAS  PubMed  Google Scholar 

  38. Toth M, Sado Y, Ninomiya Y, Fridman R (1999) Biosynthesis of alpha2(IV) and alpha1(IV) chains of collagen IV and interactions with matrix metalloproteinase-9. J Cell Physiol 180:131–139

    Article  CAS  PubMed  Google Scholar 

  39. Vaillant C, Meissirel C, Mutin M, Belin MF, Lund LR, Thomasset N (2003) MMP-9 deficiency affects axonal outgrowth, migration, and apoptosis in the developing cerebellum. Mol Cell Neurosci 24:395–408

    Article  CAS  PubMed  Google Scholar 

  40. Walker DG, Lue LF, Beach TG (2002) Increased expression of the urokinase plasminogen-activator receptor in amyloid beta peptide-treated human brain microglia and in AD brains. Brain Res 926:69–79

    Article  CAS  PubMed  Google Scholar 

  41. Washington RA, Becher B, Balabanov R, Antel J, Dore-Duffy P (1996) Expression of the activation marker urokinase plasminogen-activator receptor in cultured human central nervous system microglia. J Neurosci Res 45:392–399

    Article  CAS  PubMed  Google Scholar 

  42. Yin KJ, Cirrito JR, Yan P, Hu X, Xiao Q, Pan X, Bateman R, Song H, Hsu FF, Turk J, Xu J, Hsu CY, Mills JC, Holtzman DM, Lee JM (2006) Matrix metalloproteinases expressed by astrocytes mediate extracellular amyloid-beta peptide catabolism. J Neurosci 26:10939–10948

    Article  CAS  PubMed  Google Scholar 

  43. Yong VW, Krekoski CA, Forsyth PA, Bell R, Edwards DR (1998) Matrix metalloproteinases and diseases of the CNS. Trends Neurosci 21:75–80

    Article  CAS  PubMed  Google Scholar 

  44. Zhao Y, Lyons CE Jr, Xiao A, Templeton DJ, Sang QA, Brew K, Hussaini IM (2008) Urokinase directly activates matrix metalloproteinases-9: a potential role in glioblastoma invasion. Biochem Biophys Res Commun 369:1215–1220

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This research was supported in part by a grant (M103KV010025-06K2201-02510) from Brain Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology, the Republic of Korea (K.H. Ko) and also in part by a grant (IBST-2008-01-02) from IBST, Konkuk University (C. Y. Shin).

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Authors and Affiliations

  1. Department of Pharmacology, College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, San 56-1, Shillim-Dong, Kwanak-Gu, Seoul, 151-742, Korea

    Sun Mi Shin, Kyu Suk Cho, Min Sik Choi & Kwang Ho Ko

  2. Institute for Biomedical Sciences and Technology, Konkuk University, Seoul, Korea

    Kyu Suk Cho, Sung Hoon Lee, Seol-Heui Han, Young-Sun Kang & Chan Young Shin

  3. Department of Pharmacology, School of Medicine, Konkuk University, Hwayang-Dong, Kwangjin-Gu, Seoul, 143-701, Korea

    Sung Hoon Lee & Chan Young Shin

  4. Department of Psychiatry, School of Medicine, Konkuk University, Seoul, Korea

    Seol-Heui Han

  5. Department of Biomedical Sciences and Technology, Konkuk University, Seoul, Korea

    Young-Sun Kang

  6. Department of Pharmacology, School of Pharmacy, Samyook University, Seoul, Korea

    Hee Jin Kim & Jae Hoon Cheong

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  1. Sun Mi Shin
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  9. Chan Young Shin
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Correspondence to Chan Young Shin or Kwang Ho Ko.

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11064_2010_141_MOESM1_ESM.pdf

Supplemental Figure 1. The effect of uPA on MMP-2 activity. The activity of MMP-2 released from BV-2 microglial cells into the culture supernatants was measured by gelatin zymography. Cultured BV-2 microglial cells were washed with serum-free DMEM medium and then treated with amiloride (100 μM) or tPA stop (2 μM) in the absence or presence of uPA (500 ng/ml) for 3 h at 37℃. Data are presented as the mean ± S.E.M. of four independent experiments. No siginificant difference was observed on MMP-2 activity. (PDF 25 kb)

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Shin, S.M., Cho, K.S., Choi, M.S. et al. Urokinase-Type Plasminogen Activator Induces BV-2 Microglial Cell Migration Through Activation of Matrix Metalloproteinase-9. Neurochem Res 35, 976–985 (2010). https://doi.org/10.1007/s11064-010-0141-3

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