Toll‐like receptor 4 is required for α‐synuclein dependent activation of microglia and astroglia

Abstract

Alpha‐synucleinopathies (ASP) are neurodegenerative disorders, characterized by accumulation of misfolded α‐synuclein, selective neuronal loss, and extensive gliosis. It is accepted that microgliosis and astrogliosis contribute to the disease progression in ASP. Toll‐like receptors (TLRs) are expressed on cells of the innate immune system, including glia, and TLR4 dysregulation may play a role in ASP pathogenesis. In this study we aimed to define the involvement of TLR4 in microglial and astroglial activation induced by different forms of α‐synuclein (full length soluble, fibrillized, and C‐terminally truncated). Purified primary wild type (TLR4 +/+ ) and TLR4 deficient (TLR4 −/− ) murine microglial and astroglial cell cultures were treated with recombinant α‐synuclein and phagocytic activity, NFκB nuclear translocation, cytokine release, and reactive oxygen species (ROS) production were measured. We show that TLR4 mediates α‐synuclein‐induced microglial phagocytic activity, pro‐inflammatory cytokine release, and ROS production. TLR4 −/− astroglia present a suppressed pro‐inflammatory response and decreased ROS production triggered by α‐synuclein treatment. However, the uptake of α‐synuclein by primary astroglia is not dependent on TLR4 expression. Our results indicate the C‐terminally truncated form as the most potent inductor of TLR4‐dependent glial activation. The current findings suggest that TLR4 plays a modulatory role on glial pro‐inflammatory responses and ROS production triggered by α‐synuclein. In contrast to microglia, the uptake of alpha‐synuclein by astroglia is not dependent on TLR4. Our data provide novel insights into the mechanisms of α‐synuclein‐induced microglial and astroglial activation which may have an impact on understanding the pathogenesis of ASP. © 2012 Wiley Periodicals, Inc.

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References 63

1. 1.

   Akira S. Toll-like receptors and innate immunity. Adv Immunol. 2001;78:1-56.

   Not in our library10.1016/s0065-2776(01)78001-7

2. 2.
   Genetic Variants of the α-Synuclein Gene SNCA Are Associated with Multiple System Atrophy · 2009

   No notice
3. 3.

   Alfonso-Loeches S, Pascual-Lucas M, Blanco AM, Sanchez-Vera I, Guerri C. Pivotal role of TLR4 receptors in alcohol-induced neuroinflammation and brain damage. J Neurosci. 2010;30:8285-8295.

   Not in our library10.1523/jneurosci.0976-10.2010

4. 4.

   Alvarez-Erviti L, Couch Y, Richardson J, Cooper JM, Wood MJ. Alpha-synuclein release by neurons activates the inflammatory response in a microglial cell line. Neurosci Res. 2011;69:337-342.

   Not in our library10.1016/j.neures.2010.12.020

5. 5.

   Apetri MM, Maiti NC, Zagorski MG, Carey PR, Anderson VE. Secondary structure of alpha-synuclein oligomers: Characterization by raman and atomic force microscopy. J Mol Biol. 2006;355:63-71.

   Not in our library10.1016/j.jmb.2005.10.071

6. 6.

   Austin SA, Floden AM, Murphy EJ, Combs CK. Alpha-synuclein expression modulates microglial activation phenotype. J Neurosci. 2006;26:10558-10563.

   Not in our library10.1523/jneurosci.1799-06.2006

7. 7.

   Baba M, Nakajo S, Tu PH, Tomita T, Nakaya K, Lee VM, Trojanowski JQ, Iwatsubo T. Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson's disease and dementia with Lewy bodies. Am J Pathol. 1998;152:879-884.

   Not in our library

8. 8.

   Bolder SG, Sagis LM, Venema P, van der Linden E. Thioflavin T and birefringence assays to determine the conversion of proteins into fibrils. Langmuir. 2007;23:4144-4147.

   Not in our library10.1021/la063048k

9. 9.

   Bowman CC, Rasley A, Tranguch SL, Marriott I. Cultured astrocytes express toll-like receptors for bacterial products. Glia. 2003;43:281-291.

   Not in our library10.1002/glia.10256

10. 10.

    Bsibsi M, Ravid R, Gveric D, van Noort JM. Broad expression of Toll-like receptors in the human central nervous system. J Neuropathol Exp Neurol. 2002;61:1013-1021.

    Not in our library10.1093/jnen/61.11.1013

11. 11.

    Carpentier PA, Begolka WS, Olson JK, Elhofy A, Karpus WJ, Miller SD. Differential activation of astrocytes by innate and adaptive immune stimuli. Glia. 2005;49:360-374.

    Not in our library10.1002/glia.20117

12. 12.

    Cole NB, Murphy DD, Grider T, Rueter S, Brasaemle D, Nussbaum RL. Lipid droplet binding and oligomerization properties of the Parkinson's disease protein alpha-synuclein. J Biol Chem. 2002;277:6344-6352.

    Not in our library10.1074/jbc.m108414200

13. 13.

    Conway KA, Harper JD, Lansbury PT. Fibrils formed in vitro from alpha-synuclein and two mutant forms linked to Parkinson's disease are typical amyloid. Biochemistry. 2000;39:2552-2563.

    Not in our library10.1021/bi991447r

14. 14.

    El-Hage N, Podhaizer EM, Sturgill J, Hauser KF. Toll-like receptor expression and activation in astroglia: Differential regulation by HIV-1 Tat, gp120, and morphine. Immunol Invest. 2011;40:498-522.

    Not in our library10.3109/08820139.2011.561904

15. 15.

    Faissner A, Pyka M, Geissler M, Sobik T, Frischknecht R, Gundelfinger ED, Seidenbecher C. Contributions of astrocytes to synapse formation and maturation—Potential functions of the perisynaptic extracellular matrix. Brain Res Rev. 2010;63:26-38.

    Not in our library10.1016/j.brainresrev.2010.01.001

16. 16.

    Fellner L, Jellinger KA, Wenning GK, Stefanova N. Glial dysfunction in the pathogenesis of alpha-synucleinopathies: Emerging concepts. Acta Neuropathol. 2011;121:675-693.

    Not in our library10.1007/s00401-011-0833-z

17. 17.

    Gai WP, Power JH, Blumbergs PC, Culvenor JG, Jensen PH. Alpha-synuclein immunoisolation of glial inclusions from multiple system atrophy brain tissue reveals multiprotein components. J Neurochem. 1999;73:2093-2100.

    Not in our library

18. 18.

    Gao HM, Hong JS, Zhang W, Liu B. Synergistic dopaminergic neurotoxicity of the pesticide rotenone and inflammogen lipopolysaccharide: Relevance to the etiology of Parkinson's disease. J Neurosci. 2003;23:1228-1236.

    Not in our library10.1523/jneurosci.23-04-01228.2003

19. 19.

    Gasser T. Molecular pathogenesis of Parkinson disease: Insights from genetic studies. Expert Rev Mol Med. 2009;11:e22.

    Not in our library10.1017/s1462399409001148

20. 20.

    Gerhard A, Banati RB, Goerres GB, Cagnin A, Myers R, Gunn RN, Turkheimer F, Good CD, Mathias CJ, Quinn N, Schwarz J, Brooks DJ. [11C](R)-PK11195 PET imaging of microglial activation in multiple system atrophy. Neurology. 2003;61:686-689.

    Not in our library10.1212/01.wnl.0000078192.95645.e6

21. 21.

    Gerhard A, Pavese N, Hotton G, Turkheimer F, Es M, Hammers A, Eggert K, Oertel W, Banati RB, Brooks DJ. In vivo imaging of microglial activation with [11C](R)-PK11195 PET in idiopathic Parkinson's disease. Neurobiol Dis. 2006;21:404-412.

    Not in our library10.1016/j.nbd.2005.08.002

22. 22.

    Glezer I, Simard AR, Rivest S. Neuroprotective role of the innate immune system by microglia. Neuroscience. 2007;147:867-883.

    Not in our library10.1016/j.neuroscience.2007.02.055

23. 23.

    Gorina R, Font-Nieves M, Marquez-Kisinousky L, Santalucia T, Planas AM. Astrocyte TLR4 activation induces a proinflammatory environment through the interplay between MyD88-dependent NFkappaB signaling, MAPK, and Jak1/Stat1 pathways. Glia. 2011;59:242-255.

    Not in our library10.1002/glia.21094

24. 24.

    Halliday GM, Stevens CH. Glia: Initiators and progressors of pathology in Parkinson's disease. Mov Disord. 2011;26:6-17.

    Not in our library10.1002/mds.23455

25. 25.

    Hirsch EC, Hunot S, Hartmann A. Neuroinflammatory processes in Parkinson's disease. Parkinsonism Relat Disord. 2005;11(Suppl 1):S9-S15.

    Not in our library10.1016/j.parkreldis.2004.10.013

26. 26.

    Imamura K, Hishikawa N, Ono K, Suzuki H, Sawada M, Nagatsu T, Yoshida M, Hashizume Y. Cytokine production of activated microglia and decrease in neurotrophic factors of neurons in the hippocampus of Lewy body disease brains. Acta Neuropathol. 2005;109:141-150.

    Not in our library10.1007/s00401-004-0919-y

27. 27.

    Kielian T. Toll-like receptors in central nervous system glial inflammation and homeostasis. J Neurosci Res. 2006;83:711-730.

    Not in our library10.1002/jnr.20767

28. 28.

    Klegeris A, Giasson BI, Zhang H, Maguire J, Pelech S, McGeer PL. Alpha-synuclein and its disease-causing mutants induce ICAM-1 and IL-6 in human astrocytes and astrocytoma cells. Faseb J. 2006;20:2000-2008.

    Not in our library10.1096/fj.06-6183com

29. 29.

    Klegeris A, Pelech S, Giasson BI, Maguire J, Zhang H, McGeer EG, McGeer PL. Alpha-synuclein activates stress signaling protein kinases in THP-1 cells and microglia. Neurobiol Aging. 2008;29:739-752.

    Not in our library10.1016/j.neurobiolaging.2006.11.013

30. 30.

    Koehler RC, Roman RJ, Harder DR. Astrocytes and the regulation of cerebral blood flow. Trends Neurosci. 2009;32:160-169.

    Not in our library10.1016/j.tins.2008.11.005

31. 31.

    Lee EJ, Woo MS, Moon PG, Baek MC, Choi IY, Kim WK, Junn E, Kim HS. Alpha-synuclein activates microglia by inducing the expressions of matrix metalloproteinases and the subsequent activation of protease-activated receptor-1. J Immunol. 2010a;185:615-623.

    Not in our library10.4049/jimmunol.0903480

32. 32.
    Alpha‐Synuclein Stimulation of Astrocytes: Potential Role for Neuroinflammation and Neuroprotection · 2010

    No notice
33. 33.

    Lee HJ, Suk JE, Bae EJ, Lee SJ. Clearance and deposition of extracellular alpha-synuclein aggregates in microglia. Biochem Biophys Res Commun. 2008;372:423-428.

    Not in our library10.1016/j.bbrc.2008.05.045

34. 34.

    Lee HJ, Suk JE, Patrick C, Bae EJ, Cho JH, Rho S, Hwang D, Masliah E, Lee SJ. Direct transfer of alpha-synuclein from neuron to astroglia causes inflammatory responses in synucleinopathies. J Biol Chem. 2010c;285:9262-9272.

    Not in our library10.1074/jbc.m109.081125

35. 35.

    Lehnardt S. Innate immunity and neuroinflammation in the CNS: the role of microglia in Toll-like receptor-mediated neuronal injury. Glia. 2010;58:253-263.

    Not in our library10.1002/glia.20928

36. 36.

    Letiembre M, Liu Y, Walter S, Hao W, Pfander T, Wrede A, Schulz-Schaeffer W, Fassbender K. Screening of innate immune receptors in neurodegenerative diseases: A similar pattern. Neurobiol Aging. 2009;30:759-768.

    Not in our library10.1016/j.neurobiolaging.2007.08.018

37. 37.

    Liu J, Zhou Y, Wang Y, Fong H, Murray TM, Zhang J. Identification of proteins involved in microglial endocytosis of alpha-synuclein. J Proteome Res. 2007;6:3614-3627.

    Not in our library10.1021/pr0701512

38. 38.

    Nimmerjahn A, Kirchhoff F, Helmchen F. Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science. 2005;308:1314-1318.

    Not in our library10.1126/science.1110647

39. 39.

    Okun E, Griffioen KJ, Lathia JD, Tang SC, Mattson MP, Arumugam TV. Toll-like receptors in neurodegeneration. Brain Res Rev. 2009;59:278-292.

    Not in our library10.1016/j.brainresrev.2008.09.001

40. 40.

    Ozawa T, Paviour D, Quinn NP, Josephs KA, Sangha H, Kilford L, Healy DG, Wood NW, Lees AJ, Holton JL, Revesz T. The spectrum of pathological involvement of the striatonigral and olivopontocerebellar systems in multiple system atrophy: Clinicopathological correlations. Brain. 2004;127:2657-2671.

    Not in our library10.1093/brain/awh303

41. 41.

    Park JY, Paik SR, Jou I, Park SM. Microglial phagocytosis is enhanced by monomeric alpha-synuclein, not aggregated alpha-synuclein: implications for Parkinson's disease. Glia. 2008;56:1215-1223.

    Not in our library10.1002/glia.20691

42. 42.

    Prasad K, Beach TG, Hedreen J, Richfield EK. Critical role of truncated alpha-synuclein and aggregates in Parkinson's disease and incidental lewy body disease. Brain Pathol. 2012.

    Not in our library10.1111/j.1750-3639.2012.00597.x

43. 43.

    Prinz M, Priller J, Sisodia SS, Ransohoff RM. Heterogeneity of CNS myeloid cells and their roles in neurodegeneration. Nat Neurosci. 2011;14:1227-1235.

    Not in our library10.1038/nn.2923

44. 44.

    Rappold PM, Tieu K. Astrocytes and therapeutics for Parkinson's disease. Neurotherapeutics. 2010;7:413-423.

    Not in our library10.1016/j.nurt.2010.07.001

45. 45.

    Reed-Geaghan EG, Savage JC, Hise AG, Landreth GE. CD14 and toll-like receptors 2 and 4 are required for fibrillar A{beta}-stimulated microglial activation. J Neurosci. 2009;29:11982-11992.

    Not in our library10.1523/jneurosci.3158-09.2009

46. 46.

    Reynolds AD, Kadiu I, Garg SK, Glanzer JG, Nordgren T, Ciborowski P, Banerjee R, Gendelman HE. Nitrated alpha-synuclein and microglial neuroregulatory activities. J Neuroimmune Pharmacol. 2008;3:59-74.

    Not in our library10.1007/s11481-008-9100-z

47. 47.

    Rojanathammanee L, Murphy EJ, Combs CK. Expression of mutant alpha-synuclein modulates microglial phenotype in vitro. J Neuroinflammation. 2011;8:44.

    Not in our library10.1186/1742-2094-8-44

48. 48.

    Roodveldt C, Labrador-Garrido A, Gonzalez-Rey E, Fernandez-Montesinos R, Caro M, Lachaud CC, Waudby CA, Delgado M, Dobson CM, Pozo D. Glial innate immunity generated by non-aggregated alpha-synuclein in mouse: Differences between wild-type and Parkinson's disease-linked mutants. PLoS One. 2010;5:e13481.

    Not in our library10.1371/journal.pone.0013481

49. 49.

    Saijo K, Winner B, Carson CT, Collier JG, Boyer L, Rosenfeld MG, Gage FH, Glass CK. A Nurr1/CoREST pathway in microglia and astrocytes protects dopaminergic neurons from inflammation-induced death. Cell. 2009;137:47-59.

    Not in our library10.1016/j.cell.2009.01.038

50. 50.

    Scholz SW, Houlden H, Schulte C, Sharma M, Li A, Berg D, Melchers A, Paudel R, Gibbs JR, Simon-Sanchez J, Paisan-Ruiz C, Bras J, Ding J, Chen H, Traynor BJ, Arepalli S, Zonozi RR, Revesz T, Holton J, Wood N, Lees A, Oertel W, Wullner U, Goldwurm S, Pellecchia MT, Illig T, Riess O, Fernandez HH, Rodriguez RL, Okun MS, Poewe W, Wenning GK, Hardy JA, Singleton AB, Del Sorbo F, Schneider S, Bhatia KP, Gasser T. SNCA variants are associated with increased risk for multiple system atrophy. Ann Neurol. 2009;65:610-614.

    Not in our library10.1002/ana.21685

51. 51.

    Shults CW, Rockenstein E, Crews L, Adame A, Mante M, Larrea G, Hashimoto M, Song D, Iwatsubo T, Tsuboi K, Masliah E. Neurological and neurodegenerative alterations in a transgenic mouse model expressing human alpha-synuclein under oligodendrocyte promoter: Implications for multiple system atrophy. J Neurosci. 2005;25:10689-10699.

    Not in our library10.1523/jneurosci.3527-05.2005

52. 52.

    Simard M, Nedergaard M. The neurobiology of glia in the context of water and ion homeostasis. Neuroscience. 2004;129:877-896.

    Not in our library10.1016/j.neuroscience.2004.09.053

53. 53.

    Stefanova N, Fellner L, Reindl M, Masliah E, Poewe W, Wenning GK. Toll-like receptor 4 promotes α-synuclein clearance and survival of nigral dopaminergic neurons. Am J Pathol. 2011;179:954-963.

    Not in our library10.1016/j.ajpath.2011.04.013

54. 54.

    Stefanova N, Klimaschewski L, Poewe W, Wenning GK, Reindl M. Glial cell death induced by overexpression of alpha-synuclein. J Neurosci Res. 2001;65:432-438.

    Not in our library10.1002/jnr.1171

55. 55.

    Stefanova N, Reindl M, Neumann M, Kahle PJ, Poewe W, Wenning GK. Microglial activation mediates neurodegeneration related to oligodendroglial alpha-synucleinopathy: Implications for multiple system atrophy. Mov Disord. 2007;22:2196-2203.

    Not in our library10.1002/mds.21671

56. 56.

    Stefanova N, Schanda K, Klimaschewski L, Poewe W, Wenning GK, Reindl M. Tumor necrosis factor-alpha-induced cell death in U373 cells overexpressing alpha-synuclein. J Neurosci Res. 2003;73:334-340.

    Not in our library10.1002/jnr.10662

57. 57.

    Stoll G, Jander S. The role of microglia and macrophages in the pathophysiology of the CNS. Prog Neurobiol. 1999;58:233-247.

    Not in our library10.1016/s0301-0082(98)00083-5

58. 58.

    Streit WJ. Microglia as neuroprotective, immunocompetent cells of the CNS. Glia. 2002;40:133-139.

    Not in our library10.1002/glia.10154

59. 59.

    Su X, Maguire-Zeiss KA, Giuliano R, Prifti L, Venkatesh K, Federoff HJ. Synuclein activates microglia in a model of Parkinson's disease. Neurobiol Aging. 2008;29:1690-1701.

    Not in our library10.1016/j.neurobiolaging.2007.04.006

60. 60.

    Wakabayashi K, Hayashi S, Yoshimoto M, Kudo H, Takahashi H. NACP/alpha-synuclein-positive filamentous inclusions in astrocytes and oligodendrocytes of Parkinson's disease brains. Acta Neuropathol. 2000;99:14-20.

    Not in our library10.1007/pl00007400

61. 61.

    Wilhelmsson U, Bushong EA, Price DL, Smarr BL, Phung V, Terada M, Ellisman MH, Pekny M. Redefining the concept of reactive astrocytes as cells that remain within their unique domains upon reaction to injury. Proc Natl Acad Sci USA. 2006;103:17513-17518.

    Not in our library10.1073/pnas.0602841103

62. 62.
    Abberant α-Synuclein Confers Toxicity to Neurons in Part through Inhibition of Chaperone-Mediated Autophagy · 2009

    No notice
63. 63.

    Zhang W, Wang T, Pei Z, Miller DS, Wu X, Block ML, Wilson B, Zhou Y, Hong JS, Zhang J. Aggregated alpha-synuclein activates microglia: A process leading to disease progression in Parkinson's disease. Faseb J. 2005;19:533-542.

    Not in our library10.1096/fj.04-2751com

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