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Stereotaxic Targeting of Alpha-Synuclein Pathology in Mouse Brain Using Preformed Fibrils

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Book cover Alpha-Synuclein

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1948))

Abstract

The accumulation of intraneuronal inclusions containing misfolded alpha-synuclein (aSyn) within the central nervous system (CNS) is a common feature found in several neurodegenerative disorders including Parkinson’s disease (PD). Emerging evidence indicates that aSyn amyloid fibrils, a configuration that is present within these characteristic inclusions, are capable of self-replicating by templating the conversion of endogenously expressed aSyn in neurons. Stereotaxic administration of synthetic α-synuclein preformed fibrils (PFFs) into the mouse brain has been shown to seed the formation of intracellular aSyn pathology reminiscent of Lewy body (LB) inclusions present in human PD and related synucleinopathies. Moreover, pathology can be targeted to specific CNS regions. This experimental approach provides a versatile platform for investigating PD-like LB pathology in vivo. We focus here on procedures for initiating aSyn inclusion formation at various regions of the mouse brain using computer-assisted motorized stereotaxic microinjection of aSyn PFFs and discuss appropriate strategies for controls and analysis.

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References

  1. Toledo JB, Gopal P, Raible K et al (2016) Pathological α-synuclein distribution in subjects with coincident Alzheimer’s and Lewy body pathology. Acta Neuropathol 131:393–409

    Article  CAS  Google Scholar 

  2. Jellinger KA (2004) Lewy body-related alpha-synucleinopathy in the aged human brain. J Neural Transm (Vienna) 111:1219–1235

    Article  CAS  Google Scholar 

  3. Polymeropoulos MH, Lavedan C, Leroy E et al (1997) Mutation in the alpha-synuclein gene identified in families with Parkinson’s disease. Science 276:2045–2047

    Article  CAS  Google Scholar 

  4. Singleton AB, Farrer M, Johnson J et al (2003) alpha-Synuclein locus triplication causes Parkinson’s disease. Science 302:841

    Article  CAS  Google Scholar 

  5. Zarranz JJ, Alegre J, Gómez-Esteban JC et al (2004) The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 55:164–173

    Article  CAS  Google Scholar 

  6. Lesage S, Anheim M, Letournel F et al (2013) G51D α-synuclein mutation causes a novel parkinsonian-pyramidal syndrome. Ann Neurol 73:459–471

    Article  CAS  Google Scholar 

  7. Halliday GM, Del Tredici K, and Braak H (2006) Critical appraisal of brain pathology staging related to presymptomatic and symptomatic cases of sporadic Parkinson’s disease. J Neural Transm Suppl 70:99–103

    Google Scholar 

  8. Dickson DW, Uchikado H, Fujishiro H et al (2010) Evidence in favor of Braak staging of Parkinson’s disease. Mov Disord 25(Suppl 1):S78–S82

    Article  Google Scholar 

  9. Spillantini MG, Crowther RA, Jakes R et al (1998) Filamentous alpha-synuclein inclusions link multiple system atrophy with Parkinson’s disease and dementia with Lewy bodies. Neurosci Lett 251:205–208

    Article  CAS  Google Scholar 

  10. Braak H, Del Tredici K, Rüb U et al (2003) Staging of brain pathology related to sporadic Parkinson’s disease. Neurobiol Aging 24:197–211

    Article  Google Scholar 

  11. Burke RE, Dauer WT, Vonsattel JPG (2008) A critical evaluation of the Braak staging scheme for Parkinson’s disease. Ann Neurol 64:485–491

    Article  Google Scholar 

  12. Braak H, Rüb U, Gai WP et al (2003) Idiopathic Parkinson’s disease: possible routes by which vulnerable neuronal types may be subject to neuroinvasion by an unknown pathogen. J Neural Transm (Vienna) 110:517–536

    Article  CAS  Google Scholar 

  13. Wakabayashi K, Takahashi H, Takeda S et al (1988) Parkinson’s disease: the presence of Lewy bodies in Auerbach’s and Meissner’s plexuses. Acta Neuropathol 76:217–221

    Article  CAS  Google Scholar 

  14. Duda JE, Shah U, Arnold SE et al (1999) The expression of alpha-, beta-, and gamma-synucleins in olfactory mucosa from patients with and without neurodegenerative diseases. Exp Neurol 160:515–522

    Article  CAS  Google Scholar 

  15. Kordower JH, Chu Y, Hauser RA et al (2008) Lewy body-like pathology in long-term embryonic nigral transplants in Parkinson’s disease. Nat Med 14:504–506

    Article  CAS  Google Scholar 

  16. Li J-Y, Englund E, Holton JL et al (2008) Lewy bodies in grafted neurons in subjects with Parkinson’s disease suggest host-to-graft disease propagation. Nat Med 14:501–503

    Article  CAS  Google Scholar 

  17. Baba M, Nakajo S, Tu PH et al (1998) Aggregation of alpha-synuclein in Lewy bodies of sporadic Parkinson’s disease and dementia with Lewy bodies. Am J Pathol 152:879–884

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Spillantini MG, Crowther RA, Jakes R et al (1998) alpha-Synuclein in filamentous inclusions of Lewy bodies from Parkinson’s disease and dementia with Lewy bodies. Proc Natl Acad Sci U S A 95:6469–6473

    Article  CAS  Google Scholar 

  19. Fauvet B, Mbefo MK, Fares M-B et al (2012) α-Synuclein in central nervous system and from erythrocytes, mammalian cells, and Escherichia coli exists predominantly as disordered monomer. J Biol Chem 287:15345–15364

    Article  CAS  Google Scholar 

  20. Crowther RA, Jakes R, Spillantini MG et al (1998) Synthetic filaments assembled from C-terminally truncated alpha-synuclein. FEBS Lett 436:309–312

    Article  CAS  Google Scholar 

  21. Conway KA, Harper JD, Lansbury PT (1998) Accelerated in vitro fibril formation by a mutant alpha-synuclein linked to early-onset Parkinson disease. Nat Med 4:1318–1320

    Article  CAS  Google Scholar 

  22. Come JH, Fraser PE, Lansbury PT (1993) A kinetic model for amyloid formation in the prion diseases: importance of seeding. Proc Natl Acad Sci U S A 90:5959–5963

    Article  CAS  Google Scholar 

  23. Luk KC, Song C, O’Brien P et al (2009) Exogenous alpha-synuclein fibrils seed the formation of Lewy body-like intracellular inclusions in cultured cells. Proc Natl Acad Sci U S A 106:20051–20056

    Article  CAS  Google Scholar 

  24. Volpicelli-Daley LA, Luk KC, Patel TP et al (2011) Exogenous α-synuclein fibrils induce Lewy body pathology leading to synaptic dysfunction and neuron death. Neuron 72:57–71

    Article  CAS  Google Scholar 

  25. Luk KC, Kehm VM, Zhang B et al (2012) Intracerebral inoculation of pathological α-synuclein initiates a rapidly progressive neurodegenerative α-synucleinopathy in mice. J Exp Med 209:975–986

    Article  CAS  Google Scholar 

  26. Luk KC, Kehm V, Carroll J et al (2012) Pathological -synuclein transmission initiates Parkinson-like neurodegeneration in nontransgenic mice. Science 338:949–953

    Article  CAS  Google Scholar 

  27. Recasens A, Dehay B, Bové J et al (2014) Lewy body extracts from Parkinson disease brains trigger α-synuclein pathology and neurodegeneration in mice and monkeys: LB-induced pathology. Ann Neurol 75:351–362

    Article  CAS  Google Scholar 

  28. Masuda-Suzukake M, Nonaka T, Hosokawa M et al (2013) Prion-like spreading of pathological α-synuclein in brain. Brain J Neurol 136:1128–1138

    Article  Google Scholar 

  29. Bernis ME, Babila JT, Breid S et al (2015) Prion-like propagation of human brain-derived alpha-synuclein in transgenic mice expressing human wild-type alpha-synuclein. Acta Neuropathol Commun 3:75

    Article  Google Scholar 

  30. Watts JC, Giles K, Oehler A et al (2013) Transmission of multiple system atrophy prions to transgenic mice. Proc Natl Acad Sci U S A 110:19555–19560

    Article  CAS  Google Scholar 

  31. Bousset L, Pieri L, Ruiz-Arlandis G et al (2013) Structural and functional characterization of two alpha-synuclein strains. Nat Commun 4:2575

    Article  Google Scholar 

  32. Guo JL, Covell DJ, Daniels JP et al (2013) Distinct α-synuclein strains differentially promote tau inclusions in neurons. Cell 154:103–117

    Article  CAS  Google Scholar 

  33. Paumier KL, Luk KC, Manfredsson FP et al (2015) Intrastriatal injection of pre-formed mouse α-synuclein fibrils into rats triggers α-synuclein pathology and bilateral nigrostriatal degeneration. Neurobiol Dis 82:185–199

    Article  CAS  Google Scholar 

  34. Shimozawa A, Ono M, Takahara D et al (2017) Propagation of pathological α-synuclein in marmoset brain. Acta Neuropathol Commun 5:12

    Article  Google Scholar 

  35. Koprich JB, Kalia LV, Brotchie JM (2017) Animal models of α-synucleinopathy for Parkinson disease drug development. Nat Rev Neurosci 18:515–529

    Article  CAS  Google Scholar 

  36. Luna E, Luk KC (2015) Bent out of shape: α-Synuclein misfolding and the convergence of pathogenic pathways in Parkinson’s disease. FEBS Lett 589:3749–3759

    Article  CAS  Google Scholar 

  37. Dehay B, Vila M, Bezard E et al (2016) Alpha-synuclein propagation: new insights from animal models. Mov Disord 31:161–168

    Article  Google Scholar 

  38. Sacino AN, Brooks M, Thomas MA et al (2014) Intramuscular injection of -synuclein induces CNS -synuclein pathology and a rapid-onset motor phenotype in transgenic mice. Proc Natl Acad Sci 111:10732–10737

    Article  CAS  Google Scholar 

  39. Holmqvist S, Chutna O, Bousset L et al (2014) Direct evidence of Parkinson pathology spread from the gastrointestinal tract to the brain in rats. Acta Neuropathol 128:805–820

    Article  Google Scholar 

  40. Manfredsson FP, Luk KC, Benskey MJ et al (2018) Induction of alpha-synuclein pathology in the enteric nervous system of the rat and non-human primate results in gastrointestinal dysmotility and transient CNS pathology. Neurobiol Dis 112:106–118

    Article  CAS  Google Scholar 

  41. Mason DM, Nouraei N, Pant DB et al (2016) Transmission of α-synucleinopathy from olfactory structures deep into the temporal lobe. Mol Neurodegener 11:49

    Article  Google Scholar 

  42. Nouraei N, Mason DM, Miner KM et al (2018) Critical appraisal of pathology transmission in the α-synuclein fibril model of Lewy body disorders. Exp Neurol 299:172–196

    Article  CAS  Google Scholar 

  43. Rey NL, Steiner JA, Maroof N et al (2016) Widespread transneuronal propagation of α-synucleinopathy triggered in olfactory bulb mimics prodromal Parkinson’s disease. J Exp Med 213:1759–1778

    Article  CAS  Google Scholar 

  44. Osterberg VR, Spinelli KJ, Weston LJ et al (2015) Progressive aggregation of alpha-synuclein and selective degeneration of Lewy inclusion-bearing neurons in a mouse model of Parkinsonism. Cell Rep 10:1252–1260

    Article  CAS  Google Scholar 

  45. Masuda-Suzukake M, Nonaka T, Hosokawa M et al (2014) Pathological alpha-synuclein propagates through neural networks. Acta Neuropathol Commun 2:88

    Article  Google Scholar 

  46. Polinski NK, Volpicelli-Daley LA, Sortwell CE et al (2018) Best practices for generating and using alpha-synuclein pre-formed fibrils to model Parkinson’s disease in rodents. J Parkinsons Dis 8(2):303–322

    Article  Google Scholar 

  47. Bousset L, Brundin P, Böckmann A et al (2016) An efficient procedure for removal and inactivation of alpha-synuclein assemblies from laboratory materials. J Parkinsons Dis 6:143–151

    Article  CAS  Google Scholar 

  48. Abdelmotilib H, Maltbie T, Delic V et al (2017) α-Synuclein fibril-induced inclusion spread in rats and mice correlates with dopaminergic Neurodegeneration. Neurobiol Dis 105:84–98

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported in part by NIH grants NS088322 and NS053488.

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Authors and Affiliations

  1. Center for Neurodegenerative Disease Research, Institute on Aging, Department of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Bin Zhang, Victoria Kehm, Ron Gathagan, Susan N. Leight, John Q. Trojanowski, Virginia M.-Y. Lee & Kelvin C. Luk

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  1. Bin Zhang
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  2. Victoria Kehm
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  3. Ron Gathagan
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  4. Susan N. Leight
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  5. John Q. Trojanowski
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  6. Virginia M.-Y. Lee
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  7. Kelvin C. Luk
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Corresponding author

Correspondence to Kelvin C. Luk .

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Editors and Affiliations

  1. Dementia Research Institute, University College London, London, UK

    Tim Bartels

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Zhang, B. et al. (2019). Stereotaxic Targeting of Alpha-Synuclein Pathology in Mouse Brain Using Preformed Fibrils. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_5

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  • DOI: https://doi.org/10.1007/978-1-4939-9124-2_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9123-5

  • Online ISBN: 978-1-4939-9124-2

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