Seeded aggregation and toxicity of {alpha}-synuclein and tau: cellular models of neurodegenerative diseases.
The deposition of amyloid-like filaments in the brain is the central event in the pathogenesis of neurodegenerative diseases. Here we report cellular models of intracytoplasmic inclusions of α-synuclein, generated by introducing nucleation seeds into SH-SY5Y cells with a transfection reagent. Upon introduction of preformed seeds into cells overexpressing α-synuclein, abundant, highly filamentous α-synuclein-positive inclusions, which are extensively phosphorylated and ubiquitinated and partially thioflavin-positive, were formed within the cells. SH-SY5Y cells that formed such inclusions underwent cell death, which was blocked by small molecular compounds that inhibit β-sheet formation. Similar seed-dependent aggregation was observed in cells expressing four-repeat Tau by introducing four-repeat Tau fibrils but not three-repeat Tau fibrils or α-synuclein fibrils. No aggregate formation was observed in cells overexpressing three-repeat Tau upon treatment with four-repeat Tau fibrils. Our cellular models thus provide evidence of nucleation-dependent and protein-specific polymerization of intracellular amyloid-like proteins in cultured cells.
Nonaka
Takashi
T
Department of Molecular Neurobiology, Tokyo Institute of Psychiatry, Tokyo 156-8585, USA. nonaka-tk@igakuken.or.jp
Watanabe
Sayuri T
ST
Iwatsubo
Takeshi
T
Hasegawa
Masato
M
D016923:Cell Death; D045744:Cell Line, Tumor; D006801:Humans; D002479:Inclusion Bodies / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D019636:Neurodegenerative Diseases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism*
C054369:MAPT protein, human; D051844:alpha-Synuclein; D016875:tau Proteins
Proc Natl Acad Sci U S A. 2009 Aug 4;106(31):13010-5
19651612
J Biol Chem. 1999 Jul 9;274(28):19509-12
10391881
N Engl J Med. 2001 May 17;344(20):1516-26
11357156
Biochemistry. 2009 Aug 25;48(33):8014-22
19630439
Science. 2001 May 25;292(5521):1552-5
11375494
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11183-6
7972031
Curr Opin Neurobiol. 1998 Oct;8(5):619-32
9811617
EMBO J. 1998 May 15;17(10):2759-66
9582269
Brain Res Mol Brain Res. 1997 Feb;44(1):105-12
9030704
Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4897-902
10781096
Nat Cell Biol. 2002 Feb;4(2):160-4
11813001
Trends Biochem Sci. 1999 Feb;24(2):58-63
10098399
Am J Pathol. 2000 Jul;157(1):277-86
10880397
Annu Rev Biochem. 1997;66:385-407
9242912
Trends Biochem Sci. 2006 Mar;31(3):150-5
16473510
Proc Natl Acad Sci U S A. 2003 Jul 22;100(15):9034-8
12853572
Acta Neuropathol. 1991;82(4):239-59
1759558
J Biol Chem. 2007 Jul 13;282(28):20309-18
17526496
Nat Cell Biol. 2009 Jul;11(7):909-13
19503072
Neuron. 2005 Jun 16;46(6):857-68
15953415
Proc Natl Acad Sci U S A. 1987 Oct;84(19):6953-7
3477820
Proc Natl Acad Sci U S A. 1998 May 26;95(11):6469-73
9600990
J Mol Biol. 1995 Sep 29;252(4):412-22
7563061
Science. 2001 Nov 9;294(5545):1346-9
11701929
Proc Natl Acad Sci U S A. 2009 Nov 24;106(47):20051-6
19892735
J Histochem Cytochem. 1968 Nov;16(11):673-7
5723775
Cell. 2005 Apr 8;121(1):49-62
15820678
Cell. 1993 Jun 18;73(6):1055-8
8513491
Biochemistry. 2005 Jan 11;44(1):361-8
15628878
J Biol Chem. 2005 Mar 4;280(9):7614-23
15611092
Annu Rev Biochem. 2006;75:333-66
16756495
Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1977-81
8446617
FEBS Lett. 1994 May 23;345(1):27-32
8194594
Brain Res. 1991 Feb 8;541(1):163-6
2029618
Hum Mol Genet. 2009 Sep 15;18(18):3353-64
19515851
J Biol Chem. 2001 Jan 26;276(4):2380-6
11060312
Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15712-7
9861035
J Biol Chem. 2002 Dec 13;277(50):49071-6
12377775
J Biol Chem. 2006 Nov 24;281(47):36180-6
17012232
Biochemistry. 2006 May 16;45(19):6085-94
16681381
J Biol Chem. 2009 Mar 20;284(12):7940-50
19164293
J Biol Chem. 2001 Jul 20;276(29):27441-8
11316809