Different species of alpha-synuclein oligomers induce calcium influx and seeding.
Aggregation of alpha-synuclein (alpha-syn) has been linked to the pathogenesis of Parkinson's disease (PD) and other neurodegenerative diseases. Increasing evidence suggests that prefibrillar oligomers and protofibrils, rather than mature fibrils of alpha-syn, are the pathogenic species in PD. Despite extensive effort on studying oligomerization of alpha-syn, no studies have compared different oligomer species directly on a single-particle level and investigated their biological effects on cells. In this study, we applied a novel highly sensitive single molecule detection system that allowed a direct comparison of different oligomer types. Furthermore, we studied biological effects of different oligomer types on cells. For this purpose, we developed new oligomerization protocols, that enabled the use of these different oligomers in cell culture. We found that all of our three aggregation protocols resulted in heterogeneous populations of oligomers. Some types of oligomers induced cell death via disruption of cellular ion homeostasis by a presumably pore-forming mechanism. Other oligomer types could directly enter the cell resulting in increased alpha-syn aggregation. Based on our results, we propose that under various physiological conditions, heterogeneous populations of oligomeric forms will coexist in an equilibrium. These different oligomer types lead directly or indirectly to cell damage. Our data indicate that inhibition of early alpha-syn aggregation events would consequently prevent all alpha-syn oligomer related toxicities. This has important implications for the development of disease-modifying drugs for the treatment of PD and other synucleinopathies.
Danzer
Karin M
KM
Central Nervous System Research, Boehringer Ingelheim Pharma, 88397 Biberach, Germany. Karin.Danzer@bc.boehringer-ingelheim.com
Haasen
Dorothea
D
Karow
Anne R
AR
Moussaud
Simon
S
Habeck
Matthias
M
Giese
Armin
A
Kretzschmar
Hans
H
Hengerer
Bastian
B
Kostka
Marcus
M
D000818:Animals; D001704:Biopolymers; D001921:Brain / Q000166:cytology; D002118:Calcium / Q000378:metabolism*; D053148:Caspase 3; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods; D009154:Mutation / Q000502:physiology; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects*; D010446:Peptide Fragments / Q000737:chemistry; D011994:Recombinant Proteins; D014162:Transfection / Q000379:methods; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*
D001704:Biopolymers; D010446:Peptide Fragments; D011994:Recombinant Proteins; D051844:alpha-Synuclein; D053148:Caspase 3; D002118:Calcium
J Biol Chem. 1999 Jul 9;274(28):19509-12
10391881
Proc Natl Acad Sci U S A. 1999 Nov 23;96(24):13756-61
10570145
Nat Struct Biol. 2000 Jan;7(1):48-53
10625427
Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):571-6
10639120
Cell. 2000 Jan 21;100(2):265-76
10660049
Science. 2000 Feb 18;287(5456):1265-9
10678833
Biophys J. 2000 Apr;78(4):1703-13
10733953
J Biol Chem. 2000 Jul 14;275(28):21500-7
10764738
J Biol Chem. 2000 May 12;275(19):14077-83
10799482
Nat Cell Biol. 2000 Jul;2(7):E115-9
10878819
Biochemistry. 2000 Sep 5;39(35):10619-26
10978144
Biochemistry. 2001 Jul 3;40(26):7812-9
11425308
Nat Rev Neurosci. 2001 Jul;2(7):492-501
11433374
J Biol Chem. 2001 Nov 23;276(47):44284-96
11553618
J Biol Chem. 2002 Jan 4;277(1):671-8
11679584
J Mol Neurosci. 2001 Oct;17(2):205-24
11816794
Science. 2002 Feb 1;295(5556):865-8
11823645
Biochemistry. 2002 Apr 9;41(14):4595-602
11926821
Nature. 2002 Apr 4;416(6880):507-11
11932737
Nature. 2002 Jul 18;418(6895):291
12124613
J Mol Biol. 2002 Sep 13;322(2):383-93
12217698
J Biol Chem. 2002 Dec 13;277(50):48984-92
12351643
J Mol Biol. 2002 Oct 4;322(5):1089-102
12367530
Neurobiol Aging. 2003 Mar-Apr;24(2):197-211
12498954
Biochemistry. 2003 Apr 8;42(13):3696-700
12667059
Science. 2003 Apr 18;300(5618):486-9
12702875
Neuron. 2003 Sep 11;39(6):889-909
12971891
Science. 2003 Oct 31;302(5646):841
14593171
Ann Neurol. 2004 Feb;55(2):164-73
14755719
J Membr Biol. 2004 Jan 1;197(1):33-48
15014916
J Biol Chem. 2004 May 21;279(21):21966-75
15028717
J Biol Chem. 2004 Jul 23;279(30):31374-82
15133040
J Mol Biol. 2004 Jun 25;340(1):127-39
15184027
Biochemistry. 2004 Jul 20;43(28):9021-8
15248759
J Mol Biol. 2004 Sep 17;342(3):877-87
15342243
J Biol Chem. 2004 Nov 5;279(45):46363-6
15385542
Lancet. 2004 Sep 25-Oct 1;364(9440):1167-9
15451224
Neurotox Res. 2005;7(1-2):59-67
15639798
J Biol Chem. 2005 Apr 29;280(17):17294-300
15722360
Biochem Biophys Res Commun. 2005 Apr 22;329(4):1200-7
15766554
J Neurosci. 2005 Jun 22;25(25):6016-24
15976091
Methods Mol Biol. 2005;299:19-33
15980593
J Mol Biol. 2006 Jan 6;355(1):63-71
16303137
Comb Chem High Throughput Screen. 2006 Jan;9(1):37-47
16454685
FASEB J. 2006 Mar;20(3):419-25
16507759
J Neurochem. 2006 Apr;97(1):265-79
16524375
Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4246-51
16537516
J Neurochem. 2006 May;97(4):1071-7
16606366
Comb Chem High Throughput Screen. 2006 Jun;9(5):339-50
16787147
Q Rev Biophys. 2006 May;39(2):167-201
16978447
Methods Enzymol. 2006;413:326-44
17046404
Neuroreport. 2006 Dec 18;17(18):1883-6
17179863
FEBS J. 2007 Apr;274(7):1862-77
17381514
Brain Res. 1995 Mar 13;674(1):133-6
7773681
Ann N Y Acad Sci. 1996 Jan 17;777:368-73
8624114
Science. 1997 Jun 27;276(5321):2045-7
9197268
J Neurochem. 1997 Oct;69(4):1746-52
9326304
Biochemistry. 1997 Oct 28;36(43):13298-304
9341221
Nat Genet. 1998 Feb;18(2):106-8
9462735