In vivo RNAi-mediated alpha-synuclein silencing induces nigrostriatal degeneration.
Two small-interfering RNAs (siRNAs) targeting alpha-synuclein (alpha-syn) and three control siRNAs were cloned in an adeno-associated virus (AAV) vector and unilaterally injected into rat substantia nigra pars compacta (SNc). Reduction of alpha-syn resulted in a rapid (4 week) reduction in the number of tyrosine hydroxylase (TH) positive cells and striatal dopamine (DA) on the injected side. The level of neurodegeneration induced by the different siRNAs correlated with their ability to downregulate alpha-syn protein and mRNA in tissue culture and in vivo. Examination of various SNc neuronal markers indicated that neurodegeneration was due to cell loss and not just downregulation of DA synthesis. Reduction of alpha-syn also resulted in a pronounced amphetamine induced behavioral asymmetry consistent with the level of neurodegeneration. In contrast, none of the three control siRNAs, which targeted genes not normally expressed in SNc, showed evidence of neurodegeneration or behavioral asymmetry, even at longer survival times. Moreover, co-expression of both rat alpha-syn and alpha-syn siRNA partially reversed the neurodegenerative and behavioral effects of alpha-syn siRNA alone. Our data show that alpha-syn plays an important role in the rat SNc and suggest that both up- and downregulation of wild-type alpha-syn expression increase the risk of nigrostriatal pathology.
Gorbatyuk
Oleg S
OS
Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gene Therapy Center Cancer/Genetics Research Complex, Building 1376 1376 Mowry Road, Room CG-208 Gainesville, FL 32610, USA. olegor@ufl.edu
Li
Shoudong
S
Nash
Kevin
K
Gorbatyuk
Marina
M
Lewin
Alfred S
AS
Sullivan
Layla F
LF
Mandel
Ronald J
RJ
Chen
Weijun
W
Meyers
Craig
C
Manfredsson
Fredric P
FP
Muzyczka
Nicholas
N
D000818:Animals; D001921:Brain; D020868:Gene Silencing / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D034622:RNA Interference; D051381:Rats; D013378:Substantia Nigra / Q000378:metabolism* / Q000473:pathology*; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
D051844:alpha-Synuclein