Linked Life Data

17131421

Source:http://linkedlifedata.com/resource/pubmed/id/17131421

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2007-1-22
pubmed:abstractText
The critical observation in the pathology of Parkinson's disease (PD) is that neurodegeneration is largely restricted to dopaminergic neurons that develop cytoplasmic inclusions called Lewy bodies. These aggregations contain the protein alpha-synuclein. Furthermore, it is becoming apparent that alpha-synuclein expression levels are a major factor in PD pathogenesis. Patients with additional copies of the alpha-synuclein gene develop PD with a severity proportional to levels of alpha-synuclein overexpression. Similarly, overexpression of alpha-synuclein in in vitro and in vivo models has been shown to be toxic. However, little is known about the effects of reducing alpha-synuclein expression in human neurons. To investigate this, we have developed a system in which levels of alpha-synuclein can be acutely suppressed by using RNA interference (RNAi) in a physiologically relevant human dopaminergic cellular model. By using small interfering RNA (siRNA) molecules targeted to endogenous alpha-synuclein, we achieved 80% protein knockdown. We show that alpha-synuclein knockdown has no effect on cellular survival either under normal growth conditions over 5 days or in the presence of the mitochondrial inhibitor rotenone. Knockdown does, however, confer resistance to the dopamine transporter (DAT)-dependent neurotoxin N-methyl-4-phenylpyridinium (MPP(+)). We then demonstrate for the first time that alpha-synuclein suppression decreases dopamine transport in human cells, reducing the maximal uptake velocity (V(max)) of dopamine and the surface density of its transporter by up to 50%. These results show that RNAi-mediated alpha-synuclein knockdown alters cellular dopamine homeostasis in human cells and may suggest a mechanism for the increased survival in the presence of MPP(+), a toxin used extensively to model Parkinson's disease.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0360-4012
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
85
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
351-63
pubmed:dateRevised
2010-12-3
pubmed:meshHeading
pubmed-meshheading:17131421-1-Methyl-4-phenylpyridinium, pubmed-meshheading:17131421-Blotting, Western, pubmed-meshheading:17131421-Cell Line, Tumor, pubmed-meshheading:17131421-Cell Survival, pubmed-meshheading:17131421-Dopamine, pubmed-meshheading:17131421-Dopamine Plasma Membrane Transport Proteins, pubmed-meshheading:17131421-Fluorescent Antibody Technique, pubmed-meshheading:17131421-Humans, pubmed-meshheading:17131421-Neuroblastoma, pubmed-meshheading:17131421-Neurons, pubmed-meshheading:17131421-Neurotoxins, pubmed-meshheading:17131421-Protein Transport, pubmed-meshheading:17131421-RNA Interference, pubmed-meshheading:17131421-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:17131421-Rotenone, pubmed-meshheading:17131421-Transfection, pubmed-meshheading:17131421-alpha-Synuclein
pubmed:year
2007
pubmed:articleTitle
RNA interference-mediated knockdown of alpha-synuclein protects human dopaminergic neuroblastoma cells from MPP(+) toxicity and reduces dopamine transport.
pubmed:affiliation
The Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, United Kingdom.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't