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Nucleic Acids Research Advance Access published online on March 10, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn084
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Elevated {alpha}-synuclein mRNA levels in individual UV-laser-microdissected dopaminergic substantia nigra neurons in idiopathic Parkinson's disease

Jan Gründemann1, Falk Schlaudraff1,2, Olga Haeckel1 and Birgit Liss1,2,*

1Molecular Neurobiology, Department of Physiology, Philipps-University Marburg, Deutschhausstrasse 2, 35037 Marburg and 2Molecular Neurophysiology, Institute for General Physiology, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany

*To whom correspondence should be addressed. Tel: +49 731 500 23115; Fax: +49 731 500 23242; Email: birgit.liss{at}uni-ulm.de

Received October 12, 2007. Revised February 8, 2008. Accepted February 11, 2008.

The presynaptic protein {alpha}-synuclein is involved in several neurodegenerative diseases, including Parkinson's disease (PD). In rare familial forms of PD, causal mutations (PARK1) as well as multiplications (PARK4) of the {alpha}-synuclein gene have been identified. In sporadic, idiopathic PD, abnormal accumulation and deposition of {alpha}-synuclein might also cause degeneration of dopaminergic midbrain neurons, the clinically most relevant neuronal population in PD. Thus, cell-specific quantification of {alpha}-synuclein expression-levels in dopaminergic neurons from idiopathic PD patients in comparison to controls would provide essential information about contributions of {alpha}-synuclein to the etiology of PD. However, a number of previous studies addressing this question at the tissue-level yielded varying results regarding {alpha}-synuclein expression. To increase specificity, we developed a cell-specific approach for mRNA quantification that also took into account the important issue of variable RNA integrities of the individual human postmortem brain samples. We demonstrate that PCR –amplicon size can confound quantitative gene-expression analysis, in particular of partly degraded RNA. By combining optimized UV-laser microdissection- and quantitative RT–PCR-techniques with suitable PCR assays, we detected significantly elevated {alpha}-synuclein mRNA levels in individual, surviving neuromelanin- and tyrosine hydroxylase-positive substantia nigra dopaminergic neurons from idiopathic PD brains compared to controls. These results strengthen the pathophysiologic role of transcriptional dysregulation of the {alpha}-synuclein gene in sporadic PD.

Present address: Jan Gründemann, Wolfson Institute for Biomedical Research, University College London, WC1E 6BT, London, UK


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