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RNA interference of LRRK2–microarray expression analysis of a Parkinson’s disease key player

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Abstract

The protein leucine-rich repeat kinase 2 (LRRK2) is a key player in the pathogenesis of Parkinson’s disease (PD). Mutations in the LRRK2 gene account for up to 10% of all autosomal dominant forms of familiar and for approximately 1–3% of sporadic PD patients. Although the LRRK2 protein has many functional domains like a leucine-rich repeat domain, a Roc-GTPase domain, a kinase domain of the tyrosine kinase-like subfamily and multiple protein interaction domains (armadillo, ankyrin, WD40), the exact biological role of LRRK2 in the human brain is elusive. To gain more insight into the biological function of this protein, we monitored the changes in the expression profiles of SH-SY5Y cells, a dopaminergic neuroblastoma cell line, induced by a depletion of LRRK2 levels by RNA interference (RNAi) with Affymetrix U133 Plus 2.0 microarrays. A total of 187 genes were differentially regulated by at least a 1.5-fold change with 94 transcripts being upregulated and 93 transcripts being downregulated compared to scrambled control siRNA transfected cells. Key players of the interaction networks were independently verified by qRT-PCR. The differentially expressed gene products are involved in axonal guidance, nervous system development, cell cycle, cell growth, cell differentiation, cell communication, MAPKKK cascade, and Ras protein signal transduction. Defined gene expression networks will now serve to look more closely for candidates affected by LRRK2 reduction and how they might be altered in other forms of familial or sporadic PD.

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Acknowledgement

We thank S. Biskup and P. Kahle for helpful discussion and reading the manuscript. This work was supported by IZKF (01KS 9602/4)/BMBF and an NGFN2 grant to OR. Experiments comply with the current laws in Germany.

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Authors and Affiliations

  1. Department of Medical Genetics, Microarray Facility, University of Tübingen, Calwerstrasse 7, 72076, Tübingen, Germany

    K. Häbig, M. Walter, S. Poths, O. Riess & M. Bonin

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  1. K. Häbig
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  2. M. Walter
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  3. S. Poths
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  4. O. Riess
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  5. M. Bonin
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Correspondence to M. Bonin.

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Supplementary material 1

Differentially expressed transcripts of the LRRK2 RNAi microarray experiment. (DOC 487 KB)

Supplementary material 2

Differentially expressed transcripts of the 16 presented GO-categories. (DOC 239 KB)

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Häbig, K., Walter, M., Poths, S. et al. RNA interference of LRRK2–microarray expression analysis of a Parkinson’s disease key player. Neurogenetics 9, 83–94 (2008). https://doi.org/10.1007/s10048-007-0114-0

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  • DOI: https://doi.org/10.1007/s10048-007-0114-0

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