Abstract
Cell specificity of gene expression analysis is essential to avoid tissue sample related artifacts, in particular when the relative number of target cells present in the compared tissues varies dramatically, e.g., when comparing dopamine neurons in midbrain tissues from control subjects with those from Parkinson’s disease (PD) cases. Here, we describe a detailed protocol that combines contact-free UV-laser microdissection and quantitative PCR of reverse-transcribed RNA of individual neurons from postmortem human midbrain tissue from PD patients and unaffected controls. Among expression changes in a variety of dopamine neuron marker, maintenance, and cell-metabolism genes, we found that α-synuclein mRNA levels were significantly elevated in individual neuromelanin-positive dopamine midbrain neurons from PD brains when compared to those from matched controls.
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References
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Acknowledgments
We are particularly grateful to the brain donors and the support by the German BrainNet (GA28). We thank Leica Microsystems for providing the LMD6000 and Jochen Roeper for critical reading the manuscript. This work was supported by the BMBF (NGFN-Net), the DFG (SFB497), the Gemeinnützige Hertiestiftung, the Parkinson’s Disease society, and the Royal Society, UK. J.G. is supported by a PhD Studentship of the Wellcome Trust; B.L. is supported by the Alfried Krupp prize for young university teachers.
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Gründemann, J., Schlaudraff, F., Liss, B. (2011). UV-Laser Microdissection and mRNA Expression Analysis of Individual Neurons from Postmortem Parkinson’s Disease Brains. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 755. Humana Press. https://doi.org/10.1007/978-1-61779-163-5_30
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DOI: https://doi.org/10.1007/978-1-61779-163-5_30
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