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Normalization and expression changes in predefined sets of proteins using 2D gel electrophoresis : A proteomic study of L-DOPA induced dyskinesia in an animal model of Parkinson's disease using DIGE

Kultima, Kim (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Scholz, Birger (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Alm, Henrik (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
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Sköld, Karl (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Svensson, Marcus (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Crossman, Alan (author)
Bezard, Erwan (author)
Andrén, Per E. (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap
Lönnstedt, Ingrid (author)
Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Matematiska institutionen
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 (creator_code:org_t)
2006-10-26
2006
English.
In: BMC Bioinformatics. - : Springer Science and Business Media LLC. - 1471-2105. ; 7, s. 475-
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • Background: Two-Dimensional Difference In Gel Electrophoresis (2D-DIGE) is a powerful tool for measuring differences in protein expression between samples or conditions. However, to remove systematic variability within and between gels the data has to be normalized. In this study we examined the ability of four existing and four novel normalization methods to remove systematic bias in data produced with 2D-DIGE. We also propose a modification of an existing method where the statistical framework determines whether a set of proteins shows an association with the predefined phenotypes of interest. This method was applied to our data generated from a monkey model (Macaca fascicularis) of Parkinson's disease. Results: Using 2D-DIGE we analysed the protein content of the striatum from 6 control and 21 MPTP-treated monkeys, with or without de novo or long-term L-DOPA administration. There was an intensity and spatial bias in the data of all the gels examined in this study. Only two of the eight normalization methods evaluated ('2D loess+scale' and 'SC-2D+quantile') successfully removed both the intensity and spatial bias. In 'SC-2D+quantile' we extended the commonly used loess normalization method against dye bias in two-channel microarray systems to suit systems with three or more channels. Further, by using the proposed method, Differential Expression in Predefined Proteins Sets (DEPPS), several sets of proteins associated with the priming effects of L-DOPA in the striatum in parkinsonian animals were identified. Three of these sets are proteins involved in energy metabolism and one set involved proteins which are part of the microtubule cytoskeleton. Conclusion: Comparison of the different methods leads to a series of methodological recommendations for the normalization and the analysis of data, depending on the experimental design. Due to the nature of 2D-DIGE data we recommend that the p-values obtained in significance tests should be used as rankings only. Individual proteins may be interesting as such, but by studying sets of proteins the interpretation of the results are probably more accurate and biologically informative.

Subject headings

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Farmaceutiska vetenskaper (hsv//swe)
MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Pharmaceutical Sciences (hsv//eng)

Keyword

PHARMACY
FARMACI

Publication and Content Type

ref (subject category)
art (subject category)

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