Development of a toxicological gene array and quantitative assessment of this technology

Matthew Bartosiewicz, Mary Trounstine, David Barker, Richard Johnston, Alan R Buckpitt

Research output: Contribution to journalArticlepeer-review

115 Scopus citations

Abstract

High-density arrays of DNA bound to solid substrates offer a powerful approach to identifying changes in gene expression in response to toxicants. While DNA arrays have been used to explore qualitative changes in gene regulation, less attention has focused on the quantitative aspects of this technology. Arrays containing expressed sequence tags for xenobiotic metabolizing enzymes, proteins associated with glutathione regulation, DNA repair enzymes, heat shock proteins, and housekeeping genes were used to examine gene expression in response to β-naphthoflavone (β-NF). Upregulation of cytochrome P4501a1 (Cyp1a1) and 1a2 in mouse liver was maximal 8 h after β-NF administration. Significant upregulation of Cyp1a2 was noted at β-NF doses as low as 0.62 and 1.2 mg/kg when gene expression was measured by microarray or Northern blotting, respectively. Maximal Cyp1a2 induction is 5-fold by Northern analysis and 10-fold by microarray. Induction of Cyp1a1 was 15-and 20-fold by Northern and microarray analysis, respectively. The coefficient of variation for spot to spot and slide to slide comparisons was <15%; this variability was smaller than interanimal variability (18-60%). Comparison of mRNA expression in control animals indicated that there are differences in labeling/detection associated with Cy3/Cy5 dyes; accordingly, experiments must include methods for establishing baseline signals for all genes. We conclude that the dynamic range and sensitivity of DNA microarrays on glass slides is comparable to Northern blotting analysis and that variability of the data introduced during spotting and hybridization is less than the interanimal variability. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)66-73
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume376
Issue number1
DOIs
StatePublished - Apr 1 2000

Keywords

  • β-napthoflavone
  • DNA arrays
  • Gene expression

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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