Microarray analysis of differential gene expression in lead-exposed astrocytes

Toxicol Appl Pharmacol. 2001 Oct 1;176(1):34-53. doi: 10.1006/taap.2001.9274.

Abstract

The toxic metal lead is a widespread environmental health hazard that can adversely affect human health. In an effort to better understand the cellular and molecular consequences of lead exposure, we have employed cDNA microarrays to analyze the effects of acute lead exposure on large-scale gene expression patterns in immortalized rat astrocytes. Our studies identified many genes previously reported to be differentially regulated by lead exposure. Additionally, we have identified novel putative targets of lead-mediated toxicity, including members of the family of calcium/phospholipid binding annexins, the angiogenesis-inducing thrombospondins, collagens, and tRNA synthetases. We demonstrate the ability to distinguish lead-exposed samples from control or sodium samples solely on the basis of large-scale gene expression patterns using two complementary clustering methods. We have confirmed the altered expression of candidate genes and their encoded proteins by RT-PCR and Western blotting, respectively. Finally, we show that the calcium-dependent phospholipid binding protein annexin A5, initially identified as a differentially regulated gene by our microarray analysis, is directly bound and activated by nanomolar concentrations of lead. We conclude that microarray technology is an effective tool for the identification of lead-induced patterns of gene expression and molecular targets of lead.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acyl-tRNA Synthetases / genetics
  • Animals
  • Annexins / genetics
  • Annexins / metabolism
  • Astrocytes / drug effects*
  • Astrocytes / metabolism*
  • Blotting, Western
  • Cell Line, Transformed
  • Collagen / genetics
  • DNA, Complementary / analysis
  • Gene Expression Regulation / drug effects*
  • Lead / metabolism
  • Lead / pharmacology*
  • Lead / toxicity
  • Oligonucleotide Array Sequence Analysis*
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thrombospondins / genetics

Substances

  • Annexins
  • DNA, Complementary
  • Thrombospondins
  • Lead
  • Collagen
  • Amino Acyl-tRNA Synthetases