Differential expression of GAPDH and beta3-actin in growing collateral arteries

Mol Cell Biochem. 2002 Jul;236(1-2):139-46. doi: 10.1023/a:1016166127465.

Abstract

Housekeeping genes like glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin are often used as internal standards for quantitative RNA analysis. In our study we analyzed the relative expression level of GAPDH and beta-actin as well as of the 18S rRNA and the Poly (A)+ RNA in growing collateral arteries in a rabbit model of arteriogenesis which is not associated with ischemia. Relative quantitation of the housekeeping genes displayed a significant upregulation of the beta-actin- and GAPDH mRNA during the first 24 h of vessel growth. For day 3 our results revealed an even stronger upregulation of the beta-actin mRNA (140%) but a significant downregulation of the GAPDH mRNA (50% of control). The 18S rRNA, however, showed for the same periods only minor alterations compared to the Poly (A)+ RNA. From these results we conclude that the 18S rRNA, but not the GAPDH- or beta-actin mRNA is an appropriate internal control for relative quantitation of gene expression under conditions of cell proliferation in growing vessels.

MeSH terms

  • Actins / biosynthesis*
  • Actins / metabolism
  • Animals
  • Arteries / enzymology*
  • Arteries / metabolism*
  • Blotting, Northern
  • Cell Division
  • Cells, Cultured
  • DNA, Complementary / metabolism
  • Gene Expression Regulation
  • Glyceraldehyde-3-Phosphate Dehydrogenases / biosynthesis*
  • Ki-67 Antigen / biosynthesis
  • Microscopy, Fluorescence
  • Poly A / metabolism
  • RNA / metabolism
  • RNA, Ribosomal, 18S / metabolism
  • Rabbits
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Tissue Distribution

Substances

  • Actins
  • DNA, Complementary
  • Ki-67 Antigen
  • RNA, Ribosomal, 18S
  • Poly A
  • RNA
  • Glyceraldehyde-3-Phosphate Dehydrogenases