Molecular and Cellular Biochemistry

, Volume 236, Issue 1–2, pp 139–146 | Cite as

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

  • Elisabeth Deindl
  • Kerstin Boengler
  • Niels van Royen
  • Wolfgang Schaper


Housekeeping genes like glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin are often used as internal standards for quantitative RNA analysis. In our study we analyzed the relative expression level of GAPDH and β-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 β-actin- and GAPDH mRNA during the first 24 h of vessel growth. For day 3 our results revealed an even stronger upregulation of the β-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 β-actin mRNA is an appropriate internal control for relative quantitation of gene expression under conditions of cell proliferation in growing vessels.

GADPH β-actin 18 S rRNA Poly (A)+ RNA relative quantitation of gene expression collateral artery growth 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Spanakis E: Problems related to the interpretation of autoradiographic data on gene expression using common constitutive transcripts as controls. Nucleic Acids Res 21: 3809–3819, 1993PubMedGoogle Scholar
  2. 2.
    Spanakis E, Brouty-Boye D: Evaluation of quantitative variation in gene expression. Nucleic Acids Res 22: 799–806, 1994PubMedGoogle Scholar
  3. 3.
    Bhatia P, Taylor W, Greenberg A, Wright J: Comparison of glycerinaldehyde-3–phosphate dehydrogenase and 28S ribosomal RNA gene expression as RNA loading controls for Northern blot analysis of cell lines of varying malignant potential. Anal Biochem 216: 223–226, 1994PubMedGoogle Scholar
  4. 4.
    Chang T, Juan C, Yin P, Chi C, Tsay H: Up-regulation of beta-actin, cyclophilin and GAPDH in NISI rat hepatoma. Oncol Rep 5: 469–471, 1998PubMedGoogle Scholar
  5. 5.
    de Leeuw W, Siaboom P, Vijg J: Quantitative comparison of mRNA level in mammalian tissue: 28S Ribosomal RNA level as an accurate internal control. Nucleic Acids Res 17: 10137–10138, 1989PubMedGoogle Scholar
  6. 6.
    Foss D, Baarsch M, Murtaugh M: Regulation of hypoxanthine phosphoribosyltransferase, glycerinaldehyde-3–phosphate dehydrogenase and beta-actin mRNA expression in porcine immune cells and tissue. Anim Biotechnol 9: 67–78, 1998PubMedGoogle Scholar
  7. 7.
    Labuhn M, Brack C: Changes in mRNA expression of actin isoforms in Drosophila melanogaster. Gerontology 43: 261–267, 1997PubMedGoogle Scholar
  8. 8.
    Zhong H, Simons JW: Direct comparison of GAPDH, β-actin, cyclophilin, and 28S rRNA as internal standards for quantifying RNA levels under hypoxia. Biophys Res Commun 259: 523–526, 1999Google Scholar
  9. 9.
    Schmittgen TD, Zakrajsek BA: Effect of experimental treatment on housekeeping gene expression: Validation by real-time, quantitative RT-PCR. J Biochem Biophys Meth 46: 69–81, 2000PubMedGoogle Scholar
  10. 10.
    Sharma HS, Wünsch M, Brand T, Verdouw PD, Schaper W: Molecular biology of the coronary vascular and myocardial responses to ischemia. J Cardiovasc Pharmacol 20: S23–S31, 1992Google Scholar
  11. 11.
    Seki N, Kodama J, Hongo A, Miyagi Y, Yoshinouchi M, Kudo T: Vascular endothelial growth factor and platelet-derived endothelial cell growth factor expression are implicated in the angiogenesis of endometrial cancer. Eur J Cancer 36: 68–73, 2000PubMedGoogle Scholar
  12. 12.
    Buschmann I, Schaper W: Arteriogenesis vs. angiogenesis: Two mechanisms of vessel growth. News Physiol Sci 14: 121–125, 1999PubMedGoogle Scholar
  13. 13.
    Deindl E, Schaper W: Collateral and capillary formation - a comparison. In: J.A. Dormandy, W.P. Dole, G.M. Rubanyi (eds). Therapeutic Angiogenesis. Ernst Schering Research Foundation - Workshop 28. Springer, Berlin Heidelberg New York, 1999, pp 67–86Google Scholar
  14. 14.
    van Royen N, Piek JJ, Buschmann I, Hoefer I, Voskuil M, Schaper W: Stimulation of arteriogenesis; a new concept for the treatment of arterial occlusive diseases. Cardiovasc Res 49: 543–553, 2001PubMedGoogle Scholar
  15. 15.
    Plate KH, Breier G, Weich HA, Risau W: Vascular endothelial growth factor is a potential tumor angiogenesis factor in human gliomas in vivo. Nature 359: 845–848, 1992PubMedGoogle Scholar
  16. 16.
    Schaper W, Schaper J: In: Collateral Circulation - Heart, Brain, Kidney, Limbs. Kluwer Academic Publishers, Boston, Dordrecht, London, 1993Google Scholar
  17. 17.
    Folkman J: Angiogenesis in cancer, vascular, rheumatoid and other diseases. Nat Med 1: 27–31, 1995PubMedGoogle Scholar
  18. 18.
    Deindl E, Buschmann I, Hoefer IE, Podzuweit T, Boengler K, Vogel S, van Royen N, Fernandez B, Schaper W: Role of ischemia and hypoxia-inducible genes in arteriogenesis after femoral artery occlusion in the rabbit. Circ Res 89: 779–786, 2001PubMedGoogle Scholar
  19. 19.
    Ito WD, Arras M, Scholz D, Winkler B, Htun P, Schaper W: Angiogenesis but not collateral growth is associated with ischemia after femoral artery occlusion. Am J Physiol 273: H1255–H1265, 1997PubMedGoogle Scholar
  20. 20.
    Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987CrossRefPubMedGoogle Scholar
  21. 21.
    Ross R: The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol 50: 172–186, 1971PubMedGoogle Scholar
  22. 22.
    Sambrook J, Fritsch EF, Maniatis T: In: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989Google Scholar
  23. 23.
    Oberbaeumer I: Retroposons do jump: A B2 element recently integrated in an 18S rDNA gene. Nucleic Acids Res 20: 671–677, 1992PubMedGoogle Scholar
  24. 24.
    Scholz D, Ito W, Fleming I, Deindl E, Sauer A, Wiesnet M, Busse R, Schaper J, Schaper W: Ultrastructure and molecular histology of rabbit hindlimb collateral artery growth (arteriogenesis). Virchows Arch 436: 257–270, 2000PubMedGoogle Scholar
  25. 25.
    Ito WD, Arras M, Winkler B, Scholz D, Schaper J, Schaper W: Monocyte chemotactic protein-1 increases collateral and peripheral conductance after femoral artery occlusion. Circ Res 80: 829–837, 1997PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Elisabeth Deindl
    • 1
  • Kerstin Boengler
    • 1
  • Niels van Royen
    • 1
  • Wolfgang Schaper
    • 1
  1. 1.Department of Experimental CardiologyMax-Planck-InstituteBad NauheimGermany

Personalised recommendations