Differential expression of GAPDH and beta3-actin in growing collateral arteries.
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.
Deindl
Elisabeth
E
Max-Planck-Institute, Department of Experimental Cardiology, Bad Nauheim, Germany. e.deindl@kerckhoff.mpg.de
Boengler
Kerstin
K
van Royen
Niels
N
Schaper
Wolfgang
W
D000199:Actins / Q000096:biosynthesis* / Q000378:metabolism; D000818:Animals; D001158:Arteries / Q000201:enzymology* / Q000378:metabolism*; D015152:Blotting, Northern; D002455:Cell Division; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D005786:Gene Expression Regulation; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis*; D019394:Ki-67 Antigen / Q000096:biosynthesis; D008856:Microscopy, Fluorescence; D011061:Poly A / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012337:RNA, Ribosomal, 18S / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014018:Tissue Distribution
D000199:Actins; D018076:DNA, Complementary; D019394:Ki-67 Antigen; D012337:RNA, Ribosomal, 18S; D011061:Poly A; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases