Transcription of actin, cyclophilin and glyceraldehyde phosphate dehydrogenase genes: tissue- and treatment-specificity.
Studies involving RNA transcription, in varying biological systems, usually necessitate a term of transcriptional reference. Traditionally, the transcription of the gene of interest was compared to a constitutively expressed 'control' gene. Run-on transcription analysis was undertaken to evaluate and compare the transcription of three frequently used 'control genes' (beta-actin, cyclophilin and glyceraldehyde-3-phosphate dehydrogenase), in nine rat tissues. Similarities, but also clear and highly significant differences, were found in the transcription profiles of these three genes. There was significantly greater transcription for uterine glyceraldehyde-phosphate dehydrogenase compared to all other tissues tested, while both cyclophilin and glyceraldehyde-phosphate dehydrogenase were significantly elevated in the adrenal cortex. Upon cholinergic agonist treatment, both beta-actin and glyceraldehyde-phosphate dehydrogenase RNA expression were greatly induced in the adrenal medulla (41- and 94-fold, respectively), while cyclophilin transcription was not altered. In another treatment paradigm, surgical ovariectomy, only uterine glyceraldehyde-phosphate dehydrogenase transcription was significantly reduced. While, all three of these genes are assumed to be constitutively expressed throughout the body and hence used as normalization controls, the current study questions these accepted terms of reference. As cyclophilin transcription was not affected in both treatment paradigms, it should be considered more seriously as a RNA normalization control.
Weisinger
G
G
Department of Endocrinology, Sourasky Medical Center, Tel Aviv, Israel. gweisinger@hotmail.com
Gavish
M
M
Mazurika
C
C
Zinder
O
O
D000199:Actins / Q000235:genetics*; D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism; D000313:Adrenal Medulla / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D018679:Cholinergic Agonists / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D010052:Ovariectomy; D019696:Peptidylprolyl Isomerase / Q000235:genetics*; D012313:RNA / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic; D014599:Uterus / Q000187:drug effects / Q000378:metabolism
D000199:Actins; D018679:Cholinergic Agonists; D004958:Estradiol; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D019696:Peptidylprolyl Isomerase