Regulation of human renin secretion and renin transcription by quantitative PCR in cultured chorionic cells: synergistic effect of cyclic AMP and protein kinase C.
A quantitative PCR was developed to measure human renin mRNA in chorionic cells under various stimuli. An internal standard consisting of a mutated renin mRNA with an insertion of 60 bp was designed to quantify the reaction. Quantitative PCR is a suitable tool for studying human renin gene transcription from a low number of cells. Forskolin alone had no effect on either renin mRNA levels or renin production whereas 10(-7)M PMA stimulated renin mRNA levels and renin production 5- and 2.6-fold, respectively, after 24 hours incubation. PMA and forskolin acted synergistically to increase both renin mRNA levels and renin production in a dose-dependent manner.
INSERM U36-Collège de France, Paris.
N
N
Caroff
R
R
Della Bruna
J
J
Philippe
P
P
Corvol
F
F
Pinet
D001483:Base Sequence; D002478:Cells, Cultured; D002823:Chorion / Q000187:drug effects / Q000201:enzymology*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D011493:Protein Kinase C / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012083:Renin / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D005576:Colforsin; D000242:Cyclic AMP; D011493:Protein Kinase C; D012083:Renin; D013755:Tetradecanoylphorbol Acetate