Abstract
The plasmalemmal Ca(2+) adenosine triphosphatase (PMCA) is a key regulator of Ca(2+) efflux in vascular smooth muscle. In these studies we developed a real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) assay for assessing PMCA1 mRNA levels in rat primary cultured aortic myocytes. This assay detected fetal bovine serum-induced increases in PMCA1 mRNA (relative to 18S rRNA) 4, 8, and 24 h after stimulation. Early fetal bovine serum-induced increases in PMCA1 mRNA were insensitive to the Ca(2+) channel blockers nifedipine, flunarizine, and SKF-96365. These studies demonstrate the feasibility of real-time RT-PCR to assess mRNA levels of PMCA1 and illustrate dynamic regulation of this Ca(2+) pump isoform in rat primary cultured aortic myocytes.
Copyright 2000 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Aorta
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Calcium / metabolism
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Calcium Channel Blockers / pharmacology
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Calcium-Transporting ATPases / antagonists & inhibitors
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Calcium-Transporting ATPases / genetics*
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Calcium-Transporting ATPases / metabolism
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Cation Transport Proteins
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Cattle
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Cells, Cultured
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Flunarizine / pharmacology
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Gene Expression Regulation, Enzymologic / drug effects
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Imidazoles / pharmacology
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Male
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Muscle, Smooth / cytology
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Muscle, Smooth / drug effects
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Muscle, Smooth / enzymology
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Muscle, Smooth / metabolism*
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Nifedipine / pharmacology
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Plasma Membrane Calcium-Transporting ATPases
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Protein Isoforms / antagonists & inhibitors
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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Rats
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Rats, Sprague-Dawley
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Reverse Transcriptase Polymerase Chain Reaction*
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Time Factors
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Up-Regulation / drug effects
Substances
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Calcium Channel Blockers
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Cation Transport Proteins
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Imidazoles
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Protein Isoforms
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RNA, Messenger
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Plasma Membrane Calcium-Transporting ATPases
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Atp2b1 protein, rat
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Calcium-Transporting ATPases
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1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole
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Nifedipine
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Flunarizine
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Calcium