Real-time polymerase chain reaction to quantify mRNA for endothelial nitric oxide synthase.
The endothelial isoform of nitric oxide synthase (eNOS) plays a major role in vascular biology and emerging evidence suggests it may serve diverse biological functions in nonvascular cells. To be able to study the eNOS at the molecular level, including regulation of mRNA levels, we developed real-time reverse transcription-polymerase chain reaction methodology for sensitive and precise quantification of mRNA levels. In addition to providing quantification of mRNA levels, the method should be attractive because of its high sensitivity and the fact that blood vessels from small species such as genetically altered mice can be studied. Here, we demonstrate that the method can measure mRNA levels in cerebral and coronary arteries from mice, in addition to larger blood vessels.
Chu
Yi
Y
Cardiovascular Center, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, USA.
Faraci
Frank M
FM
D000818:Animals; D001013:Aorta, Thoracic / Q000201:enzymology*; D002339:Carotid Arteries / Q000201:enzymology*; D017931:DNA Primers; D051379:Mice; D008810:Mice, Inbred C57BL; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000302:isolation & purification*
D017931:DNA Primers; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse