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Am J Physiol Heart Circ Physiol 273: H2575-H2579, 1997;
0363-6135/97 $5.00
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Vol. 273, Issue 6, H2575-H2579, December 1997

Induction of nitric oxide synthase mRNA in coronary resistance arteries isolated from exercise-trained pigs

Christopher R. Woodman, Judy M. Muller, M. Harold Laughlin, and Elmer M. Price

Department of Veterinary Biomedical Sciences and Dalton Cardiovascular Research Center, University of Missouri, Columbia, Missouri 65211

The purpose of this study was to develop a method by which endothelial cell nitric oxide synthase (ecNOS) mRNA expression could be measured in single coronary resistance arteries and to test the hypothesis that ecNOS gene expression is upregulated by exercise training. Yucatan miniature swine were randomly assigned to exercise-trained (ET; n = 5) or sedentary (Sed; n = 4) groups for 16 wk. Individual coronary resistance arteries (50-100 µm) were dissected, frozen in liquid nitrogen, and homogenized in a LiCl buffer. mRNA was isolated from each vessel, and ecNOS gene expression was assessed using reverse transcriptase (RT)-polymerase chain reaction (PCR) standardized by coamplifying ecNOS with glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The ecNOS-to-GAPDH amplicon ratio was significantly greater in coronary resistance arteries isolated from ET pigs than in Sed controls. On the basis of these data, it is concluded that RT-PCR can be used on single coronary resistance arteries to assess cell-specific mRNA expression and that ecNOS gene expression is upregulated by exercise training in porcine coronary resistance arteries.

gene expression; reverse-transcription polymerase chain reaction; cDNA synthesis; endothelium-derived relaxing factor


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