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J Appl Physiol (March 7, 2003). doi:10.1152/japplphysiol.00061.2003
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Submitted on January 22, 2003
Accepted on March 3, 2003

Gender, Exercise Training, and eNOS expression in Porcine Skeletal Muscle Arteries

M. H. Laughlin1*, Wade V. Welshons2, Michael Sturek3, James W. E. Rush4, James R. Turk2, Julia A. Taylor2, Barbara M. Judy2, Kyle K. Henderson2, and V K Ganjam2

1 Biomedical Sciences, University of Missouri, Columbia, MO, USA; Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, USA
2 Biomedical Sciences, University of Missouri, Columbia, MO, USA
3 Departments of Medical Pharmacology and Physiology and Internal Medicine, University of Missouri, Columbia, MO, USA
4 Department of Kinesiology, University of Waterloo, Waterloo, ON, Canada

* To whom correspondence should be addressed. E-mail: laughlinm{at}missouri.edu.

The purpose of the present study was to determine the effects of gender and exercise training on endothelial nitric oxide synthase (eNOS) and superoxide dismutase protein (SOD) protein content of porcine skeletal muscle arteries and to evaluate the role of 17{beta}-estradiol (E2) in these effects. We tested the hypothesis that endothelial cells of female arteries express more eNOS and/or SOD than male endothelial cells by measuring protein content with immunoblots and immunohistochemistry in femoral and brachial arteries of male (M) and female (F) pigs. Second, we measured eNOS and SOD content of these arteries from trained M and F pigs to test the hypothesis that exercise training increases eNOS and/or SOD content of these arteries. We also measured estrogen receptor (ER{alpha})mRNA in aortic endothelial cells and ER{alpha} and ER{beta} protein in aortas of M and F pigs. Results indicate that F arteries contain more eNOS than M arteries and that exercise training increases eNOS content independent of gender. M and F pigs expressed similar levels of ER{alpha}mRNA and protein and similar amounts ER{beta} protein in their arteries. E2 concentrations as measured by radioimmunoassay (RIA) were 180 ± 34 pg/ml in M sera, and approximately 5 pg/ml in F sera and neither was changed by training. However, bioassay indicated that biologically active estrogen equivalent to an E2 concentration of only 35 ± 5 pg/ml was present in M sera. E2 in F pigs, whether measured by RIA or bioassay, was approximately 24 pg/ml at peak estrous, and approximately 2 pg/ml on day 5 diestrus. Free serum E2 was lower in M than in F pigs and exercise training increased free E2 in M and decreased it in F. We conclude that: 1) Gender has significant influence on eNOS and SOD content of porcine skeletal muscle arteries., 2) The effects of gender and exercise training vary among arteries of different anatomic origin, 3) That M sera contains compounds that cause RIA to overestimate circulating estrogenic activity., and 4) Relative to the human M, the M pig is not biologically estrogenized by high levels of E2 reported by RIA, while in F pig E2 levels are lower than in the blood of human Fs.




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