title: Endothelial cell-derived nitric oxide mobilization is attenuated in copper-deficient rats.
abstract: The attenuation of endothelium-dependent nitric oxide (NO) mediated vasodilation is a consistent finding in both conduit and resistance vessels during dietary copper (Cu) deficiency. Although the effect is well established, evidence for the mechanism remains circumstantial. This study was designed to determine the relative amount of NO produced in and released from the vascular endothelium. Using the fluorescent NO indicator, 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM), we now demonstrate the effect of a Cu-deficient diet on the production of NO from the endothelium of resistance arterioles. In one group of experiments, control and Cu-chelated lung microvascular endothelial cells (ECs) were used to assay NO production and fluorescence was observed by confocal microscopy. Weanling Sprague-Dawley rats were fed purified diets that were either Cu adequate (6.3 micrograms Cu per gram of food) or Cu deficient (0.3 micrograms Cu per gram of food) for 4 weeks. In the second series of experiments, first-order arterioles were microsurgically isolated from the rat cremaster muscle, cannulated, and pressurized with (3[N-morpholino]propanesulfonic acid) physiologic salt solution (MOPS-PSS). DAF-FM (5 micromol.L-1) was added in the lumen of the vessel to measure NO release. Baseline DAF-FM fluorescence was significantly lower in Cu-chelated ECs than in controls. In response to 10-6 mol.L-1 acetylcholine, fluorescent intensity was significantly less in chelated ECs and in the lumen of Cu-deficient arterioles. The results suggest that production and release of NO by the vascular endothelium is inhibited by a restriction of Cu. This inhibition may account for the attenuated vasodilation previously reported in Cu-deficient rats.
authors.affiliation: Department of Physiology and Biophysics, University of Louisville School of Medicine, Louisville, KY 40292, USA.
authors.forename: Jeff C
authors.initials: JC
authors.lastname: Falcone
authors.forename: David
authors.initials: D
authors.lastname: Lominadze
authors.forename: W Thomas
authors.initials: WT
authors.lastname: Johnson
authors.forename: Dale A
authors.initials: DA
authors.lastname: Schuschke
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001160:Arterioles / Q000187:drug effects / Q000378:metabolism; D003300:Copper / Q000008:administration & dosage / Q000172:deficiency*; D004032:Diet / Q000379:methods; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D008297:Male; D008833:Microcirculation / Q000187:drug effects; D018482:Muscle, Skeletal / Q000098:blood supply; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D014664:Vasodilation / Q000187:drug effects
chemical_list: D009569:Nitric Oxide; D003300:Copper