title: Protein expression, vascular reactivity and soluble guanylate cyclase activity in mice lacking the endothelial cell nitric oxide synthase: contributions of NOS isoforms to blood pressure and heart rate control.
abstract: OBJECTIVE
  Both disruption of the endothelial nitric oxide synthase (eNOS) gene and pharmacological inhibition of the NOS produce modest hypertension. It is unclear if and to what extent NOS isoforms other than eNOS contribute to this effect and how loss of one copy of the eNOS gene might impact on vascular reactivity or eNOS protein expression.
  
  
  METHODS
  We examined protein expression, vascular reactivity, activity of soluble guanylate cyclase, blood pressure and heart rate in mice completely lacking the eNOS gene (eNOS-/-), wild-type mice (eNOS+/+) and mice heterozygotic for the eNOS gene (eNOS+/-).
  
  
  RESULTS
  While eNOS-/- mice had mild hypertension and bradycardia, eNOS+/- mice were normotensive. In control mice, oral administration of L-NAME (approximately 100 mg/kg/day x 21 days) increased blood pressure to levels observed in eNOS-/- mice. In eNOS-/- mice, chronic oral administration of L-NAME had no effect on blood pressure, suggesting that inhibition of other NOS isoforms unlikely contribute to hypertension. L-NAME treatment induced bradycardia in both control and eNOS-/- mice, suggesting that both eNOS and other isoforms of NOS might be involved in heart rate control. Studies of aortic rings from eNOS-/- mice revealed a complete lack of endothelium-dependent vascular relaxation in response to acetylcholine and the calcium ionophore A23187 and an increase in sensitivity to phenylephrine, serotonin and nitroglycerin. Aortic rings from eNOS+/- mice demonstrated only minor alterations of responses to nitroglycerin and a normal relaxation to either acetylcholine or A23187 compared to vessels from eNOS-/+. Western analysis demonstrated that eNOS expression was virtually identical between eNOS+/+ and eNOS+/- mice and was absent in eNOS-/- mice. The activity of lung-isolated soluble guanylate cyclase was identical in the three strains of mice.
  
  
  CONCLUSIONS
  We conclude that loss of one copy of the eNOS gene, as observed in heterozygotic animals, has no effect on vascular reactivity, blood pressure or eNOS protein expression. Isoforms of NOS, other than eNOS are unlikely involved in blood pressure regulation but may participate in heart rate control.
authors.affiliation: Department of Medicine, Emory University School of Medicine, Atlanta, GA, USA. kojda@uni-duesseldorf.de
authors.forename: G
authors.initials: G
authors.lastname: Kojda
authors.forename: J B
authors.initials: JB
authors.lastname: Laursen
authors.forename: S
authors.initials: S
authors.lastname: Ramasamy
authors.forename: J D
authors.initials: JD
authors.lastname: Kent
authors.forename: S
authors.initials: S
authors.lastname: Kurz
authors.forename: J
authors.initials: J
authors.lastname: Burchfield
authors.forename: E G
authors.initials: EG
authors.lastname: Shesely
authors.forename: D G
authors.initials: DG
authors.lastname: Harrison
mesh_terms: D000818:Animals; D001013:Aorta, Thoracic; D001794:Blood Pressure / Q000235:genetics; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017353:Gene Deletion; D015870:Gene Expression; D006162:Guanylate Cyclase / Q000378:metabolism; D006339:Heart Rate / Q000235:genetics; D006439:Hemodynamics / Q000235:genetics*; D006579:Heterozygote; D006973:Hypertension / Q000235:genetics* / Q000378:metabolism; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D014661:Vasoconstriction / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D006162:Guanylate Cyclase; D019331:NG-Nitroarginine Methyl Ester