Expression and Function of Recombinant Endothelial Nitric Oxide Synthase Gene in Canine Basilar Artery
- 1 March 1997
- journal article
- research article
- Published by Ovid Technologies (Wolters Kluwer Health) in Circulation Research
- Vol. 80 (3), 327-335
- https://doi.org/10.1161/01.res.80.3.327
Abstract
Endothelial NO synthase (eNOS) is an enzyme responsible for the production of a potent vasodilator and a key regulator of vascular tone, NO. In peripheral arteries, expression of a recombinant eNOS gene increases production of NO in the blood vessel wall. This approach appears to be a promising strategy for gene therapy of cerebrovascular disease. The major objective of the present study was to determine whether a recombinant eNOS gene (AdCMVNOS) can be functionally expressed in cerebral arteries. Replication-defective recombinant adenovirus vectors encoding bovine eNOS and Escherichia coli beta-galactosidase (AdCMVLacZ) genes, driven by the cytomegalovirus promoter, were used for ex vivo gene transfer. Rings of canine basilar artery were incubated with increasing titers of the vectors in MEM. Twenty-four or forty-eight hours after gene transfer, expression and function of AdCMVNOS were evaluated by (1) immunohistochemical staining, (2) isometric tension recording, and (3) cGMP radioimmunoassay. Transfection with AdCMVNOS resulted in the expression of recombinant eNOS protein in the vascular adventitia and endothelium, associated with significantly reduced contractile responses to UTP and enhanced endothelium-dependent relaxation to calcium ionophore A23187. Basal production of cGMP was significantly increased in the transfected vessels. The reduced contractions to UTP with increased cGMP production were reversed by a NOS inhibitor, NG-monomethyl-L-arginine. Contractions to UTP or production of cGMP were not affected in arteries transfected with AdCMVLacZ reporter gene. The results of the present study represent the first successful transfer and functional expression of recombinant eNOS gene in cerebral arteries. Our findings suggest that cerebral arterial tone can be modulated by recombinant eNOS expression in the vessel wall. (Circ Res. 1997;80:327-335.)Keywords
This publication has 27 references indexed in Scilit:
- Loss of nitric oxide synthase immunoreactivity in cerebral vasospasmJournal of Neurosurgery, 1996
- Effect of intracarotid nitric oxide on primate cerebral vasospasm after subarachnoid hemorrhageJournal of Neurosurgery, 1995
- 1012-102 Alteration of Rabbit Carotid Artery Vasomotor Function by Gene Transfer with a Replication Deficient AdenovirusJournal of the American College of Cardiology, 1995
- Complex Models for the Study of Gene Function in Cardiovascular BiologyAnnual Review of Physiology, 1994
- NITRIC OXIDE: A Physiologic Messenger MoleculeAnnual Review of Biochemistry, 1994
- Combined effect of L-arginine and superoxide dismutase on the spastic basilar artery after subarachnoid hemorrhage in dogsJournal of Neurosurgery, 1994
- Antisense c-myb oligonucleotides inhibit intimal arterial smooth muscle cell accumulation in vivoNature, 1992
- Intravenous nitroglycerin for the treatment of chronic cerebral vasoconstriction in the primateJournal of Neurosurgery, 1981
- Structure-activity relation of pyrimidine nucleotides and nucleoside in canine isolated cerebral vesselsJournal of Pharmacy and Pharmacology, 1978
- Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5Journal of General Virology, 1977