Expression and function of recombinant endothelial nitric oxide synthase gene in canine basilar artery

Alex F.Y. Chen, Timothy O'Brien, Masato Tsutsui, Hiroyuki Kinoshita, Vincent J. Pompili, Thomas B. Crotty, David J. Spector, Zvonimir S. Katusic

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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 colt β-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, N(G)-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.

Original languageEnglish (US)
Pages (from-to)327-335
Number of pages9
JournalCirculation Research
Volume80
Issue number3
DOIs
Publication statusPublished - Jan 1 1997

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Keywords

  • adenovirus vector
  • cerebral artery
  • gene therapy
  • nitric oxide synthase

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Chen, A. F. Y., O'Brien, T., Tsutsui, M., Kinoshita, H., Pompili, V. J., Crotty, T. B., ... Katusic, Z. S. (1997). Expression and function of recombinant endothelial nitric oxide synthase gene in canine basilar artery. Circulation Research, 80(3), 327-335. https://doi.org/10.1161/01.RES.80.3.327