This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Akiyama, M. Articles by Katusic, Z. S. Search for Related Content PubMed PubMed Citation Articles by Akiyama, M. Articles by Katusic, Z. S. Related Collections Endothelium/vascular type/nitric oxide

(Stroke. 2002;33:1071.)
© 2002 American Heart Association, Inc.

Original Contributions

Expression and Function of Recombinant S1179D Endothelial Nitric Oxide Synthase in Canine Cerebral Arteries

Masahiko Akiyama, MD; Daihiko Eguchi, MD, PhD; Deborah Weiler, BA, MS; Timothy O’Brien, MD; Imre Kovesdi, PhD; Ramona S. Scotland, PhD; William C. Sessa, PhD Zvonimir S. Katusic, MD, PhD

From the Departments of Anesthesiology and Molecular Pharmacology and Experimental Therapeutics (M.A., D.E., D.W., Z.S.K.) and Division of Endocrinology and Metabolism (T.O’B.), Mayo Clinic and Foundation, Rochester, Minn; GenVec, Inc, Gaithersburg, Md (I.K.); and Department of Pharmacology and Molecular Cardiobiology Program, Yale University School of Medicine, New Haven, Conn (R.S.S., W.C.S.).

Correspondence to Zvonimir S. Katusic, MD, PhD, Department of Anesthesiology, Mayo Clinic, 200 First St SW, Rochester, MN 55905. E-mail katusic.zvonimir{at}mayo.edu

Background and Purpose— Bovine endothelial nitric oxide synthase (eNOS) is phosphorylated directly by the protein kinase Akt at serine 1179. Mutation of this residue to the negatively charged aspartate (S1179DeNOS) increases nitric oxide (NO) production constitutively in the absence of agonist stimulus. The present study was designed to determine the effect of mutant S1179DeNOS gene expression on vasomotor function of canine cerebral arteries.

Methods— Isolated basilar and middle cerebral arteries were exposed ex vivo (30 minutes at 37°C) to an adenoviral vector (10¹⁰ plaque-forming units per milliliter) encoding the S1179DeNOS gene (AdCMVS1179DeNOS), the wild-type eNOS gene (AdCMVeNOS), or the green fluorescent protein (GFP) reporter gene (AdCMVGFP). Twenty-four hours after transduction, arteries were suspended in an organ chamber for isometric force recording, and levels of cGMP were measured by radioimmunoassay.

Results— Transgene protein expression was detected mainly in the vascular adventitia. In AdCMVS1179DeNOS-transduced arteries, basal levels of cGMP were significantly elevated compared with those in control (nontransduced), AdCMVGFP-, or AdCMVeNOS-transduced vessels (n=8; P<0.01). The elevation of cGMP was abolished by a NOS inhibitor, N^G-nitro-L-arginine methyl ester (L-NAME), or by incubation in the calcium-free medium in the presence of calcium chelators. In AdCMVS1179DeNOS-transduced arteries, contractions to endothelin-1 (10^-10 to 10^-8 mol/L) were significantly reduced compared with those in control and AdCMVGFP-transduced arteries (n=7; P<0.05). The vasoconstrictor effect of endothelin-1 was restored in the presence of the NOS inhibitor L-NAME.

Conclusions— Our results suggest that in cerebral arteries, expression of recombinant S1179DeNOS increases basal production of NO and inhibits the vasoconstrictor effect of endothelin-1. This effect may have therapeutic application in prevention and treatment of cerebrovascular diseases.

Key Words: cerebral arteries • gene transfer • nitric oxide • dogs

This article has been cited by other articles:

				A. V. R. Santhanam, L. A. Smith, T. He, K. A. Nath, and Z. S. Katusic Endothelial Progenitor Cells Stimulate Cerebrovascular Production of Prostacyclin By Paracrine Activation of Cyclooxygenase-2 Circ. Res., May 11, 2007; 100(9): 1379 - 1388. [Abstract] [Full Text] [PDF]

				A. V. R. Santhanam, L. A. Smith, K. A. Nath, and Z. S. Katusic In vivo stimulatory effect of erythropoietin on endothelial nitric oxide synthase in cerebral arteries Am J Physiol Heart Circ Physiol, August 1, 2006; 291(2): H781 - H786. [Abstract] [Full Text] [PDF]

				A. V. R. Santhanam, L. A. Smith, M. Akiyama, A. G. Rosales, K. R. Bailey, and Z. S. Katusic Role of Endothelial NO Synthase Phosphorylation in Cerebrovascular Protective Effect of Recombinant Erythropoietin During Subarachnoid Hemorrhage- Induced Cerebral Vasospasm Stroke, December 1, 2005; 36(12): 2731 - 2737. [Abstract] [Full Text] [PDF]

				J. Yu, E. D. deMuinck, Z. Zhuang, M. Drinane, K. Kauser, G. M. Rubanyi, H. S. Qian, T. Murata, B. Escalante, and W. C. Sessa Endothelial nitric oxide synthase is critical for ischemic remodeling, mural cell recruitment, and blood flow reserve PNAS, August 2, 2005; 102(31): 10999 - 11004. [Abstract] [Full Text] [PDF]

				O. Suda, L. A. Smith, L. V. d'Uscio, T. E. Peterson, and Z. S. Katusic In Vivo Expression of Recombinant Vascular Endothelial Growth Factor in Rabbit Carotid Artery Increases Production of Superoxide Anion Arterioscler. Thromb. Vasc. Biol., March 1, 2005; 25(3): 506 - 511. [Abstract] [Full Text] [PDF]

				J. Sorenson, A. V. R. Santhanam, L. A. Smith, M. Akiyama, W. C. Sessa, and Z. S. Katusic Expression and Function of Recombinant S1179D Endothelial NO Synthase in Human Pial Arteries Stroke, January 1, 2005; 36(1): 158 - 160. [Abstract] [Full Text] [PDF]

				Z. S. Katusic, N. M. Caplice, and K. A. Nath Nitric Oxide Synthase Gene Transfer as a Tool to Study Biology of Endothelial Cells Arterioscler. Thromb. Vasc. Biol., November 1, 2003; 23(11): 1990 - 1994. [Abstract] [Full Text] [PDF]

				D. Eguchi, L. V. d'Uscio, C. Wambi, D. Weiler, I. Kovesdi, T. O'Brien, and Z. S. Katusic Inhibitory effect of recombinant iNOS gene expression on vasomotor function of canine basilar artery Am J Physiol Heart Circ Physiol, December 1, 2002; 283(6): H2560 - H2566. [Abstract] [Full Text] [PDF]