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 Onoue, H. Articles by Faraci, F. M. Search for Related Content PubMed PubMed Citation Articles by Onoue, H. Articles by Faraci, F. M. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Medline Plus Health Information Genes and Gene Therapy

(Stroke. 1998;29:1959-1966.)
© 1998 American Heart Association, Inc.

Original Contributions

Expression and Function of Recombinant Endothelial Nitric Oxide Synthase Gene in Canine Basilar Artery After Experimental Subarachnoid Hemorrhage

Hisashi Onoue, MD, PhD; Masato Tsutsui, MD; Leslie Smith; Adele Stelter, BS; Timothy O'Brien, MD; Zvonimir S. Katusic, MD, PhD

From the Departments of Anesthesiology and Pharmacology and Division of Endocrinology and Metabolism (T. O'B.), Mayo Medical Center, Rochester, Minn.

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

Background and Purpose—Gene transfer with recombinant viral vectors encoding vasodilator proteins may be useful in therapy of cerebral vasospasm after subarachnoid hemorrhage (SAH). Relaxations mediated by nitric oxide are impaired in cerebral arteries affected by SAH. The present study was designed to determine the effect of SAH on the efficiency of ex vivo adenovirus-mediated gene transfer to canine basilar arteries and to examine whether expression of recombinant endothelial nitric oxide synthase (eNOS) gene may have functional effects on vasomotor reactivity of spastic arteries affected by SAH.

Methods—Replication-deficient recombinant adenovirus vectors encoding bovine eNOS (AdCMVeNOS) and Escherichia coli ß-galactosidase (AdCMVß-Gal) genes were used for ex vivo gene transfer. Rings of basilar arteries obtained from control dogs and dogs exposed to SAH were incubated with the vectors in minimum essential medium. Twenty-four hours after gene transfer, expression and function of the recombinant genes were evaluated by (1) histochemical or immunohistochemical staining, (2) ß-galactosidase protein measurement, and (3) isometric tension recording.

Results—Transduction with AdCMVß-Gal and AdCMVeNOS resulted in the expression of recombinant ß-galactosidase and eNOS proteins mostly in the vascular adventitia. The expression of ß-galactosidase protein was 2-fold higher in SAH arteries than in normal arteries. Endothelium-dependent relaxations caused by bradykinin and substance P were suppressed in SAH arteries. The relaxations to bradykinin were significantly augmented in both normal and SAH arteries after AdCMVeNOS transduction but not after AdCMVß-Gal transduction. The relaxations to substance P were augmented by AdCMVeNOS transduction only in normal arteries. Bradykinin and substance P caused relaxations even in endothelium-denuded arteries, when the vessels were transduced with AdCMVeNOS. These endothelium-independent (adventitia-dependent) relaxations to bradykinin observed after AdCMVeNOS transduction were similar between normal and SAH arteries, whereas those to substance P were significantly reduced in SAH arteries compared with normal arteries.

Conclusions—These results suggest that expression of recombinant proteins after adenovirus-mediated gene transfer may be enhanced in cerebral arteries affected by SAH and that successful eNOS gene transfer to spastic arteries can at least partly restore the impaired nitric oxide–mediated relaxations through local (adventitial) production of nitric oxide.

Editorial Comment

Frank M. Faraci, , PhD, Guest Editor

Department of Internal Medicine Cardiovascular Division University of Iowa College of Medicine Iowa City, Iowa

This article has been cited by other articles:

				T. Ogawa, S. Ono, T. Ichikawa, S. Arimitsu, K. Onoda, K. Tokunaga, K. Sugiu, K. Tomizawa, H. Matsui, and I. Date Novel Protein Transduction Method by Using 11R: An Effective New Drug Delivery System for the Treatment of Cerebrovascular Diseases Stroke, April 1, 2007; 38(4): 1354 - 1361. [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]

				Y. Watanabe and D. D. Heistad Targeting cerebral arteries for gene therapy Exp Physiol, May 1, 2005; 90(3): 327 - 331. [Abstract] [Full Text] [PDF]

				M. Zanetti, L. V. d'Uscio, I. Kovesdi, Z. S. Katusic, and T. O'Brien In Vivo Gene Transfer of Inducible Nitric Oxide Synthase to Carotid Arteries From Hypercholesterolemic Rabbits Stroke, May 1, 2003; 34(5): 1293 - 1298. [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]

				V. G. Khurana, L. A. Smith, T. A. Baker, D. Eguchi, T. O'Brien, and Z. S. Katusic Protective Vasomotor Effects of In Vivo Recombinant Endothelial Nitric Oxide Synthase Gene Expression in a Canine Model of Cerebral Vasospasm Stroke, March 1, 2002; 33(3): 782 - 789. [Abstract] [Full Text] [PDF]

				S. P. Didion, C. A. Hathaway, and F. M. Faraci Superoxide levels and function of cerebral blood vessels after inhibition of CuZn-SOD Am J Physiol Heart Circ Physiol, October 1, 2001; 281(4): H1697 - H1703. [Abstract] [Full Text] [PDF]

				R. Morishita Adventure of Gene Therapy Into the Brain: A New Era for Cardiovascular Gene Therapy Circ. Res., October 27, 2000; 87(9): 719 - 721. [Full Text] [PDF]

				K. Toyoda, F. M. Faraci, Y. Watanabe, T. Ueda, J. J. Andresen, Y. Chu, S. Otake, and D. D. Heistad Gene Transfer of Calcitonin Gene-Related Peptide Prevents Vasoconstriction After Subarachnoid Hemorrhage Circ. Res., October 27, 2000; 87(9): 818 - 824. [Abstract] [Full Text] [PDF]

				D. D. Lund, F. M. Faraci, F. J. Miller Jr, and D. D. Heistad Gene Transfer of Endothelial Nitric Oxide Synthase Improves Relaxation of Carotid Arteries From Diabetic Rabbits Circulation, March 7, 2000; 101(9): 1027 - 1033. [Abstract] [Full Text] [PDF]

				D. D. Gutterman Adventitia-dependent influences on vascular function Am J Physiol Heart Circ Physiol, October 1, 1999; 277(4): H1265 - H1272. [Full Text] [PDF]

				H. C. Champion, T. J. Bivalacqua, F. M. D'Souza, L. A. Ortiz, J. R. Jeter, K. Toyoda, D. D. Heistad, A. L. Hyman, and P. J. Kadowitz Gene Transfer of Endothelial Nitric Oxide Synthase to the Lung of the Mouse In Vivo : Effect on Agonist-Induced and Flow-Mediated Vascular Responses Circ. Res., June 25, 1999; 84(12): 1422 - 1432. [Abstract] [Full Text] [PDF]

				M. Tsutsui, H. Onoue, Y. Iida, L. Smith, T. O'Brien, and Z. S. Katusic Adventitia-dependent relaxations of canine basilar arteries transduced with recombinant eNOS gene Am J Physiol Heart Circ Physiol, June 1, 1999; 276(6): H1846 - H1852. [Abstract] [Full Text] [PDF]