Endothelial nitric oxide synthase gene transfer enhances dilation of newborn piglet pulmonary arteries

Judy L. Aschner, Nora Kovacs, James V. Perciaccante, Jorge P. Figueroa, Nishadi Thrikawala, Gregory S. Robins, David W. Busija

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We determined the expression and functional correlate of in vitro transfection with a recombinant adenoviral vector encoding the gene for bovine endothelial nitric oxide synthase (AdCMVeNOS) or Escherichia coli β- galactosidase (AdCMVLacZ) in pulmonary endothelial cells (EC), vascular smooth muscle cells (VSMC), and pulmonary arteries (PA) from newborn piglets. AdCMVeNOS and AdCMVeLacZ vectors, grown in 293-cell monolayers, were purified by double-cesium gradient ultracentrifugation. Cell cultures and PA were incubated with increasing vector titers for 30 or 60 min, followed by incubation in fresh medium for 18 h at 37°C. LacZ expression was assessed by histochemical staining; eNOS expression was evaluated by Western blot analysis. Functional eNOS expression was determined by measurement of cGMP and quantification of the relaxation response to bradykinin (BK). In PA, LacZ transgene expression was preferentially localized to the adventitia and endothelium. Increased eNOS protein expression was observed in EC and VSMC transfected with AdCMVeNOS. Functional studies revealed increased cGMP abundance in cultured cells and enhanced relaxation to BK in AdCMVeNOS- transfected PA. These studies demonstrate that gene transfer with AdCMVeNOS results in functional expression and altered vasoactive responses in the neonatal pulmonary vasculature. Gene transfer with replication-deficient adenovirus vectors is a useful tool for the study of targeted genes in vascular biology.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume277
Issue number1 46-1
StatePublished - Jul 1999
Externally publishedYes

Fingerprint

Nitric Oxide Synthase Type III
Pulmonary Artery
Dilatation
Bradykinin
Vascular Smooth Muscle
Genes
Smooth Muscle Myocytes
Endothelial Cells
Galactosidases
Adventitia
Lung
Cesium
Ultracentrifugation
Transgenes
Adenoviridae
Endothelium
Transfection
Blood Vessels
Cultured Cells
Cell Culture Techniques

Keywords

  • Adenovirus
  • Endothelium
  • Nitric oxide
  • Persistent pulmonary hypertension of the newborn

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Aschner, J. L., Kovacs, N., Perciaccante, J. V., Figueroa, J. P., Thrikawala, N., Robins, G. S., & Busija, D. W. (1999). Endothelial nitric oxide synthase gene transfer enhances dilation of newborn piglet pulmonary arteries. American Journal of Physiology - Heart and Circulatory Physiology, 277(1 46-1).

Endothelial nitric oxide synthase gene transfer enhances dilation of newborn piglet pulmonary arteries. / Aschner, Judy L.; Kovacs, Nora; Perciaccante, James V.; Figueroa, Jorge P.; Thrikawala, Nishadi; Robins, Gregory S.; Busija, David W.

In: American Journal of Physiology - Heart and Circulatory Physiology, Vol. 277, No. 1 46-1, 07.1999.

Research output: Contribution to journalArticle

Aschner, Judy L. ; Kovacs, Nora ; Perciaccante, James V. ; Figueroa, Jorge P. ; Thrikawala, Nishadi ; Robins, Gregory S. ; Busija, David W. / Endothelial nitric oxide synthase gene transfer enhances dilation of newborn piglet pulmonary arteries. In: American Journal of Physiology - Heart and Circulatory Physiology. 1999 ; Vol. 277, No. 1 46-1.
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