In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs

S. R. Severson, A. Jeppsson, T. O'Brien, C. Pellegrini, Virginia M Miller, H. D. Tazelaar, C. G A McGregor

Research output: Contribution to journalArticle

Abstract

Experiments were designed to study functional markers for endothelial constitutive nitric oxide synthase (ecNOS) after transbronchial adenoviral-mediated (AD) transfer to transplanted lungs. Syngenic orthotopic single lung transplantation in rats was performed after airway administration of either AdecNOS or the marker gene AdLacZ to the donor lung (300 ul, 1x109 pfu/ml; n=4 each). Transplanted lungs were harvested at four days and transgene DNA and mRNA were identified by polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), respectively. Calcium-dependent NOS activity increased in ecNOS compared to LacZ transduced lungs. However, cyclic GMP isolated from lung homogenates was the same in both groups. Positive immunohistochemical staining for ecNOS was observed in pneumocytes only in ecNOS transduced lungs. Preliminary data indicates that oxidized products of nitric oxide (NOx) increased in pulmonary venous blood in ecNOS transduced lungs. These results indicate an effective transduction of pneumocytes with ecNOS that does not alter basal cyclic GMP levels, although NOS protein and nitric oxide does increase.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number5
StatePublished - Mar 20 1998

Fingerprint

Nitric Oxide Synthase Type III
lungs
Lung
Polymerase chain reaction
Cyclic GMP
Alveolar Epithelial Cells
pneumocytes
cyclic GMP
Nitric Oxide
nitric oxide
RNA-Directed DNA Polymerase
Lung Transplantation
endothelial nitric oxide synthase
Rats
Blood
Genes
Reverse Transcriptase Polymerase Chain Reaction
Transgenes
Calcium
Messenger RNA

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Severson, S. R., Jeppsson, A., O'Brien, T., Pellegrini, C., Miller, V. M., Tazelaar, H. D., & McGregor, C. G. A. (1998). In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs. FASEB Journal, 12(5).

In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs. / Severson, S. R.; Jeppsson, A.; O'Brien, T.; Pellegrini, C.; Miller, Virginia M; Tazelaar, H. D.; McGregor, C. G A.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

Severson, SR, Jeppsson, A, O'Brien, T, Pellegrini, C, Miller, VM, Tazelaar, HD & McGregor, CGA 1998, 'In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs', FASEB Journal, vol. 12, no. 5.
Severson SR, Jeppsson A, O'Brien T, Pellegrini C, Miller VM, Tazelaar HD et al. In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs. FASEB Journal. 1998 Mar 20;12(5).
Severson, S. R. ; Jeppsson, A. ; O'Brien, T. ; Pellegrini, C. ; Miller, Virginia M ; Tazelaar, H. D. ; McGregor, C. G A. / In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs. In: FASEB Journal. 1998 ; Vol. 12, No. 5.
@article{499a12054bfa468bb60665a7fb643e87,
title = "In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs",
abstract = "Experiments were designed to study functional markers for endothelial constitutive nitric oxide synthase (ecNOS) after transbronchial adenoviral-mediated (AD) transfer to transplanted lungs. Syngenic orthotopic single lung transplantation in rats was performed after airway administration of either AdecNOS or the marker gene AdLacZ to the donor lung (300 ul, 1x109 pfu/ml; n=4 each). Transplanted lungs were harvested at four days and transgene DNA and mRNA were identified by polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), respectively. Calcium-dependent NOS activity increased in ecNOS compared to LacZ transduced lungs. However, cyclic GMP isolated from lung homogenates was the same in both groups. Positive immunohistochemical staining for ecNOS was observed in pneumocytes only in ecNOS transduced lungs. Preliminary data indicates that oxidized products of nitric oxide (NOx) increased in pulmonary venous blood in ecNOS transduced lungs. These results indicate an effective transduction of pneumocytes with ecNOS that does not alter basal cyclic GMP levels, although NOS protein and nitric oxide does increase.",
author = "Severson, {S. R.} and A. Jeppsson and T. O'Brien and C. Pellegrini and Miller, {Virginia M} and Tazelaar, {H. D.} and McGregor, {C. G A}",
year = "1998",
month = "3",
day = "20",
language = "English (US)",
volume = "12",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "5",

}

TY - JOUR

T1 - In vivo assessment of endothelial nitric oxide synthase transduced to transplanted lungs

AU - Severson, S. R.

AU - Jeppsson, A.

AU - O'Brien, T.

AU - Pellegrini, C.

AU - Miller, Virginia M

AU - Tazelaar, H. D.

AU - McGregor, C. G A

PY - 1998/3/20

Y1 - 1998/3/20

N2 - Experiments were designed to study functional markers for endothelial constitutive nitric oxide synthase (ecNOS) after transbronchial adenoviral-mediated (AD) transfer to transplanted lungs. Syngenic orthotopic single lung transplantation in rats was performed after airway administration of either AdecNOS or the marker gene AdLacZ to the donor lung (300 ul, 1x109 pfu/ml; n=4 each). Transplanted lungs were harvested at four days and transgene DNA and mRNA were identified by polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), respectively. Calcium-dependent NOS activity increased in ecNOS compared to LacZ transduced lungs. However, cyclic GMP isolated from lung homogenates was the same in both groups. Positive immunohistochemical staining for ecNOS was observed in pneumocytes only in ecNOS transduced lungs. Preliminary data indicates that oxidized products of nitric oxide (NOx) increased in pulmonary venous blood in ecNOS transduced lungs. These results indicate an effective transduction of pneumocytes with ecNOS that does not alter basal cyclic GMP levels, although NOS protein and nitric oxide does increase.

AB - Experiments were designed to study functional markers for endothelial constitutive nitric oxide synthase (ecNOS) after transbronchial adenoviral-mediated (AD) transfer to transplanted lungs. Syngenic orthotopic single lung transplantation in rats was performed after airway administration of either AdecNOS or the marker gene AdLacZ to the donor lung (300 ul, 1x109 pfu/ml; n=4 each). Transplanted lungs were harvested at four days and transgene DNA and mRNA were identified by polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR), respectively. Calcium-dependent NOS activity increased in ecNOS compared to LacZ transduced lungs. However, cyclic GMP isolated from lung homogenates was the same in both groups. Positive immunohistochemical staining for ecNOS was observed in pneumocytes only in ecNOS transduced lungs. Preliminary data indicates that oxidized products of nitric oxide (NOx) increased in pulmonary venous blood in ecNOS transduced lungs. These results indicate an effective transduction of pneumocytes with ecNOS that does not alter basal cyclic GMP levels, although NOS protein and nitric oxide does increase.

UR - http://www.scopus.com/inward/record.url?scp=33749355739&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749355739&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:33749355739

VL - 12

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 5

ER -