Developmental differences in the shear stress-induced expression of endothelial NO synthase: Changing role of AP-1

Stephen Wedgwood, Calista J. Mitchell, Jeffrey R. Fineman, Stephen M. Black

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Endothelial nitric oxide synthase (eNOS) mRNA and protein levels increase during late gestation and then decrease postnatally in sheep lung parenchyma. The increase in fluid shear stress at birth, resulting from increased pulmonary blood flow, is an important mediator of postnatal eNOS gene expression. Our objective was to identify factors stimulating eNOS expression in pulmonary arterial endothelial cells (PAEC) in response to shear stress and to determine if these factors are developmentally regulated. PAEC were isolated from fetal lambs and adult sheep. Transcriptional activity from a 1,600-bp eNOS promoter fragment increased in both fetal and adult PAEC exposed to 8 h of shear stress. Conversely, activity driven from an 840-bp promoter fragment containing a putative activator protein (AP)-1 binding site was increased only in fetal PAEC. This increase was completely abolished in an identical construct containing a mutant AP-1 sequence. The AP-1 protein c-Jun was localized to the cytosol in static adult PAEC and to the nucleus in static fetal PAEC. After shear, c-Jun was nuclear localized in both cell types. However, transcriptionally active phosphorylated c-Jun was elevated only in the nuclei of sheared fetal PAEC. Resting levels of eNOS and NO were 2- and 20-fold higher, respectively, in fetal cells. Shear increased eNOS and NO in both cell types: levels were -2.5-fold higher in fetal PAEC. Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC. We have therefore identified mechanisms of eNOS regulation at the transcriptional level and to be enzyme activation specific to the fetal pulmonary arterial circulation.

Original languageEnglish (US)
Pages (from-to)L650-L662
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume284
Issue number4 28-4
StatePublished - Apr 1 2003

Fingerprint

Transcription Factor AP-1
Nitric Oxide Synthase
Nitric Oxide Synthase Type III
Endothelial Cells
Lung
Sheep
Proto-Oncogene Proteins c-jun
Enzyme Activation
Pulmonary Circulation
Mutant Proteins
Cell Nucleus
Protein Binding
Cytosol
Binding Sites
Phosphorylation
Parturition
Gene Expression
Pregnancy
Messenger RNA

Keywords

  • Activator protein-1
  • Fetal sheep
  • Gene expression
  • Maturational differences
  • Nitric oxide
  • Protein translocation
  • Pulmonary arterial endothelial cells

ASJC Scopus subject areas

  • Physiology
  • Pulmonary and Respiratory Medicine
  • Physiology (medical)
  • Cell Biology

Cite this

Developmental differences in the shear stress-induced expression of endothelial NO synthase : Changing role of AP-1. / Wedgwood, Stephen; Mitchell, Calista J.; Fineman, Jeffrey R.; Black, Stephen M.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 284, No. 4 28-4, 01.04.2003, p. L650-L662.

Research output: Contribution to journalArticle

Wedgwood, Stephen ; Mitchell, Calista J. ; Fineman, Jeffrey R. ; Black, Stephen M. / Developmental differences in the shear stress-induced expression of endothelial NO synthase : Changing role of AP-1. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2003 ; Vol. 284, No. 4 28-4. pp. L650-L662.
@article{94a3faee0093416a80e5985283db1362,
title = "Developmental differences in the shear stress-induced expression of endothelial NO synthase: Changing role of AP-1",
abstract = "Endothelial nitric oxide synthase (eNOS) mRNA and protein levels increase during late gestation and then decrease postnatally in sheep lung parenchyma. The increase in fluid shear stress at birth, resulting from increased pulmonary blood flow, is an important mediator of postnatal eNOS gene expression. Our objective was to identify factors stimulating eNOS expression in pulmonary arterial endothelial cells (PAEC) in response to shear stress and to determine if these factors are developmentally regulated. PAEC were isolated from fetal lambs and adult sheep. Transcriptional activity from a 1,600-bp eNOS promoter fragment increased in both fetal and adult PAEC exposed to 8 h of shear stress. Conversely, activity driven from an 840-bp promoter fragment containing a putative activator protein (AP)-1 binding site was increased only in fetal PAEC. This increase was completely abolished in an identical construct containing a mutant AP-1 sequence. The AP-1 protein c-Jun was localized to the cytosol in static adult PAEC and to the nucleus in static fetal PAEC. After shear, c-Jun was nuclear localized in both cell types. However, transcriptionally active phosphorylated c-Jun was elevated only in the nuclei of sheared fetal PAEC. Resting levels of eNOS and NO were 2- and 20-fold higher, respectively, in fetal cells. Shear increased eNOS and NO in both cell types: levels were -2.5-fold higher in fetal PAEC. Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC. We have therefore identified mechanisms of eNOS regulation at the transcriptional level and to be enzyme activation specific to the fetal pulmonary arterial circulation.",
keywords = "Activator protein-1, Fetal sheep, Gene expression, Maturational differences, Nitric oxide, Protein translocation, Pulmonary arterial endothelial cells",
author = "Stephen Wedgwood and Mitchell, {Calista J.} and Fineman, {Jeffrey R.} and Black, {Stephen M.}",
year = "2003",
month = "4",
day = "1",
language = "English (US)",
volume = "284",
pages = "L650--L662",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "4 28-4",

}

TY - JOUR

T1 - Developmental differences in the shear stress-induced expression of endothelial NO synthase

T2 - Changing role of AP-1

AU - Wedgwood, Stephen

AU - Mitchell, Calista J.

AU - Fineman, Jeffrey R.

AU - Black, Stephen M.

PY - 2003/4/1

Y1 - 2003/4/1

N2 - Endothelial nitric oxide synthase (eNOS) mRNA and protein levels increase during late gestation and then decrease postnatally in sheep lung parenchyma. The increase in fluid shear stress at birth, resulting from increased pulmonary blood flow, is an important mediator of postnatal eNOS gene expression. Our objective was to identify factors stimulating eNOS expression in pulmonary arterial endothelial cells (PAEC) in response to shear stress and to determine if these factors are developmentally regulated. PAEC were isolated from fetal lambs and adult sheep. Transcriptional activity from a 1,600-bp eNOS promoter fragment increased in both fetal and adult PAEC exposed to 8 h of shear stress. Conversely, activity driven from an 840-bp promoter fragment containing a putative activator protein (AP)-1 binding site was increased only in fetal PAEC. This increase was completely abolished in an identical construct containing a mutant AP-1 sequence. The AP-1 protein c-Jun was localized to the cytosol in static adult PAEC and to the nucleus in static fetal PAEC. After shear, c-Jun was nuclear localized in both cell types. However, transcriptionally active phosphorylated c-Jun was elevated only in the nuclei of sheared fetal PAEC. Resting levels of eNOS and NO were 2- and 20-fold higher, respectively, in fetal cells. Shear increased eNOS and NO in both cell types: levels were -2.5-fold higher in fetal PAEC. Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC. We have therefore identified mechanisms of eNOS regulation at the transcriptional level and to be enzyme activation specific to the fetal pulmonary arterial circulation.

AB - Endothelial nitric oxide synthase (eNOS) mRNA and protein levels increase during late gestation and then decrease postnatally in sheep lung parenchyma. The increase in fluid shear stress at birth, resulting from increased pulmonary blood flow, is an important mediator of postnatal eNOS gene expression. Our objective was to identify factors stimulating eNOS expression in pulmonary arterial endothelial cells (PAEC) in response to shear stress and to determine if these factors are developmentally regulated. PAEC were isolated from fetal lambs and adult sheep. Transcriptional activity from a 1,600-bp eNOS promoter fragment increased in both fetal and adult PAEC exposed to 8 h of shear stress. Conversely, activity driven from an 840-bp promoter fragment containing a putative activator protein (AP)-1 binding site was increased only in fetal PAEC. This increase was completely abolished in an identical construct containing a mutant AP-1 sequence. The AP-1 protein c-Jun was localized to the cytosol in static adult PAEC and to the nucleus in static fetal PAEC. After shear, c-Jun was nuclear localized in both cell types. However, transcriptionally active phosphorylated c-Jun was elevated only in the nuclei of sheared fetal PAEC. Resting levels of eNOS and NO were 2- and 20-fold higher, respectively, in fetal cells. Shear increased eNOS and NO in both cell types: levels were -2.5-fold higher in fetal PAEC. Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC. We have therefore identified mechanisms of eNOS regulation at the transcriptional level and to be enzyme activation specific to the fetal pulmonary arterial circulation.

KW - Activator protein-1

KW - Fetal sheep

KW - Gene expression

KW - Maturational differences

KW - Nitric oxide

KW - Protein translocation

KW - Pulmonary arterial endothelial cells

UR - http://www.scopus.com/inward/record.url?scp=0037377477&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037377477&partnerID=8YFLogxK

M3 - Article

C2 - 12533439

AN - SCOPUS:0037377477

VL - 284

SP - L650-L662

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 4 28-4

ER -