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1 Department of Medicine and 3 Renal Division, St. Michael's Hospital and University of Toronto, Toronto M5S 1A8; and 2 Department of Pathology, Hospital for Sick Children, Toronto, Ontario, Canada M5S 1X8
Endothelium-derived nitric oxide (NO) is primarily
attributable to constitutive expression of the endothelial nitric oxide synthase (eNOS) gene. Although a more comprehensive understanding of
transcriptional regulation of eNOS is emerging with respect to in vitro
regulatory pathways, their relevance in vivo warrants assessment. In
this regard, promoter-reporter insertional transgenic murine lines were
created containing 5,200 bp of the native murine eNOS promoter
directing transcription of nuclear-localized 
-galactosidase. Examination of -galactosidase expression in heart, lung, kidney, liver, spleen, and brain of adult mice demonstrated robust signal in
large and medium-sized blood vessels. Small arterioles, capillaries, and venules of the microvasculature were notably negative, with the
exception of the vasa recta of the medullary circulation of the kidney,
which was strongly positive. Only in the brain was the reporter
expressed in non-endothelial cell types, such as the CA1 region of the
hippocampus. Epithelial cells of the bronchi, bronchioles, and alveoli
were scored as negative, as was renal tubular epithelium. Cardiac
myocytes, skeletal muscle, and smooth muscle of both vascular and
nonvascular sources failed to demonstrate -galactosidase staining.
Expression was uniform across multiple founders and was not
significantly affected by genomic integration site. These transgenic
eNOS promoter-reporter lines will be a valuable resource for ongoing
studies addressing the regulated expression of eNOS in vivo in both
health and disease.
atherosclerosis; endothelium; hypertension; gene regulation
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