title: In vivo expression profile of an endothelial nitric oxide synthase promoter-reporter transgene.
abstract: 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 beta-galactosidase. Examination of beta-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 beta-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.
authors.affiliation: Department of Medicine, St. Michael's Hospital and University of Toronto, Toronto M5S 1A8, Ontario, Canada M5S 1X8.
authors.forename: A M
authors.initials: AM
authors.lastname: Teichert
authors.forename: T L
authors.initials: TL
authors.lastname: Miller
authors.forename: S C
authors.initials: SC
authors.lastname: Tai
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: X
authors.initials: X
authors.lastname: Bei
authors.forename: G B
authors.initials: GB
authors.lastname: Robb
authors.forename: M J
authors.initials: MJ
authors.lastname: Phillips
authors.forename: P A
authors.initials: PA
authors.lastname: Marsden
mesh_terms: D000818:Animals; D001161:Arteriosclerosis / Q000201:enzymology; D004730:Endothelium, Vascular / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D006973:Hypertension / Q000201:enzymology; D007606:Juxtaglomerular Apparatus / Q000201:enzymology; D007668:Kidney / Q000098:blood supply / Q000502:physiology; D007763:Lac Operon; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016254:Mutagenesis, Insertional; D009399:Nephrons / Q000201:enzymology; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012079:Renal Circulation / Q000502:physiology; D019076:Transgenes / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse