Hypoxia induces a functionally significant and translationally efficient neuronal NO synthase mRNA variant.
We tested the hypothesis that induction of neuronal NO synthase (nNOS) impairs vascular smooth muscle contractility after hypoxia. nNOS protein was increased in aorta, mesenteric arterioles, pulmonary arteries, brain, and diaphragm from rats exposed to 8% O2 for 48 hours and in human aortic SMCs after hypoxic incubation (1% O2). Ca-dependent NO synthase activity was increased in endothelium-denuded aortic segments from hypoxia-exposed rats. N-nitro-L-arginine methyl ester enhanced the contractile responses of endothelium-denuded aortic rings and mesenteric arterioles from hypoxia-exposed but not normoxic rats (P < 0.05). The hypoxia-inducible mRNA transcript expressed by human cells was found to contain a novel 5'-untranslated region, consistent with activation of transcription in the genomic region contiguous with exon 2. Translational efficiency of this transcript is markedly increased compared with previously described human nNOS mRNAs. Transgenic mice possessing a lacZ reporter construct under control of these genomic sequences demonstrated expression of the construct after exposure to hypoxia (8% O2, 48 hours) in the aorta, mesenteric arterioles, renal papilla, and brain. These results reveal a novel human nNOS promoter that confers the ability to rapidly upregulate nNOS expression in response to hypoxia with a functionally significant effect on vascular smooth muscle contraction.
Ward
Michael E
ME
Division of Respirology, University of Toronto, Toronto, Ontario, Canada. wardm@smh.toronto.on.ca
Toporsian
Mourad
M
Scott
Jeremy A
JA
Teoh
Hwee
H
Govindaraju
Vasanthi
V
Quan
Adrian
A
Wener
Avraham D
AD
Wang
Guilin
G
Bevan
Siân C
SC
Newton
Derek C
DC
Marsden
Philip A
PA
D000818:Animals; D001013:Aorta, Thoracic / Q000378:metabolism; D015153:Blotting, Western; D017930:Genes, Reporter; D014644:Genetic Variation; D006801:Humans; D000860:Hypoxia / Q000201:enzymology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D052248:Nitric Oxide Synthase Type I / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley
D012333:RNA, Messenger; D052248:Nitric Oxide Synthase Type I