Upregulation of nitric oxide synthase in mice with severe hypoxia-induced pulmonary hypertension.
BACKGROUND
The importance of nitric oxide (NO) in hypoxic pulmonary hypertension has been demonstrated using nitric oxide synthase (NOS) knockout mice. In that model NO from endothelial NOS (eNOS) plays a central role in modulating pulmonary vascular tone and attenuating hypoxic pulmonary hypertension. However, the normal regulation of NOS expression in mice following hypoxia is uncertain. Because genetically engineered mice are often utilized in studies of NO, we conducted the present study to determine how hypoxia alters NOS expression in wild-type mice.


METHODS
Mice were exposed to sea level, ambient conditions (5280 feet) or severe altitude (17,000 feet) for 6 weeks from birth, and hemodynamics and lung NOS expression were assessed.


RESULTS
Hypoxic mice developed severe pulmonary hypertension (right ventricular systolic pressure [RVsP] 60 mmHg) as compared with normoxic mice (27 mmHg). Using quantitative reverse-transcription PCR, it was found that expressions of eNOS and inducible NOS (iNOS) increased 1.5-fold and 3.5-fold, respectively, in the lung. In addition, the level of lung eNOS protein was increased, neuronal NOS (nNOS) protein was unchanged, and iNOS was below the limit of detection. Immunohistochemistry demonstrated no change in lung iNOS or nNOS staining in either central or peripheral areas, but suggested increased eNOS in the periphery following hypoxia.


CONCLUSIONS
In mice, hypoxia is associated with increases in lung eNOS, possibly in iNOS, but not in nNOS; this suggests that the pattern of lung NOS expression following hypoxia must be considered in studies using genetically engineered mice.
Fagan
K A
KA
Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Health Sciences Center, 4200 East Ninth Avenue B-133, Denver, CO 80262, USA. karen.fagan@uchsc.edu
Morrissey
B
B
Fouty
B W
BW
Sato
K
K
Harral
J W
JW
Morris
K G
KG
Hoedt-Miller
M
M
Vidmar
S
S
McMurtry
I F
IF
Rodman
D M
DM
D000818:Animals; D001794:Blood Pressure / Q000502:physiology; D015153:Blotting, Western; D006400:Hematocrit; D006976:Hypertension, Pulmonary / Q000201:enzymology* / Q000209:etiology / Q000473:pathology; D000860:Hypoxia / Q000150:complications / Q000201:enzymology* / Q000473:pathology; D007150:Immunohistochemistry; D008168:Lung / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000502:physiology*; D016278:Ventricular Function, Right
D012333:RNA, Messenger; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase