Chronic hypoxia decreases endothelial nitric oxide synthase protein expression in fetal guinea pig hearts.
OBJECTIVE
The underlying cellular mechanisms mediating hypoxia-induced adaptations in the fetus are poorly understood. We tested the hypothesis that hypoxia up-regulates endothelial nitric oxide synthase (NOS3, type III) protein expression in fetal hearts similar to that observed in adult hearts as a cardioprotective adaptation. This study investigates the effect of chronic hypoxia on NOS3 protein expression in hearts and carotid arteries of fetal guinea pigs exposed to normoxia or intrauterine hypoxia.


METHODS
Time-mated pregnant guinea pigs (term = 65 days) were housed in either normoxic room air (NMX) or exposed to 12% O(2) (hypoxia; HPX) for 14 or 28 days of duration. At near term ( approximately 60 days of gestation), pregnant mothers were anesthetized and fetal guinea pig hearts and carotid arteries were excised from NMX and HPX animals and frozen until ready for study. In addition, hearts were also excised from anesthetized adult nonpregnant female guinea pigs exposed to either NMX or HPX for 14 days. NOS3 protein was extracted from all tissues and quantified using Western blot analysis. Fetal heart samples were also prepared for localization of NOS3 protein using immunohistochemistry.


RESULTS
Chronic hypoxia increased both maternal and fetal hematocrit after 28 days of duration. HPX decreased NOS3 protein levels in fetal guinea pig hearts by 29% after 28 days compared to NMX controls. In contrast, HPX increased both NOS3 protein levels in adult hearts by 62% and fetal carotid arteries by fourfold after 14 days of exposure compared to their respective NMX controls. Positive immunostaining of NOS3 protein of fetal hearts was localized in both cardiomyocytes and endothelial cells.


CONCLUSIONS
Contrary to our hypothesis, the hypoxia-induced decrease in fetal guinea pig heart NOS3 protein contrasts to the protein levels measured in either adult hearts or fetal carotid arteries. These results suggest that the NOS protein expression is altered differently by hypoxia in fetal and adult hearts and in a peripheral fetal artery exposed to the same level of hypoxia. Thus, the functional role of NO in the fetal heart during hypoxia may differ from that of the adult.
Thompson
Loren P
LP
Department of Obstetrics, Gynecology and Reproductive Sciences, University of Maryland School of Medicine, Baltimore 21201, USA. lthompsonl@umn.edu
Dong
Yafeng
Y
D000818:Animals; D001158:Arteries / Q000201:enzymology; D005260:Female; D047109:Fetal Development; D005311:Fetal Hypoxia; D020869:Gene Expression Profiling; D006168:Guinea Pigs; D006321:Heart / Q000196:embryology*; D007150:Immunohistochemistry; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis*; D011247:Pregnancy; D015854:Up-Regulation
D052250:Nitric Oxide Synthase Type III