Cytosolic and membrane-bound cerebral nitric oxide synthase activity during hypoxia in cortical tissue of newborn piglets

To determine the role of nitric oxide production during hypoxia, the presence of two forms of neuronal nitric oxide synthase, cytosolic (cNOS) and membrane-bound (memNOS), in cortical tissue of newborn piglets and the effects of hypoxia on the activity of these enzymes were studied. Experiments were performed in 12 anesthetized and ventilated Yorkshire piglets, 2-4 days of age. Hypoxia was induced by decreasing the FiO(2) to 0.07. The control group was ventilated maintaining normoxia. After 1 h of normoxic or hypoxic ventilation brain tissue was removed and frozen immediately in liquid nitrogen. Tissue hypoxia was confirmed by analysis of adenosine triphosphate (ATP) and phosphocreatine (PCr): ATP was reduced to 52% and PCr to 28% of control values, cNOS activity was 35.3 +/- 13.7 pmol/mg protein per min in the control group and 28.3 +/- 7.0 in the hypoxia group; memNOS activity was 10.5 +/- 4.5 and 12.0 +/- 3.9 pmol/mg protein per min in the control and hypoxia groups, respectively. Differences in cNOS and memNOS activity between control and hypoxic animals were not significant. The results indicate that both cNOS and memNOS are present in cortical tissue of newborn piglets and that the activity is unaffected by 1 h of tissue hypoxia. We suggest that production of nitric oxide and its derivative peroxynitrite during hypoxia may therefore be a potential mechanism for hypoxia-induced brain cell membrane lipid peroxidation.