Nitric oxide metabolites in goldfish under normoxic and hypoxic conditions.
Nitric oxide (NO), produced by nitric oxide synthases (NOS enzymes), regulates multiple physiological functions in animals. NO exerts its effects by binding to iron (Fe) of heme groups (exemplified by the activation of soluble guanylyl cyclase) and by S-nitrosylation of proteins - and it is metabolized to nitrite and nitrate. Nitrite is used as a marker for NOS activity but it is also a NO donor that can be activated by various cellular proteins under hypoxic conditions. Here, we report the first systematic study of NO metabolites (nitrite, nitrate, S-nitroso, N-nitroso and Fe-nitrosyl compounds) in multiple tissues of a non-mammalian vertebrate (goldfish) under normoxic and hypoxic conditions. NO metabolites were measured in blood (plasma and red cells) and heart, brain, gill, liver, kidney and skeletal muscle, using highly sensitive reductive chemiluminescence. The severity of the chosen hypoxia levels was assessed from metabolic and respiratory variables. In normoxic goldfish, the concentrations of NO metabolites in plasma and tissues were comparable with values reported in mammals, indicative of similar NOS activity. Exposure to hypoxia [at P(O₂) (partial pressure of O₂) values close to and below the critical P(O₂)] for two days caused large decreases in plasma nitrite and nitrate, which suggests reduced NOS activity and increased nitrite/nitrate utilization or loss. Tissue NO metabolites were largely maintained at their tissue-specific values under hypoxia, pointing at nitrite transfer from extracellular to intracellular compartments and cellular NO generation from nitrite. The data highlights the preference of goldfish to defend intracellular NO homeostasis during hypoxia.
Hansen
Marie N
MN
Institute of Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
Jensen
Frank B
FB
D000818:Animals; D004912:Erythrocytes / Q000378:metabolism; D005880:Gills / Q000166:cytology / Q000201:enzymology; D006054:Goldfish / Q000097:blood / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D000860:Hypoxia / Q000097:blood / Q000378:metabolism*; D019344:Lactic Acid / Q000097:blood; D009566:Nitrates / Q000097:blood; D009569:Nitric Oxide / Q000097:blood / Q000378:metabolism*; D009928:Organ Specificity; D000254:Sodium-Potassium-Exchanging ATPase
D006454:Hemoglobins; D009566:Nitrates; D009569:Nitric Oxide; D019344:Lactic Acid; D000254:Sodium-Potassium-Exchanging ATPase