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Updated: Aug 30, 2026
Analytical Techniques for Assaying Nitric Oxide Bioactivity
Published on: June 18, 2012
A technique to estimate the rate of whole body nitric oxide formation in conscious mice
Anna Wickman1, Natalia Klintland, Li-ming Gan
1Department of Physiology, Sahlgrenska Academy, Göteborg University, PO Box 432, S-405 30, Göteborg, Sweden. anna.wickman@fysiologi.gu.se
Objective:
We have established a technique to estimate the total rate of nitric oxide (NO) formation in mice, based on inhalation of a stable oxygen isotope (18O(2)). Changes of NO production with age were also studied.
Methods:
The experiments were performed in eight-week- (n=6) and eight-month-old (n=6-7), respectively, female (C57/Bl6xCBAca) mice. Pairs of conscious mice were kept in an air-tight closed system allowing breathing of a mixture containing 18O(2). The 18O(2)-technique was validated by L-NAME (10mg/kg) and lipopolysaccharide (LPS, 8 mg/kg) administration. The concentrations of O(2) and CO(2) in the system were controlled and plasma nitrate analyzed by GC/MS technique.
Results:
NO formation was similar in young and old mice (young=7.68+/-1.47 vs. old = 6.25+/-1.49 micromol/kg/h, n.s.). Total NO production was reduced after L-NAME treatment in young animals by 91% and in old animals by 71% (p<0.05 for both), whereas LPS administration increased NO production (114+/-17%, p<0.05).Conclusion. NO formation is unaltered with age in mice. The 18O(2)-technique is a valid and specific technique to estimate whole body NO production in conscious mice.
