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Effect of hyperbaric oxygen (HBO2) treatment on the expression of neuronal nitric oxide synthase (nNOS) in the mouse brain

Treatment with hyperbaric oxygen (HBO2) has been demonstrated by our laboratory and others (Thom and Buerk, Adv. Exp. Med. Biol. 510:133, 2003; Ohgami et al., Brain Res. Bull. 75:668, 2008) to stimulate the production of nitric oxide (NO). The aim of this study was to determine whether exposure to HBO2 increases expression of nNOS enzyme in different regions of the mouse brain. Male NIH Swiss mice were exposed to HBO2 at 3.5 ATA for 60 min then killed at 2, 4, 6 and 8 hr post‐HBO2. The brains were removed and sectioned using a mouse brain matrix to make six 2 mm‐thick coronal sections. Extracts of each section were analyzed by immunoblotting detection of nNOS. Quantification of nNOS expression levels was achieved by measuring the near infrared fluorescence detection of nNOS protein bands. The results demonstrate that the nNOS levels of HBO2‐treated mice were higher than that of room air‐exposed control mice, especially in more caudal regions of the brain, and peaked at 4 hrs post‐HBO2. At 8 hr post‐HBO2, levels of nNOS expression were comparable to that of control mice. Based on these results, we conclude that HBO2 treatment increases the expression of nNOS in the mouse brain. (This research was supported by NIH Grant GM‐77153, the Allen I White Distinguished Professorship and an Institutional SURF Award from ASPET.)