The effect of pulsed magnetic fields on nitric oxide synthase (NOS) activity in the rat brain was investigated. Sprague–Dawley rats (male, 200–250 g body weight) brain were dissected regionally, and the crude enzyme solutions were treated with pulsed DC, AC or static DC magnetic fields at 0°C for 1 h. After exposure, NOS activity was measured as nitrite and nitrate levels generated from incubation with arginine, CaCl 2 and β-nicotinamide adenine dinucleotide phosphate. Under these experimental conditions, neither AC nor static DC field treatment showed any significant change in NOS activity. A significant increase in NOS activity was observed in the cerebellum (111.2±2.0%, P<0.05, five separate experiments) for a 1 Gauss (0.1 mT) pulsed DC field. Under the same experimental condition, only a slight change or no effect was observed in the hippocampus, cortex, medulla oblongata, hypothalamus, striatum and midbrain. These studies suggest that pulsed magnetic fields result in a different effect on NOS activity in the cerebellum of the rats.