It has been proposed that nitric oxide (NO) is an inhibitory modulator of carotid body (CB) chemoreception to hypoxia. However, the effects of NO gas on carotid chemoreception have not been tested yet. The role played by NO has been revealed by the use of pharmacological tools (i.e., NO donors and NO synthase inhibitors). Here, we studied the effects of NO gas (25 ppm in N(2)) on the chemosensory response to hypoxia (PO(2) approximately 30 Torr) in the cat CB perfused in vitro. During steady hypoxic chemoreceptor excitation, bolus injections or perfusion of Tyrode equilibrated with NO reduced the increased frequency of carotid chemosensory discharges (f(x)). Perfusion for 2 min of Tyrode equilibrated with NO also reduced the rate of the rise of the chemosensory response, as well as the maximal amplitude, as compared with the normal chemosensory response to hypoxia. Present results provide direct evidence that NO gas is an inhibitory modulator of CB hypoxic chemoreception.