Microinjections (100 nL) of 0.15, 0.31, 0.62 and 1.25 mmol/L of nociceptin into the mNTS elicited decreases in mean arterial pressure (MAP; 11 ±1.8, 20 ±2.1, 21.5 ± 3.1 and 15.5 ± 1.9 mmHg, respectively) and heart rate (14 ± 2.7, 29 ±5.5, 39 ±5.2 and 17.5 ± 3.1 beats/min, respectively). Since maximal responses were elicited by microinjections of 0.62 mmol/L of nociceptin, this concentration was used for other experiments. Repeated microinjections of nociceptin (0.62 mmol/L) into the mNTS, at 20-min intervals , did not elicit tachyphylaxis. Bradycardia induced by microinjections of nociceptin into the mNTS was abolished by bilateral vagotomy. The decreases in MAP and HR elicited by nociceptin into the mNTS were blocked by prior microinjections of a specific ORL1 receptor antagonist, [N-phe1]-nociceptin-(1-13)-NH2 (9 mmol/L). Microinjections of the ORL1 receptor antagonist alone did not elicit a response. Prior microinjections of GABA_A and GABA_B receptor antagonists (gabazine, 2 mmol/L and 2-hydroxysaclofen, 100 mmol/L, respectively) into the mNTS, blocked the responses to microinjections of nociceptin at the same site. Prior microinjections of ionotropic glutamate receptor antagonists (NBQX, 2 mmol/L, and D-AP7, 5 mmol/L) also blocked responses to nociceptin microinjections into the mNTS. These results were confirmed by direct neuronal recordings. It was concluded that: 1) nociceptin inhibits GABAergic neurons in the mNTS, 2) GABAergic neurons may normally inhibit the release of glutamate from the terminals of peripheral afferents in the mNTS, 3) Inhibition of GABAergic neurons by nociceptin results in an increase in the release of glutamate in the mNTS, which in turn, elicits depressor and bradycardic responses via activation of ionotropic glutamate receptors on secondary mNTS neurons.