To investigate whether nitric oxide (NO) in the dorsal motor nucleus of the vagus (DMV) mediated an influence on the extrahepatic biliary system, we studied the effects of microinjection of NO-producing drugs into DMV on the motilities of the gallbladder (GB) and the sphincter of Oddi (SO) in anesthetized rabbits. Microinjection of the NO precursor L-arginine into the rostral DMV produced an increase in the GB and SO motilities, which can be counteracted by both NG-nitro-L-arginine-methyl (L-NAME), an inhibitor of NOS, and reduced hemoglobin (rHb), a scavenger of NO, and were eliminated by bilateral cervical vagotomy. On the other hand, the NO donor sodium nitroprusside (SNP) was able to mimic the excitatory effect of L-arginine. This effect can be antagonized by rHb, but not by L-NAME, for SNP supplied exogenous NO without activating NOS. These results indicate that NO in the DMV mediates an excitatory effect on the extrahepatic biliary system.