S U M M A R Y 1. The actions have been compared of the acetylcholine antagonists, hexamethonium, tuborcurarine, pancuronium and gallamine on the effects of neostigmine at the neuromuscular junction of the soleus muscle in cats under chloralose anaesthesia.2. Neostigmine produced augmentation of twitch tension, muscle fasciculat ions, repetitive firing in the muscle and antidromic repetitive firing in the ventral roots: all these effects were blocked by the acetylcholine antagonists. The effective doses of hexamethonium were similar to its ganglion blocking doses, being much smaller than those necessary to block the normal soleus twitch, and it is concluded i.hat the effects of neostigmine arise mainly from the action of preserved transmitter (acetylcholine) on the motor nerve endings.3. In the smallest effective doses, hexamethonium preferentially blocked muscle fasciculations and the associated electrical activity in nerve. Tubocurarine, which has considerable ganglion blocking activity (reflecting its affinity for neuronal cholinoceptors) resembled hexamethonium. In contrast, pancuronium and gallamine, which are considered to have a higher affinity for post-junctional muscle cholinoceptors, depressed muscle fasciculations only in doses that also depressed twitch augmentation and repetitive firing in muscle.4. In slightly larger doses, pancuronium and gallamine also blocked antidromic repetitive firing in nerve indicating that these drugs also combine with prejunctional neuronal cholinoceptors, and suggesting that these cholinoceptors are intermediate in character between ganglionic and muscle cholinoceptors.