1934 Feldberg and Vartiainen et al claimed that Acetylcholine liberates only at the synapses by the passing of nervous impulses. However recently Lorent du No reported that such a release took place not only when the sympathetic preganglionic fibres were stimulated, but also when vagal or sympathetic postganglionic elements were activated. This problem has not yet been ascertaind with certainty. The purpose of the present investigation is to elucidate the site of the liberation of Acetylcholine. Superior cervical ganglion, ganglion nodosum of vagus and spinal ganglia and also ganglia of sympathetic chain of rabbits were stimulated electrically some minutes long in vivo from their nerve trunks and Acetylcholine content or these ganglia was compared with unstimulated. Among the ganglia, which I stimulated include two types of ganglion i.e. ganglion with synapses and without synapses. Acetylcholine content of all these ganglia increased in fair amounts when stimulated. From these results we can conclude as follows; the liberation of Acetylcholine is a process that is not specific to the synapses and the nerve endigs, but also liberate from ganglion without synapses.