De novo formation of synaptic vesicles was studied electron microscopically in synapses of the superior cervical ganglion of cat. Following prolonged electrical stimulation in a choline deficient condition, almost complete loss of vesicle content was observed. After choline administration, together with the partial restoration of transmission, the reappearance of vesicles could be seen in nerve terminals. In interpreting our observations, the double role of choline (transmitter precursor and membrane constituent) is emphasized.