Molenaar_1983_Neurosci.Lett_43_209

Bound and free acetylcholine (ACh) were measured in frog sartorius muscles by mass fragmentography. Upon incubation of the muscles for 5 min with potassium propionate, which stimulated the release of ACh, there was a 2-fold reduction of bound ACh. In contrast, the amount of free ACh remained unchanged. After 65 min recovery from stimulation in normal Ringer solution containing deuterium-labelled choline, free ACh was labelled to a higher degree than bound ACh. The results are in agreement with the idea that ACh is synthesized in the cytoplasmic compartment of the motor nerve terminal, and subsequently transferred to the vesicles from which it is released upon stimulation.