Acetylcholine is stored at the end of nerve cells until its triggered to be released. Once released from the end of the nerve cell, it moves into a space called the synaptic cleft. The synaptic cleft is between the nerve cell from which acetylcholine was released (the presynaptic nerve cell) and the next nerve cell acetylcholine is going to (the postsynaptic nerve cell).

Once acetylcholine moves across the synapse, it can bind to two types of receptors: nicotinic receptors and muscarinic receptors. There are two subtypes of nicotinic receptors and five types of muscarinic receptors. After binding to the receptors, the chemical message moves along to the next nerve cell and then the process repeats until the message arrives at its destination.

Acetylcholine in the synapse is broken down by an enzyme called acetylcholinesterase into choline and acetate. These products are reabsorbed and recycled so they can be used again in transmitting another chemical message.