Neurotransmitters play a crucial role in the communication between neurons in the autonomic nervous system. Neurons in the autonomic nervous system can be cholinergic or adrenergic depending on the neurotransmitters synthesized. Cholinergic neurons use acetylcholine as their primary neurotransmitter. This includes all the preganglionic fibers of the sympathetic and pre- and postganglionic fibers of the parasympathetic nervous systems. In addition, neurons of the somatic nervous system also use acetylcholine at the neuromuscular junction. In contrast, adrenergic neurons employ noradrenaline (also known as norepinephrine) as their main neurotransmitter. The postganglionic neurons at the sympathetic division use noradrenaline for transmission. Furthermore, the adrenal medulla releases a mixture of adrenaline and noradrenaline.

Apart from acetylcholine and noradrenaline, nonadrenergic noncholinergic (NANC) transmitters are also involved in autonomic transmission. These include ATP, neuropeptide Y (NPY), vasoactive intestinal peptides (VIP), and nitric oxide, which function at the postganglionic nerve terminal. Neurotransmitters such as γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT), dopamine, and substance P involved in ganglionic transmissions, are also called NANC.

Sometimes, neurotransmitters are often accompanied by co-transmitters during release. Co-transmitter helps the primary neurotransmitter in reaching remote targets and producing sustained effects. For example, acetylcholine is often associated with VIP, while acetylcholine and noradrenaline are associated with ATP. Co-transmitters are stored in separate vesicles from the neurotransmitter. Although, certain co-transmitters, such as ATP, may be stored in the same vesicle as noradrenaline. Co-transmitters can also serve as alternative transmitters, exerting trophic effects on the synapses.