Nitric oxide is used as an orthograde cotransmitter at identified histaminergic synapses.

1. Nitric oxide (NO) is produced by the enzyme nitric oxide synthase (NOS) and serves as an interneuronal messenger. Here, an identified histaminergic neuron (C2) in the mollusk Aplysia californica is shown to contain NOS with the use of NADPH diaphorase staining, suggesting that NO and histamine are used as cotransmitters by neuron C2. 2. The NOS containing neuron, C2, evokes a very slow excitatory postsynaptic potential in two of its identified postsynaptic follower neurons that are insensitive to H1 and H2 histamine receptor antagonists. 3. The very slow excitatory postsynaptic potential is blocked by inhibitors of NOS, such as nitroarginine methyl ester, and suppressed by the NO scavenger reduced hemoglobin. 4. Treatments with compounds that release NO, such as nitrosocysteine, mimic the membrane depolarization and the decrease in membrane conductance in the follower that are characteristic of the very slow excitatory postsynaptic potential induced normally by the presynaptic C2 neuron. 5. These results indicate that NO is used as an orthograde synaptic cotransmitter at synapses between histaminergic neuron C2 and its followers that receive the very slow excitatory postsynaptic potential.