Critical roles of mecamylamine-sensitive mushroom body neurons in insect olfactory learning

In insects, cholinergic neurons are thought to transmit olfactory conditioned stimulus (CS) to the sites for associating the CS with unconditioned stimulus (US), but the types of acetylcholine (ACh) receptor used by neurons participating in the association have not been determined. In cockroaches, a type of nicotinic ACh receptor specifically antagonized by mecamylamine (MEC) has been characterized. Here we investigated the roles of neurons possessing MEC-sensitive ACh receptors (MEC-sensitive neurons) in olfactory conditioning of salivation, monitored by changes in activities of salivary neurons, in cockroaches. Local and bilateral microinjection of MEC into each of the three olfactory centers, antennal lobes, calyces of the mushroom bodies and lateral protocerebra, impaired olfactory responses of salivary neurons, indicating that MEC-sensitive neurons in all olfactory centers participate in pathways mediating olfactory responses of salivary neurons. Conditioning of olfactory CS with sucrose US was impaired by injection of MEC into the antennal lobes or calyces, i.e., conditioned responses were absent even after recovery from MEC injection, suggesting that the CS-US association occurs in MEC-sensitive neurons in calyces (most probably Kenyon cells) or in neurons in downstream pathways. In contrast, conditioned responses appeared after recovery from MEC injection into the lateral protocerebra, suggesting that MEC-sensitive neurons in the lateral protocerebra are downstream of the association sites. Since lateral protocerebra are major termination areas of mushroom body efferent neurons, we suggest that input synapses of MEC-sensitive Kenyon cells, or their output synapses upon mushroom body efferent neurons, are the sites for CS-US association for conditioning of salivation.