Individual median octopamine neurons in the insect central nervous system serve as an excellent model system for comparative neuroanatomy of single identified cells. The median octopamine cluster of the subesophageal ganglion consists of defined sets of paired and unpaired interneurons, which supply the brain and subesophageal ganglion with extensive ramifications. The developmental program underlying the complex cellular network is unknown. Here we map the segmental location and developmental origins of individual octopamine neurons in the Drosophila subesophageal ganglion. We demonstrate that two sets of unpaired median neurons, located in the mandibular and maxillary segments, exhibit the same projection patterns in the brain. Furthermore, we show that the paired and unpaired neurons belong to distinct lineages. Interspecies comparison of median neurons revealed that many individual octopamine neurons in different species project to equivalent target regions. Such identified neurons with similar morphology can derive from distinct lineages in different species (i.e., paired and unpaired neurons).