3D-atlas of the brain of the cockroach Rhyparobia maderae.

The Madeira cockroach Rhyparobia maderae is a nocturnal insect and a prominent model organism for the study of circadian rhythms. Its master circadian clock, controlling circadian locomotor activity and sleep-wake cycles, is located in the accessory medulla of the optic lobe. For a better understanding of brain regions controlled by the circadian clock and brain organization of this insect in general, we created a three-dimensional (3D) reconstruction of all neuropils of the cerebral ganglia based on anti-synapsin and anti-γ-aminobutyric acid immunolabeling of whole mount brains. Forty-nine major neuropils were identified and three-dimensionally reconstructed. Single-cell dye fills complement the data and provide evidence for distinct subdivisions of certain brain areas. Most neuropils defined in the fruit fly Drosophila melanogaster could be distinguished in the cockroach as well. However, some neuropils identified in the fruit fly do not exist as distinct entities in the cockroach while others are lacking in the fruit fly. In addition to neuropils, major fiber systems, tracts, and commissures were reconstructed and served as important landmarks separating brain areas. Being a nocturnal insect, R. maderae is an important new species to the growing collection of 3D insect brain atlases and only the second hemimetabolous insect, for which a detailed 3D brain atlas is available. This atlas will be highly valuable for an evolutionary comparison of insect brain organization and will greatly facilitate addressing brain areas that are supervised by the circadian clock.