Evidence for a role of orcokinin-related peptides in the circadian clock controlling locomotor activity of the cockroach Leucophaea maderae.

The accessory medulla (AMe), a small neuropil in the optic lobe, houses the master circadian clock in the brain of the cockroach Leucophaea maderae and controls circadian rhythms in locomotor activity. Recently, members of the orcokinin family of crustacean neuropeptides were identified in a cockroach and a locust and were shown by immunocytochemistry to be prominently present in the AMe. In the cockroach L. maderae, about 30 neurons in five of six established cell groups of the AMe showed orcokinin immunostaining. By means of tracer injections into one AMe and immunostaining with anti-orcokinin antiserum, we show here that one orcokinin-immunoreactive ventral neuron and three ventromedian neurons directly connect both AMae. To determine a possible circadian function of orcokinin in the cockroach, we injected 150 fmol Asn(13)-orcokinin into the vicinity of the AMe at different circadian times. These experiments resulted in stable phase-dependent phase shifts of circadian locomotor activity of the cockroach. The shape of the resulting phase-response curve closely matched the phase-shifting effects of light pulses, and its amplitude was dependent on the amount of the injected peptide. Together with the anatomical data, the results suggest that orcokinin-related peptides play an important role in light entrainment pathways to the circadian clock via the contralateral compound eye. This study, furthermore, provides the first evidence for a physiological role of an orcokinin-related peptide in insects.