The German cockroach, Blattella germanica, and the double-striped cockroach, B. bisignata, are sibling species with a similar period sequence but a distinctive circadian rhythm in locomotion. The cell distribution of immunoreactivity (ir) against three clock-related proteins, Period (PER), Pigment Dispersing Factor (PDF), and Corazonin (CRZ), was compared between the species. The PER-ir cells tend to form clusters and are sprayed out in the central nervous system. Three major PER-ir cells are located in the optic lobes, which are the sites of the major circadian clock. They are interconnected with PER-ir axon bundles. Interestingly, the potential output signal of the circadian clock, PDF, is co-localized with PER in all three groups of cells. However, only two CRZ-ir cells and their axons are found in the optic lobes and they are not co-localized with PER-ir or PDF-ir cells and axons. Since only one circadian rhythm is expressed in locomotion, the time signals from both major clocks in optic lobes are coupled by connection with PDF-ir axons. A group of 3-4 PER-ir cells in the protocerebrum display typical characteristics of neurosecretary cells. In addition, there are numerous, small PER-ir and PDF-ir co-localized cells in the pars intercerebralis (PI), which have direct connections with the neurohemoorgan, corpora cardiaca, through PER-ir and PDF-ir axons. Based on these findings, the cellular connection shows a circadian control through the endocrine route. For the rest of central nervous system, only a few PER-ir and PDF-ir cells or axons are detected. This finding implies the circadian clock for locomotion is not located in subesophageal ganglion, thoracic or abdominal ganglia, but may use other neural messengers to pass on circadian signals. Since the overall distribution pattern of the clock cells are the same for B. germanica and B. bisignata, the possible explanation for the different expressions of locomotion between the species depends on genes downstream of per, pdf, and crz.
INTRODUCTIONLocating the pacemaker is the first step toward understanding the regulation of circadian systems. By using ablation, the optic lobes were identified as the putative site of the circadian pacemaker in controlling locomotion in cockroaches (Leucophaea maderae, Periplaneta americana, and Blattella germanica) (Nishiitsutsuji-Uwo and Pittendrigh, 1968;Colwell and Page, 1992;Wen and Lee, 2000). The pacemaker was localized between the lobula and the medulla (Helfrich-Förster et al., 1998). Due to technical limitations, however, ablation can not reveal the exact location at the cellular level of the pacemaker. Imunohistochemical analysis, therefore, should provide accuracy to pinpoint the cellular network of circadian clocks, if the essential clock genes are known.The cellular mechanisms of circadian clock involve several feedback loops in the clock cells. The period (per) gene is the main element of the core negative feedback loop in insect and other vertebrate species (Dunlap, 1999;Hardin, 2000). In the Drosophila model (...