The pars intercerebralis as a modulator of locomotor rhythms and feeding in the American cockroach, Periplaneta americana.

It has been shown that in orthopteran insects each of the optic lobes (OLs) contains a circadian pacemaker controlling locomotor activity and that the pars intercerebralis (PI) modifies the activity level. However, the present study showed Period protein-like immunoreactivity (PER-ir) in the PI and dorsolateral protocerebrum (DL) as well as in the OLs in the American cockroach, Periplaneta americana, which raised the possibility that the PI or DL could be a clock element. Therefore, we removed the PI or DL surgically and observed the effects on locomotor rhythms and feeding behavior. In constant darkness (DD), cockroaches with an ablated PI (PIX-DD) showed arrhythmicity in locomotion and a massive increase in food consumption that led to increased body length and weight, while PIX cockroaches reared under LD 12:12 (PIX-LD) and the sham-treated cockroaches in DD (CNT-DD) showed rhythmicity and no increase in food consumption. Statistical analysis showed that arrhythmicity was not accompanied by hyperactivity, suggesting that the PI is involved in the regulation of locomotor activity and feeding in DD. The activities of alpha-amylase and proteases were found to be markedly elevated in the midgut of PIX-DD cockroaches but not in PIX-LD cockroaches. Taken together, these results indicate that the PI modulates locomotor rhythms and feeding behavior of cockroaches in a light-dependent manner. The PI and the OL may regulate circadian rhythms and feeding via distinct pathways.

Day/night fluctuations in melatonin content, arylalkylamine N-acetyltransferase activity and NAT mRNA expression in the CNS, peripheral tissues and hemolymph of the cockroach, Periplaneta americana.

Melatonin content measured by a radioenzymatic assay in the brain of the American cockroach (Periplaneta americana) showed a day/night fluctuation with higher levels at night under LD 12:12. The activity of arylalkylamine N-acetyltransferase (NAT) in brain was also higher at night and this pattern continued in constant darkness. The results suggest that the rhythmicity in melatonin content can be caused by NAT. Melatonin content in hemolymph showed an even greater day/night difference, more than 12 times that in brain under LD 12:12. Melatonin levels in retina were also higher at night while NAT activity was not significantly higher at night than at daytime. Using a probe designed from NAT cloned from testes we performed Northern blot analysis of total RNA, which revealed that the level of NAT mRNA was higher in midgut, ovary and female accessory glands than in fat body and brain. The level of transcript in midgut was higher at night, but the levels in ovary and female accessory reproductive gland showed the opposite pattern. We also used the antibody to whole Drosophila melanogaster aaNAT1 protein, seeking a homologous antigen in the cephalic ganglia. NAT-like antigen was detected in several restricted populations of cells in the brain that were partially co-localized with PER-like antigen. The results suggest that NAT exists in multiple forms in various tissues of the cockroach and that its functions and regulations can vary among tissues. The results in the brain led to the conclusion that NAT could be a clock-controlled gene functioning as an output regulator of the circadian clock.