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The clock gene cycle plays an important role in the circadian clock of the cricket Gryllus bimaculatus

•Gb’cyc is rhythmically expressed in light/dark cycles, peaking at mid night, in Gryllus bimaculatus.•cyc RNAi lengthened the free-running period of locomotor rhythms in DD.•cyc RNAi crickets showed a rhythmic expression of Gb’per, Gb’tim and Gb’Clk in the optic lobe in DD.•The cricket might have a...

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Published in:Journal of insect physiology 2013-07, Vol.59 (7), p.697-704
Main Authors: Uryu, Outa, Karpova, Svetlana G., Tomioka, Kenji
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description •Gb’cyc is rhythmically expressed in light/dark cycles, peaking at mid night, in Gryllus bimaculatus.•cyc RNAi lengthened the free-running period of locomotor rhythms in DD.•cyc RNAi crickets showed a rhythmic expression of Gb’per, Gb’tim and Gb’Clk in the optic lobe in DD.•The cricket might have a unique clock with oscillatory mechanisms for rhythmic expression of Gb’cyc and Gb’Clk. To dissect the molecular oscillatory mechanism of the circadian clock in the cricket Gryllus bimaculatus, we have cloned a cDNA of the clock gene cycle (Gb’cyc) and analyzed its structure and function. Gb’cyc contains four functional domains, i.e. bHLH, PAS-A, PAS-B and BCTR domains, and is expressed rhythmically in light dark cycles, peaking at mid night. The RNA interference (RNAi) of Clock (Gb’Clk) and period (Gb’per) reduced the Gb’cyc mRNA levels and abolished the rhythmic expression, suggesting that the rhythmic expression of Gb’cyc is regulated by a mechanism including Gb’Clk and Gb’per. These features are more similar to those of mammalian orthologue of cyc (Bmal1) than those of Drosophila cyc. A single treatment with double-stranded RNA (dsRNA) of Gb’cyc effectively knocked down the Gb’cyc mRNA level and abolished its rhythmic expression. The cyc RNAi failed to disrupt the locomotor rhythm, but lengthened its free-running period in constant darkness (DD). It is thus likely that Gb’cyc is involved in the circadian clock machinery of the cricket. The cyc RNAi crickets showed a rhythmic expression of Gb’per and timeless (Gb’tim) in the optic lobe in DD, explaining the persistence of the locomotor rhythm. Surprisingly, cyc RNAi revealed a rhythmic expression of Gb’Clk in DD which is otherwise rather constitutively expressed in the optic lobe. These facts suggest that the cricket might have a unique clock oscillatory mechanism in which both Gb’cyc and Gb’Clk are rhythmically controlled and that under abundant expression of Gb’cyc the rhythmic expression of Gb’Clk may be concealed.
doi_str_mv 10.1016/j.jinsphys.2013.04.011
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To dissect the molecular oscillatory mechanism of the circadian clock in the cricket Gryllus bimaculatus, we have cloned a cDNA of the clock gene cycle (Gb’cyc) and analyzed its structure and function. Gb’cyc contains four functional domains, i.e. bHLH, PAS-A, PAS-B and BCTR domains, and is expressed rhythmically in light dark cycles, peaking at mid night. The RNA interference (RNAi) of Clock (Gb’Clk) and period (Gb’per) reduced the Gb’cyc mRNA levels and abolished the rhythmic expression, suggesting that the rhythmic expression of Gb’cyc is regulated by a mechanism including Gb’Clk and Gb’per. These features are more similar to those of mammalian orthologue of cyc (Bmal1) than those of Drosophila cyc. A single treatment with double-stranded RNA (dsRNA) of Gb’cyc effectively knocked down the Gb’cyc mRNA level and abolished its rhythmic expression. The cyc RNAi failed to disrupt the locomotor rhythm, but lengthened its free-running period in constant darkness (DD). 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To dissect the molecular oscillatory mechanism of the circadian clock in the cricket Gryllus bimaculatus, we have cloned a cDNA of the clock gene cycle (Gb’cyc) and analyzed its structure and function. Gb’cyc contains four functional domains, i.e. bHLH, PAS-A, PAS-B and BCTR domains, and is expressed rhythmically in light dark cycles, peaking at mid night. The RNA interference (RNAi) of Clock (Gb’Clk) and period (Gb’per) reduced the Gb’cyc mRNA levels and abolished the rhythmic expression, suggesting that the rhythmic expression of Gb’cyc is regulated by a mechanism including Gb’Clk and Gb’per. These features are more similar to those of mammalian orthologue of cyc (Bmal1) than those of Drosophila cyc. A single treatment with double-stranded RNA (dsRNA) of Gb’cyc effectively knocked down the Gb’cyc mRNA level and abolished its rhythmic expression. The cyc RNAi failed to disrupt the locomotor rhythm, but lengthened its free-running period in constant darkness (DD). It is thus likely that Gb’cyc is involved in the circadian clock machinery of the cricket. The cyc RNAi crickets showed a rhythmic expression of Gb’per and timeless (Gb’tim) in the optic lobe in DD, explaining the persistence of the locomotor rhythm. Surprisingly, cyc RNAi revealed a rhythmic expression of Gb’Clk in DD which is otherwise rather constitutively expressed in the optic lobe. These facts suggest that the cricket might have a unique clock oscillatory mechanism in which both Gb’cyc and Gb’Clk are rhythmically controlled and that under abundant expression of Gb’cyc the rhythmic expression of Gb’Clk may be concealed.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>23665334</pmid><doi>10.1016/j.jinsphys.2013.04.011</doi><tpages>8</tpages></addata></record>
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ispartof Journal of insect physiology, 2013-07, Vol.59 (7), p.697-704
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subjects Animals
Circadian clock
Circadian Clocks
circadian rhythm
Clock genes
CLOCK Proteins - genetics
CLOCK Proteins - metabolism
complementary DNA
Cricket
Cycle
double-stranded RNA
Drosophila
gene expression
genes
Gryllidae - classification
Gryllidae - genetics
Gryllidae - physiology
Gryllidae - radiation effects
Gryllus bimaculatus
Insect Proteins - genetics
Insect Proteins - metabolism
mammals
messenger RNA
optic lobe
Photoperiod
Phylogeny
RNA Interference
RNAi
title The clock gene cycle plays an important role in the circadian clock of the cricket Gryllus bimaculatus
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