Author Correction: The characterization of the circadian clock in the olive fly Bactrocera oleae (Diptera: Tephritidae) reveals a Drosophila-like organization.

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The characterization of the circadian clock in the olive fly Bactrocera oleae (Diptera: Tephritidae) reveals a Drosophila-like organization.

The olive fruit fly, Bactrocera oleae, is the single most important pest for the majority of olive plantations. Oxitec's self-limiting olive fly technology (OX3097D-Bol) offers an alternative management approach to this insect pest. Because of previously reported asynchrony in the mating time of wild and laboratory strains, we have characterized the olive fly circadian clock applying molecular, evolutionary, anatomical and behavioural approaches. Here we demonstrate that the olive fly clock relies on a Drosophila melanogaster-like organization and that OX3097D-Bol carries a functional clock similar to wild-type strains, confirming its suitability for operational use.

A Tug-of-War between Cryptochrome and the Visual System Allows the Adaptation of Evening Activity to Long Photoperiods in Drosophila melanogaster.

In many animals, the circadian clock plays a role in adapting to the coming season by measuring day length. The mechanism for measuring day length and its neuronal circuits remains elusive, however. Under laboratory conditions, the fruit fly, Drosophila melanogaster, displays 2 activity peaks: one in the morning and one in the evening. These peaks appear to be regulated by 2 separate circadian oscillators (the morning and evening oscillators) that reside in different subsets of pacemaker clock neurons in the brain. The morning and evening activity peaks can flexibly change their phases to adapt to different photoperiods by tracking dawn and dusk, respectively. In this study, we found that cryptochrome (CRY) in the evening oscillators (the fifth small ventral lateral neuron [5th s-LNv] and the dorsal lateral neurons [LNds]) limits the ability of the evening peak to track dusk during long days. In contrast, light signaling from the external photoreceptors (compound eyes, ocelli, and Hofbauer-Buchner eyelets) increases the ability of the evening peak to track dusk. At the molecular level, CRY signaling dampens the amplitude of PAR-domain protein 1 (PDP1) oscillations in most clock neurons during long days, whereas signaling from the visual system increases these amplitudes. Thus, our results suggest that light inputs from the two major circadian photoreceptors, CRY and the visual system, have opposite effects on day length adaptation. Their tug-of-war appears to determine the precise phase adjustment of evening activity.

A New Rhodopsin Influences Light-dependent Daily Activity Patterns of Fruit Flies.

Rhodopsin 7 ( Rh7), a new invertebrate Rhodopsin gene, was discovered in the genome of Drosophila melanogaster in 2000, but its function has remained elusive. We generated an Rh7 null mutant ( Rh70) by P element-mediated mutagenesis and found that an absence of Rh7 had significant effects on fly activity patterns during light-dark (LD) cycles: Rh70 mutants exhibited less morning activity and a longer siesta than wild-type controls. Consistent with these results, we found that Rh7 appears to be expressed in a few dorsal clock neurons that have been previously implicated in the control of the siesta. We also found putative Rh7 expression in R8 photoreceptor cells of the compound eyes and in the Hofbauer-Buchner eyelets, which have been shown to control the precise timing of locomotor activity. The absence of Rh7 alone impaired neither the flies' responses to constant white light nor the ability to follow phase shifts of white LD cycles. However, in blue light (470 nm), Rh70 mutants needed significantly longer to synchronize than wild-type controls, suggesting that Rh7 is a blue light-sensitive photopigment with a minor contribution to circadian clock synchronization. In combination with mutants that lacked additionally cryptochrome-based and/or eye-based light input to the circadian clock, the absence of Rh7 provoked slightly stronger effects.

Drosophila Rhodopsin 7 can partially replace the structural role of Rhodopsin 1, but not its physiological function.

Rhodopsin 7 (Rh7), a new invertebrate Rhodopsin gene, was discovered in the genome of Drosophila melanogaster in 2000 and thought to encode for a functional Rhodopsin protein. Indeed, Rh7 exhibits most hallmarks of the known Rhodopsins, except for the G-protein-activating QAKK motif in the third cytoplasmic loop that is absent in Rh7. Here, we show that Rh7 can partially substitute Rh1 in the outer receptor cells (R1-6) for rhabdomere maintenance, but that it cannot activate the phototransduction cascade in these cells. This speaks against a role of Rh7 as photopigment in R1-6, but does not exclude that it works in the inner photoreceptor cells.

Cryptochrome-dependent and -independent circadian entrainment circuits in Drosophila.

Entrainment to environmental light/dark (LD) cycles is a central function of circadian clocks. In Drosophila, entrainment is achieved by Cryptochrome (CRY) and input from the visual system. During activation by brief light pulses, CRY triggers the degradation of TIMELESS and subsequent shift in circadian phase. This is less important for LD entrainment, leading to questions regarding light input circuits and mechanisms from the visual system. Recent studies show that different subsets of brain pacemaker clock neurons, the morning (M) and evening (E) oscillators, have distinct functions in light entrainment. However, the role of CRY in M and E oscillators for entrainment to LD cycles is unknown. Here, we address this question by selectively expressing CRY in different subsets of clock neurons in a cry-null (cry(0)) mutant background. We were able to rescue the light entrainment deficits of cry(0) mutants by expressing CRY in E oscillators but not in any other clock neurons. Par domain protein 1 molecular oscillations in the E, but not M, cells of cry(0) mutants still responded to the LD phase delay. This residual light response was stemming from the visual system because it disappeared when all external photoreceptors were ablated genetically. We concluded that the E oscillators are the targets of light input via CRY and the visual system and are required for normal light entrainment.

Phase-shifting the fruit fly clock without cryptochrome.

The blue light photopigment cryptochrome (CRY) is thought to be the main circadian photoreceptor of Drosophila melanogaster. Nevertheless, entrainment to light-dark cycles is possible without functional CRY. Here, we monitored phase response curves of cry(01) mutants and control flies to 1-hour 1000-lux light pulses. We found that cry(01) mutants phase-shift their activity rhythm in the subjective early morning and late evening, although with reduced magnitude. This phase-shifting capability is sufficient for the slowed entrainment of the mutants, indicating that the eyes contribute to the clock's light sensitivity around dawn and dusk. With longer light pulses (3 hours and 6 hours), wild-type flies show greatly enhanced magnitude of phase shift, but CRY-less flies seem impaired in the ability to integrate duration of the light pulse in a wild-type manner: Only 6-hour light pulses at circadian time 21 significantly increased the magnitude of phase advances in cry(01) mutants. At circadian time 15, the mutants exhibited phase advances instead of the expected delays. These complex results are discussed.