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      A rhodopsin in the brain functions in circadian photoentrainment in Drosophila

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      1 , 2 , 3 , 3 , 1
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          Abstract

          Animals partition their daily activity rhythms through their internal circadian clocks, which are synchronized by oscillating day-night cycles of light. The fruit fly, Drosophila melanogaster, senses day/night cycles in part through rhodopsin-dependent light reception in the compound eye, and photoreceptor cells in the Hofbauer-Buchner (H-B) eyelet 1 . However, a more significant light entrainment pathway is mediated in central pacemaker neurons in the brain. The Drosophila circadian clock is extremely light sensitive. However, the only known light sensor in pacemaker neurons, the flavoprotein, cryptochrome (Cry) 2, 3 , responds only to high levels of light in vitro 4 . These observations indicate the existence of an additional light-sensing pathway in fly pacemaker neurons 5 . Here, we identified an uncharacterized rhodopsin, Rh7, which functions in circadian light entrainment through circadian pacemaker neurons in the brain. The pacemaker neurons respond to violet light, which was dependent on Rh7. While loss of either cry or rh7 caused minor affects on photoentrainment, the defects in the double mutant were profound. The circadian photoresponse to constant light was impaired in the rh7 mutant, especially under dim light. The demonstration that Rh7 functions in circadian pacemaker neurons represents the first role for an opsin in the central brain.

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          Most cited references39

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          A pdf neuropeptide gene mutation and ablation of PDF neurons each cause severe abnormalities of behavioral circadian rhythms in Drosophila.

          The mechanisms by which circadian pacemaker systems transmit timing information to control behavior are largely unknown. Here, we define two critical features of that mechanism in Drosophila. We first describe animals mutant for the pdf neuropeptide gene, which is expressed by most of the candidate pacemakers (LNv neurons). Next, we describe animals in which pdf neurons were selectively ablated. Both sets of animals produced similar behavioral phenotypes. Both sets entrained to light, but both were largely arrhythmic under constant conditions. A minority of each pdf variant exhibited weak to moderate free-running rhythmicity. These results confirm the assignment of LNv neurons as the principal circadian pacemakers controlling daily locomotion in Drosophila. They also implicate PDF as the principal circadian transmitter.
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            CRY, a Drosophila clock and light-regulated cryptochrome, is a major contributor to circadian rhythm resetting and photosensitivity.

            Light is a major environmental signal for circadian rhythms. We have identified and analyzed cry, a novel Drosophila cryptochrome gene. All characterized family members are directly photosensitive and include plant blue light photoreceptors. We show that cry transcription is under circadian regulation, influenced by the Drosophila clock genes period, timeless, Clock, and cycle. We also show that cry protein levels are dramatically affected by light exposure. Importantly, circadian photosensitivity is increased in a cry-overexpressing strain. These physiological and genetic data therefore link a specific photoreceptor molecule to circadian rhythmicity. Taken together with the data in the accompanying paper, we propose that CRY is a major Drosophila photoreceptor dedicated to the resetting of circadian rhythms.
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              Versatile P(acman) BAC Libraries for Transgenesis Studies in Drosophila melanogaster

              We constructed Drosophila melanogaster BAC libraries with 21-kb and 83-kb inserts in the P(acman) system. Clones representing 12-fold coverage and encompassing more than 95% of annotated genes were mapped onto the reference genome. These clones can be integrated into predetermined attP sites in the genome using ΦC31 integrase to rescue mutations. They can be modified through recombineering, for example to incorporate protein tags and assess expression patterns.
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                Author and article information

                Journal
                0410462
                6011
                Nature
                Nature
                Nature
                0028-0836
                1476-4687
                14 April 2017
                10 May 2017
                18 May 2017
                10 November 2017
                : 545
                : 7654
                : 340-344
                Affiliations
                [1 ]Neuroscience Research Institute and Department of Molecular, Cellular and Developmental Biology, University of California Santa Barbara, Santa Barbara, CA, 93106, USA
                [2 ]Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
                [3 ]Department of Physiology and Biophysics, University of California, Irvine, Irvine, California, USA
                Author notes
                [4 ]Correspondence: cmontell@ 123456lifesci.ucsb.edu , phone, (805) 893-3634
                Article
                NIHMS864507
                10.1038/nature22325
                5476302
                28489826
                ce9d825d-ec59-493f-814e-73545c24abe6

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