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      Contribution of non-circadian neurons to the temporal organization of locomotor activity

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          ABSTRACT

          In the fruit fly, Drosophila melanogaster, the daily cycle of rest and activity is a rhythmic behavior that relies on the activity of a small number of neurons. The small ventral lateral neurons (sLNvs) are considered key in the control of locomotor rhythmicity. Previous work from our laboratory has showed that these neurons undergo structural remodeling on their axonal projections on a daily basis. Such remodeling endows sLNvs with the possibility to make synaptic contacts with different partners at different times throughout the day, as has been previously described. By using different genetic tools to alter membrane excitability of the sLNv putative postsynaptic partners, we tested their functional role in the control of locomotor activity. We also used optical imaging to test the functionality of these contacts. We found that these different neuronal groups affect the consolidation of rhythmic activity, suggesting that non-circadian cells are part of the circuit that controls locomotor activity. Our results suggest that new neuronal groups, in addition to the well-characterized clock neurons, contribute to the operations of the circadian network that controls locomotor activity in D. melanogaster.

          Abstract

          Summary: Here we characterized the impact of different putative postsynaptic partners of the sLNvs on the control of rhythmic locomotor behavior. We found that some of these novel neuronal clusters are relevant for the control of locomotor activity.

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          Most cited references54

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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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            GFP Reconstitution Across Synaptic Partners (GRASP) defines cell contacts and synapses in living nervous systems.

            The identification of synaptic partners is challenging in dense nerve bundles, where many processes occupy regions beneath the resolution of conventional light microscopy. To address this difficulty, we have developed GRASP, a system to label membrane contacts and synapses between two cells in living animals. Two complementary fragments of GFP are expressed on different cells, tethered to extracellular domains of transmembrane carrier proteins. When the complementary GFP fragments are fused to ubiquitous transmembrane proteins, GFP fluorescence appears uniformly along membrane contacts between the two cells. When one or both GFP fragments are fused to synaptic transmembrane proteins, GFP fluorescence is tightly localized to synapses. GRASP marks known synaptic contacts in C. elegans, correctly identifies changes in mutants with altered synaptic specificity, and can uncover new information about synaptic locations as confirmed by electron microscopy. GRASP may prove particularly useful for defining connectivity in complex nervous systems.
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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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                Author and article information

                Journal
                Biol Open
                Biol Open
                BIO
                biolopen
                Biology Open
                The Company of Biologists Ltd
                2046-6390
                15 January 2019
                10 December 2018
                10 December 2018
                : 8
                : 1
                : bio039628
                Affiliations
                Laboratorio de Genética del Comportamiento, Fundación Instituto Leloir and Instituto de Investigaciones Bioquímicas–Buenos Aires (IIB–BA, CONICET) , 1425 Buenos Aires, Argentina
                Author notes
                [*]

                Present address: Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Fisiología, Biología Molecular y Celular and CONICET–Universidad de Buenos Aires, Instituto de Fisiología, Biología Molecular y Neurociencias (IFIByNE), 1428 Buenos Aires, Argentina.

                [‡]

                Present address: Unidad de Transferencia Genética. Instituto de Oncología Ángel H. Roffo. 1417 Buenos Aires, Argentina.

                [§ ]Author for correspondence ( npirez@ 123456fbmc.fcen.uba.ar )
                Author information
                http://orcid.org/0000-0002-0463-0080
                http://orcid.org/0000-0002-5186-2407
                http://orcid.org/0000-0001-8945-3070
                Article
                BIO039628
                10.1242/bio.039628
                6361196
                30530810
                da367623-32f5-4aaa-9af6-d4fc925b0d08
                © 2019. Published by The Company of Biologists Ltd

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

                History
                : 23 October 2018
                : 3 December 2018
                Funding
                Funded by: Fondo para la Investigación Científica y Tecnológica, http://dx.doi.org/10.13039/501100006668;
                Award ID: PICT2011-2185
                Award ID: PICT2012-1230
                Categories
                Research Article

                Life sciences
                drosophila,slnvs,connectivity,non-circadian neurons,locomotor rhythms
                Life sciences
                drosophila, slnvs, connectivity, non-circadian neurons, locomotor rhythms

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