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      A tonically active master neuron modulates mutually exclusive motor states at two timescales

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          Abstract

          Continuity of behaviors requires animals to make smooth transitions between mutually exclusive behavioral states. Neural principles that govern these transitions are not well understood. Caenorhabditis elegans spontaneously switch between two opposite motor states, forward and backward movement, a phenomenon thought to reflect the reciprocal inhibition between interneurons AVB and AVA. Here, we report that spontaneous locomotion and their corresponding motor circuits are not separately controlled. AVA and AVB are neither functionally equivalent nor strictly reciprocally inhibitory. AVA, but not AVB, maintains a depolarized membrane potential. While AVA phasically inhibits the forward promoting interneuron AVB at a fast timescale, it maintains a tonic, extrasynaptic excitation on AVB over the longer timescale. We propose that AVA, with tonic and phasic activity of opposite polarities on different timescales, acts as a master neuron to break the symmetry between the underlying forward and backward motor circuits. This master neuron model offers a parsimonious solution for sustained locomotion consisted of mutually exclusive motor states.

          Abstract

          Mutually exclusive motor states are governed by a single neuron acting at two timescales in C. elegans.

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          Most cited references105

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          Genome engineering using the CRISPR-Cas9 system.

          Targeted nucleases are powerful tools for mediating genome alteration with high precision. The RNA-guided Cas9 nuclease from the microbial clustered regularly interspaced short palindromic repeats (CRISPR) adaptive immune system can be used to facilitate efficient genome engineering in eukaryotic cells by simply specifying a 20-nt targeting sequence within its guide RNA. Here we describe a set of tools for Cas9-mediated genome editing via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, as well as generation of modified cell lines for downstream functional studies. To minimize off-target cleavage, we further describe a double-nicking strategy using the Cas9 nickase mutant with paired guide RNAs. This protocol provides experimentally derived guidelines for the selection of target sites, evaluation of cleavage efficiency and analysis of off-target activity. Beginning with target design, gene modifications can be achieved within as little as 1-2 weeks, and modified clonal cell lines can be derived within 2-3 weeks.
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            Genome sequence of the nematode C. elegans: a platform for investigating biology.

            (1999)
            The 97-megabase genomic sequence of the nematode Caenorhabditis elegans reveals over 19,000 genes. More than 40 percent of the predicted protein products find significant matches in other organisms. There is a variety of repeated sequences, both local and dispersed. The distinctive distribution of some repeats and highly conserved genes provides evidence for a regional organization of the chromosomes.
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              The Structure of the Nervous System of the Nematode Caenorhabditis elegans

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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: ValidationRole: VisualizationRole: Writing - original draftRole: Writing - review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SoftwareRole: SupervisionRole: ValidationRole: VisualizationRole: Writing - original draftRole: Writing - review & editing
                Role: InvestigationRole: Validation
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: Validation
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: ValidationRole: VisualizationRole: Writing - original draftRole: Writing - review & editing
                Role: Resources
                Role: Resources
                Role: ConceptualizationRole: Funding acquisitionRole: ResourcesRole: Validation
                Role: ConceptualizationRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: Writing - review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: ValidationRole: VisualizationRole: Writing - review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing - original draftRole: Writing - review & editing
                Journal
                Sci Adv
                Sci Adv
                sciadv
                advances
                Science Advances
                American Association for the Advancement of Science
                2375-2548
                12 April 2024
                10 April 2024
                : 10
                : 15
                : eadk0002
                Affiliations
                [ 1 ]Department of Physiology, University of Toronto, Toronto, ON, Canada.
                [ 2 ]Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada.
                [ 3 ]College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.
                [ 4 ]University Claude Bernard Lyon 1, MeLiS, CNRS UMR 5284, INSERM U1314, 69008 Lyon, France.
                [ 5 ]Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
                Author notes
                [* ]Corresponding author. Email: ahamedt@ 123456hhmi.org (T.A.); sgao@ 123456hust.edu.cn (S.G.); meizhen@ 123456lunenfeld.ca (M.Z.)
                [†]

                Present address: Department of Biology, KU Leuven, Leuven, Belgium.

                [‡]

                Present address: HHMI Janelia Research Campus, Ashburn, VA, USA.

                [§]

                Present address: Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, China.

                [¶]

                Present address: King Abdullah University of Science and Technology (KAUST), Biological and Environmental Science and Engineering Division (BESE), KAUST Environmental Epigenetics Program (KEEP), Thuwal 23955-6900, Saudi Arabia.

                Author information
                https://orcid.org/0000-0002-4165-8496
                https://orcid.org/0000-0002-1405-3539
                https://orcid.org/0000-0002-2907-2169
                https://orcid.org/0000-0001-8488-7638
                https://orcid.org/0009-0006-1401-9871
                https://orcid.org/0000-0003-0268-8403
                https://orcid.org/0000-0002-1734-1915
                https://orcid.org/0000-0001-5431-4628
                https://orcid.org/0000-0003-0086-9622
                Article
                adk0002
                10.1126/sciadv.adk0002
                11006214
                38598630
                e06f06bd-9aaa-47ee-9a80-f4de5687ca4a
                Copyright © 2024 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

                History
                : 27 July 2023
                : 07 March 2024
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100010663, H2020 European Research Council;
                Award ID: 337702-K
                Funded by: FundRef http://dx.doi.org/10.13039/501100000038, Natural Sciences and Engineering Research Council of Canada;
                Award ID: RGPIN2017-06738
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 31871069
                Funded by: Canadian institute of Health Research;
                Award ID: FDN154274
                Funded by: the key International (Regional) Joint Research Project;
                Award ID: 32020103007
                Funded by: the key International (Regional) Joint Research Project;
                Award ID: 32020103007
                Categories
                Research Article
                Neuroscience
                SciAdv r-articles
                Biophysics
                Neuroscience
                Neuroscience
                Custom metadata
                Lou Notario

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