[{"title":"Lineage origin of spinal cord cell type diversity","acknowledgement":"We would like to thank Elizabeth Marin, Anna Kicheva, Igor Adameyko, and James Briscoe as\r\nwell as members of the Sweeney and Hippemeyer labs and SFB consortium for comments on\r\nthe manuscript. We are also grateful for the technical support of the Preclinical and Imaging and\r\nOptics Facilities support teams (ISTA). In addition, we thank our funding sources for providing\r\nthe resources to do these experiments: Horizon Europe ERC Starting Grant Number 101041551\r\n(M.S.; L.B.S.); Special Research Program (SFB) of the Austrian Science Fund (FWF)\r\nNeuroStem Modulation Project numbers F7814-B (S.A.G.; M.S.; G.S.; and L.B.S.) and F7805\r\n(G.C. and S.H.). S.A.G is supported by a Boehringer Ingelheim Fonds PhD Fellowship, F.D.S.N.\r\nby an Institute of Science and Technology Austria (ISTA) GROW fellowship, and G.C. by an\r\nISTA Plus postdoctoral fellowship from the European Commission. S.H./L.B.S. and G.C. were\r\nadditionally supported by institutional funds from the ISTA and the University of Exeter,\r\nrespectively. ","ddc":["570"],"date_updated":"2026-04-14T08:16:55Z","_id":"21291","department":[{"_id":"SiHi"},{"_id":"LoSw"}],"has_accepted_license":"1","corr_author":"1","abstract":[{"lang":"eng","text":"The complexity and specificity of movement in vertebrates is driven by a rich diversity of spinal motor and interneuron cell types. During development, eleven spinal cord progenitor domains generate an equivalent number of cardinal neuron types. How progenitor domains, individual progenitors, and post-mitotic diversity relate is still unknown. We performed high-resolution, single-progenitor cell lineage tracing in the embryonic mouse spinal cord using mosaic analysis with double markers (MADM). Our quantitative study of lineage progression revealed that spinal cord progenitors undergo highly variable numbers of proliferative, neurogenic, and gliogenic cell divisions. The nascent clonally-related neurons migrate radially over large distances, span the dorsoventral axis, and even cross the midline, demonstrating striking bilaterality. Molecular and morphometric analysis indicate high levels of progenitor multipotency, with an individual progenitor capable of producing several molecularly and morphologically distinct neuron types, as well as astrocytes. These findings redefine spinal cord development as a process in which lineage variability — rather than rigid progenitor identity — drives the generation of cellular diversity."}],"doi":"10.64898/2026.02.12.705305","citation":{"mla":"Gobeil, Sophie A., et al. “Lineage Origin of Spinal Cord Cell Type Diversity.” <i>BioRxiv</i>, doi:<a href=\"https://doi.org/10.64898/2026.02.12.705305\">10.64898/2026.02.12.705305</a>.","apa":"Gobeil, S. A., Da Silveira Neto, F., Silvestrelli, G., Smits, M. G., Streicher, C., Cheung, G. T., … Sweeney, L. B. (n.d.). Lineage origin of spinal cord cell type diversity. <i>bioRxiv</i>. <a href=\"https://doi.org/10.64898/2026.02.12.705305\">https://doi.org/10.64898/2026.02.12.705305</a>","ieee":"S. A. Gobeil <i>et al.</i>, “Lineage origin of spinal cord cell type diversity,” <i>bioRxiv</i>. .","ista":"Gobeil SA, Da Silveira Neto F, Silvestrelli G, Smits MG, Streicher C, Cheung GT, Hippenmeyer S, Sweeney LB. Lineage origin of spinal cord cell type diversity. bioRxiv, <a href=\"https://doi.org/10.64898/2026.02.12.705305\">10.64898/2026.02.12.705305</a>.","chicago":"Gobeil, Sophie A, Francisco Da Silveira Neto, Giulia Silvestrelli, Matthijs Geert Smits, Carmen Streicher, Giselle T Cheung, Simon Hippenmeyer, and Lora B. Sweeney. “Lineage Origin of Spinal Cord Cell Type Diversity.” <i>BioRxiv</i>, n.d. <a href=\"https://doi.org/10.64898/2026.02.12.705305\">https://doi.org/10.64898/2026.02.12.705305</a>.","short":"S.A. Gobeil, F. Da Silveira Neto, G. Silvestrelli, M.G. Smits, C. Streicher, G.T. Cheung, S. Hippenmeyer, L.B. Sweeney, BioRxiv (n.d.).","ama":"Gobeil SA, Da Silveira Neto F, Silvestrelli G, et al. Lineage origin of spinal cord cell type diversity. <i>bioRxiv</i>. doi:<a href=\"https://doi.org/10.64898/2026.02.12.705305\">10.64898/2026.02.12.705305</a>"},"article_processing_charge":"No","project":[{"grant_number":"101041551","name":"Development and Evolution of Tetrapod Motor Circuits","_id":"ebb66355-77a9-11ec-83b8-b8ac210a4dae"},{"grant_number":"F7814","name":"Stem Cell Modulation in Neural Development and Regeneration/ P14-Swim-to-limb transition: cell type to connection diversity","_id":"8da85f50-16d5-11f0-9cad-eab8b0ff6c9e"},{"grant_number":"F7805","_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression"}],"language":[{"iso":"eng"}],"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","month":"02","oa_version":"Preprint","main_file_link":[{"url":"https://doi.org/10.64898/2026.02.12.705305","open_access":"1"}],"date_published":"2026-02-16T00:00:00Z","oa":1,"date_created":"2026-02-17T11:36:20Z","type":"preprint","status":"public","year":"2026","tmp":{"image":"/images/cc_by_nc_nd.png","short":"CC BY-NC-ND (4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)"},"publication":"bioRxiv","fulldoi":"https://doi.org/10.64898/2026.02.12.705305","publication_status":"submitted","OA_type":"green","day":"16","acknowledged_ssus":[{"_id":"PreCl"},{"_id":"Bio"}],"author":[{"first_name":"Sophie A","full_name":"Gobeil, Sophie A","last_name":"Gobeil","id":"2f3e9efb-eb24-11ec-86b2-88efb11d59fa"},{"last_name":"Da Silveira Neto","id":"8cfb7412-10a7-11f1-add1-82b44e6418f2","first_name":"Francisco","full_name":"Da Silveira Neto, Francisco"},{"last_name":"Silvestrelli","id":"12632ae8-799e-11ef-94a2-e5a3b5ef49e9","first_name":"Giulia","full_name":"Silvestrelli, Giulia"},{"first_name":"Matthijs Geert","full_name":"Smits, Matthijs Geert","id":"7a231d52-e216-11ee-a0bb-8acd55f8f1f0","last_name":"Smits"},{"last_name":"Streicher","id":"36BCB99C-F248-11E8-B48F-1D18A9856A87","first_name":"Carmen","full_name":"Streicher, Carmen"},{"last_name":"Cheung","orcid":"0000-0001-8457-2572","id":"471195F6-F248-11E8-B48F-1D18A9856A87","full_name":"Cheung, Giselle T","first_name":"Giselle T"},{"id":"37B36620-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0003-2279-1061","last_name":"Hippenmeyer","full_name":"Hippenmeyer, Simon","first_name":"Simon"},{"id":"56BE8254-C4F0-11E9-8E45-0B23E6697425","orcid":"0000-0001-9242-5601","last_name":"Sweeney","first_name":"Lora Beatrice Jaeger","full_name":"Sweeney, Lora Beatrice Jaeger"}],"OA_place":"repository"},{"title":"Temporal uncoupling of radial glia lineage progression in cortical organoids","ddc":["570"],"date_updated":"2026-09-09T07:11:02Z","publisher":"Springer Nature","_id":"22753","external_id":{"pmid":["42587153"]},"acknowledgement":"We thank M. L. de Guevara, S. Jayaram and A. Heger for technical assistance with mESC derivation; M. Goudarzi for assistance with organoid imaging; M. Leeb and F. Freeman for advice in culturing mESCs and organoids; S. Gobeil and L. Sweeney for reagents and advice for organoid clearing; A. Heger for mouse colony management; J. Hauser for technical assistance; the Stanford Brain Organogenesis Workshop; and all members of the Hippenmeyer laboratory for discussion and/or comments on the manuscript. This study was supported by the Scientific Service Units (SSU) of the Institute of Science and Technology, Austria through resources provided by the Imaging and Optics Facility (IOF), Laboratory Support Facility (LSF) and Preclinical Facility (PCF). M.S. received funding from the European Commission (IST plus postdoctoral fellowship). This work was supported by ISTA institutional funds to S.H., FWF SFB F78 Neuro Stem Modulation to S.H., and by the European Research Council (ERC) under the European Union’s Horizon 2020 Research And Innovation Program (grant agreement 725780 LinPro) to S.H. Open access funding provided by Institute of Science and Technology (IST Austria).","corr_author":"1","department":[{"_id":"SiHi"}],"has_accepted_license":"1","article_processing_charge":"Yes (via OA deal)","abstract":[{"lang":"eng","text":"Radial glial progenitors (RGPs) produce all excitatory neurons in the developing cerebral cortex. Mosaic analysis with double markers (MADM)-based lineage tracing in vivo has revealed a quantitative framework of RGP lineage progression1. Here we established MADM technology2,3 in mouse embryonic stem cells to probe RGP lineage progression in a self-organizing cortical organoid system. We found that RGPs exhibit a high level of plasticity in proliferative potential in organoids rather than strict temporally stereotyped lineage progression as observed in vivo. RGPs in organoids showed increased lineage restriction, diminishing cell-type diversity in clones of cortical projection neurons, despite uniform single-cell transcriptional signatures of RGPs and a unitary lineage trajectory. Thus, critical non-cell-autonomous cues that are absent in self-organizing systems and/or the genuine stem cell niche are essential for faithful temporal control of RGP lineage progression and the generation of clonal cortical cell-type diversity."}],"publication_identifier":{"issn":["0028-0836"],"eissn":["1476-4687"]},"doi":"10.1038/s41586-026-10916-7","file_date_updated":"2026-09-07T13:24:10Z","citation":{"chicago":"Stouffer, Melissa A, Osvaldo Miranda, Florian Pauler, Fabrizia Pipicelli, Carmen Streicher, Giselle T Cheung, and Simon Hippenmeyer. “Temporal Uncoupling of Radial Glia Lineage Progression in Cortical Organoids.” <i>Nature</i>. Springer Nature, 2026. <a href=\"https://doi.org/10.1038/s41586-026-10916-7\">https://doi.org/10.1038/s41586-026-10916-7</a>.","short":"M.A. Stouffer, O. Miranda, F. Pauler, F. Pipicelli, C. Streicher, G.T. Cheung, S. Hippenmeyer, Nature (2026).","ama":"Stouffer MA, Miranda O, Pauler F, et al. Temporal uncoupling of radial glia lineage progression in cortical organoids. <i>Nature</i>. 2026. doi:<a href=\"https://doi.org/10.1038/s41586-026-10916-7\">10.1038/s41586-026-10916-7</a>","ieee":"M. A. Stouffer <i>et al.</i>, “Temporal uncoupling of radial glia lineage progression in cortical organoids,” <i>Nature</i>. Springer Nature, 2026.","ista":"Stouffer MA, Miranda O, Pauler F, Pipicelli F, Streicher C, Cheung GT, Hippenmeyer S. 2026. Temporal uncoupling of radial glia lineage progression in cortical organoids. Nature.","mla":"Stouffer, Melissa A., et al. “Temporal Uncoupling of Radial Glia Lineage Progression in Cortical Organoids.” <i>Nature</i>, Springer Nature, 2026, doi:<a href=\"https://doi.org/10.1038/s41586-026-10916-7\">10.1038/s41586-026-10916-7</a>.","apa":"Stouffer, M. A., Miranda, O., Pauler, F., Pipicelli, F., Streicher, C., Cheung, G. T., &#38; Hippenmeyer, S. (2026). Temporal uncoupling of radial glia lineage progression in cortical organoids. <i>Nature</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41586-026-10916-7\">https://doi.org/10.1038/s41586-026-10916-7</a>"},"dataavailabilitystatement":"All data generated and analysed in this study are included in the paper, source data and/or Supplementary Tables 2 and 3. Raw sequencing data have been deposited with Gene Expression Omnibus (GEO) accession number GSE327470. Source data are provided with this paper. All scripts used to prepare data and figures for this manuscript are accessible on GitHub at https://github.com/fpauler/Temporal-Uncoupling-of-Radial-Glia-Lineage-Progression-in-Cortical-Organoid.","project":[{"_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression","grant_number":"F7805"},{"_id":"260018B0-B435-11E9-9278-68D0E5697425","name":"Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development","call_identifier":"H2020","grant_number":"725780"}],"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","month":"08","language":[{"iso":"eng"}],"scopus_import":"1","oa_version":"Published Version","date_created":"2026-08-23T22:01:47Z","date_published":"2026-08-12T00:00:00Z","oa":1,"status":"public","PlanS_conform":"1","researchdata_availability":"yes","year":"2026","type":"journal_article","fulldoi":"https://doi.org/10.1038/s41586-026-10916-7","publication_status":"epub_ahead","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication":"Nature","supplementarymaterial":"yes","pmid":1,"OA_type":"hybrid","article_type":"original","file":[{"success":1,"file_id":"22844","content_type":"application/pdf","creator":"dernst","file_size":48513068,"file_name":"2026_Nature_Stouffer.pdf","access_level":"open_access","date_created":"2026-09-07T13:24:10Z","checksum":"11383e28fc430b28d2666f73833e8b56","relation":"main_file","date_updated":"2026-09-07T13:24:10Z"}],"quality_controlled":"1","author":[{"id":"4C9372C4-F248-11E8-B48F-1D18A9856A87","last_name":"Stouffer","full_name":"Stouffer, Melissa A","first_name":"Melissa A"},{"orcid":"0000-0001-6618-6889","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","last_name":"Miranda","full_name":"Miranda, Osvaldo","first_name":"Osvaldo"},{"last_name":"Pauler","orcid":"0000-0002-7462-0048","id":"48EA0138-F248-11E8-B48F-1D18A9856A87","full_name":"Pauler, Florian","first_name":"Florian"},{"last_name":"Pipicelli","id":"649134fd-d012-11ed-8f82-db1e5050f9ba","first_name":"Fabrizia","full_name":"Pipicelli, Fabrizia"},{"first_name":"Carmen","full_name":"Streicher, Carmen","id":"36BCB99C-F248-11E8-B48F-1D18A9856A87","last_name":"Streicher"},{"first_name":"Giselle T","full_name":"Cheung, Giselle T","orcid":"0000-0001-8457-2572","id":"471195F6-F248-11E8-B48F-1D18A9856A87","last_name":"Cheung"},{"first_name":"Simon","full_name":"Hippenmeyer, Simon","orcid":"0000-0003-2279-1061","last_name":"Hippenmeyer","id":"37B36620-F248-11E8-B48F-1D18A9856A87"}],"day":"12","acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"},{"_id":"PreCl"}],"OA_place":"publisher","ec_funded":1,"das_tickbox":"1"},{"quality_controlled":"1","OA_type":"closed access","pmid":1,"editor":[{"full_name":"Garcia-Marques, Jorge","first_name":"Jorge","last_name":"Garcia-Marques"},{"last_name":"Lee","first_name":"Tzumin","full_name":"Lee, Tzumin"}],"volume":2886,"page":"139-151","ec_funded":1,"acknowledged_ssus":[{"_id":"Bio"}],"day":"03","author":[{"id":"471195F6-F248-11E8-B48F-1D18A9856A87","last_name":"Cheung","orcid":"0000-0001-8457-2572","full_name":"Cheung, Giselle T","first_name":"Giselle T"},{"orcid":"0000-0002-7462-0048","id":"48EA0138-F248-11E8-B48F-1D18A9856A87","last_name":"Pauler","first_name":"Florian","full_name":"Pauler, Florian"},{"id":"37B36620-F248-11E8-B48F-1D18A9856A87","last_name":"Hippenmeyer","orcid":"0000-0003-2279-1061","first_name":"Simon","full_name":"Hippenmeyer, Simon"}],"type":"book_chapter","intvolume":"      2886","year":"2025","status":"public","date_published":"2025-01-03T00:00:00Z","date_created":"2025-01-07T08:36:47Z","publication":"Lineage Tracing","publication_status":"published","fulldoi":"https://doi.org/10.1007/978-1-0716-4310-5_7","project":[{"grant_number":"754411","_id":"260C2330-B435-11E9-9278-68D0E5697425","name":"ISTplus - Postdoctoral Fellowships","call_identifier":"H2020"}],"oa_version":"None","scopus_import":"1","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","month":"01","language":[{"iso":"eng"}],"acknowledgement":"We thank all Hippenmeyer lab members for support and discussions. Experimental steps described were optimized with support provided by the Imaging & Optics Facility (IOF) and Preclinical Facility (PCF) at ISTA, Vienna BioCenter Core Facilities (VBCF), and Christoph Bock lab at Center for Molecular Medicine (CeMM). G.C. received funding from European Commission (IST plus postdoctoral fellowship). This work was supported by ISTA institutional funds: The Austrian Science Fund Special Research Programmes (FWF SFB F78 Neuro Stem Modulation) to S.H.","place":"New York, NY","external_id":{"pmid":["39745639"]},"_id":"18765","publisher":"Springer Nature","date_updated":"2025-04-14T07:43:46Z","title":"Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM)","doi":"10.1007/978-1-0716-4310-5_7","citation":{"apa":"Cheung, G. T., Pauler, F., &#38; Hippenmeyer, S. (2025). Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM). In J. Garcia-Marques &#38; T. Lee (Eds.), <i>Lineage Tracing</i> (Vol. 2886, pp. 139–151). New York, NY: Springer Nature. <a href=\"https://doi.org/10.1007/978-1-0716-4310-5_7\">https://doi.org/10.1007/978-1-0716-4310-5_7</a>","mla":"Cheung, Giselle T., et al. “Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM).” <i>Lineage Tracing</i>, edited by Jorge Garcia-Marques and Tzumin Lee, vol. 2886, Springer Nature, 2025, pp. 139–51, doi:<a href=\"https://doi.org/10.1007/978-1-0716-4310-5_7\">10.1007/978-1-0716-4310-5_7</a>.","ama":"Cheung GT, Pauler F, Hippenmeyer S. Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM). In: Garcia-Marques J, Lee T, eds. <i>Lineage Tracing</i>. Vol 2886. MIMB. New York, NY: Springer Nature; 2025:139-151. doi:<a href=\"https://doi.org/10.1007/978-1-0716-4310-5_7\">10.1007/978-1-0716-4310-5_7</a>","short":"G.T. Cheung, F. Pauler, S. Hippenmeyer, in:, J. Garcia-Marques, T. Lee (Eds.), Lineage Tracing, Springer Nature, New York, NY, 2025, pp. 139–151.","chicago":"Cheung, Giselle T, Florian Pauler, and Simon Hippenmeyer. “Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM).” In <i>Lineage Tracing</i>, edited by Jorge Garcia-Marques and Tzumin Lee, 2886:139–51. MIMB. New York, NY: Springer Nature, 2025. <a href=\"https://doi.org/10.1007/978-1-0716-4310-5_7\">https://doi.org/10.1007/978-1-0716-4310-5_7</a>.","ista":"Cheung GT, Pauler F, Hippenmeyer S. 2025.Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM). In: Lineage Tracing. Methods in Molecular Biology, vol. 2886, 139–151.","ieee":"G. T. Cheung, F. Pauler, and S. Hippenmeyer, “Probing Cell-Type Specificity of Mutant Phenotype at Transcriptomic Level Using Mosaic Analysis with Double Markers (MADM),” in <i>Lineage Tracing</i>, vol. 2886, J. Garcia-Marques and T. Lee, Eds. New York, NY: Springer Nature, 2025, pp. 139–151."},"abstract":[{"lang":"eng","text":"Mosaic Analysis with Double Markers (MADM) represents a mouse genetic approach coupling differential fluorescent labeling to genetic manipulations in dividing cells and their lineages. MADM uniquely enables the generation and visualization of individual control or homozygous mutant cells in a heterozygous genetic environment. Among its diverse applications, MADM has been used to dissect cell-autonomous gene functions important for cortical development and neural development in general. The high cellular resolution offered by MADM also permits the analysis of transcriptomic changes of individual cells upon genetic manipulations. In this chapter, we describe an experimental protocol combining the generation and isolation of MADM-labeled cells with downstream single-cell RNA-sequencing technologies to probe cell-type specific phenotypes due to genetic mutations at single-cell resolution."}],"publication_identifier":{"eisbn":["9781071643105"],"issn":["1064-3745"],"eissn":["1940-6029"],"isbn":["9781071643099"]},"article_processing_charge":"No","department":[{"_id":"SiHi"}],"corr_author":"1","alternative_title":["Methods in Molecular Biology"],"series_title":"MIMB"},{"das_tickbox":"1","ec_funded":1,"day":"15","acknowledged_ssus":[{"_id":"Bio"},{"_id":"PreCl"}],"author":[{"full_name":"Amberg, Nicole","first_name":"Nicole","id":"4CD6AAC6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-3183-8207","last_name":"Amberg"},{"last_name":"Cheung","orcid":"0000-0001-8457-2572","id":"471195F6-F248-11E8-B48F-1D18A9856A87","full_name":"Cheung, Giselle T","first_name":"Giselle T"},{"orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","last_name":"Hippenmeyer","full_name":"Hippenmeyer, Simon","first_name":"Simon"}],"file":[{"relation":"main_file","date_updated":"2024-07-16T11:50:03Z","checksum":"3f0ee62e04bf5a44b45b035662826e95","date_created":"2024-07-16T11:50:03Z","access_level":"open_access","file_name":"2024_STARProtoc_Amberg.pdf","file_size":8871807,"creator":"dernst","file_id":"17260","content_type":"application/pdf","success":1}],"quality_controlled":"1","pmid":1,"article_type":"review","supplementarymaterial":"no","volume":5,"tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication":"STAR Protocols","fulldoi":"https://doi.org/10.1016/j.xpro.2023.102771","publication_status":"published","type":"journal_article","issue":"1","researchdata_availability":"not applicable","status":"public","year":"2024","intvolume":"         5","date_published":"2024-03-15T00:00:00Z","oa":1,"date_created":"2023-12-13T11:48:05Z","oa_version":"Published Version","scopus_import":"1","user_id":"317138e5-6ab7-11ef-aa6d-ffef3953e345","month":"03","language":[{"iso":"eng"}],"project":[{"grant_number":"T01031","call_identifier":"FWF","_id":"268F8446-B435-11E9-9278-68D0E5697425","name":"Role of Eed in neural stem cell lineage progression"},{"_id":"260C2330-B435-11E9-9278-68D0E5697425","name":"ISTplus - Postdoctoral Fellowships","call_identifier":"H2020","grant_number":"754411"},{"grant_number":"F7805","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression","_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E"},{"grant_number":"725780","name":"Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development","_id":"260018B0-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"}],"dataavailabilitystatement":"No custom code or dataset was generated in this protocol.","article_number":"102771","publication_identifier":{"issn":["2666-1667"]},"abstract":[{"text":"Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice and high-resolution phenotyping at the individual cell level. Here, we present a protocol for isolating MADM-labeled cells with high yield for downstream molecular analyses using fluorescence-activated cell sorting (FACS). We describe steps for generating MADM-labeled mice, perfusion, single-cell suspension, and debris removal. We then detail procedures for cell sorting by FACS and downstream analysis. This protocol is suitable for embryonic to adult mice.\r\nFor complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).1","lang":"eng"}],"citation":{"apa":"Amberg, N., Cheung, G. T., &#38; Hippenmeyer, S. (2024). Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. <i>STAR Protocols</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.xpro.2023.102771\">https://doi.org/10.1016/j.xpro.2023.102771</a>","mla":"Amberg, Nicole, et al. “Protocol for Sorting Cells from Mouse Brains Labeled with Mosaic Analysis with Double Markers by Flow Cytometry.” <i>STAR Protocols</i>, vol. 5, no. 1, 102771, Elsevier, 2024, doi:<a href=\"https://doi.org/10.1016/j.xpro.2023.102771\">10.1016/j.xpro.2023.102771</a>.","ista":"Amberg N, Cheung GT, Hippenmeyer S. 2024. Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. STAR Protocols. 5(1), 102771.","ieee":"N. Amberg, G. T. Cheung, and S. Hippenmeyer, “Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry,” <i>STAR Protocols</i>, vol. 5, no. 1. Elsevier, 2024.","ama":"Amberg N, Cheung GT, Hippenmeyer S. Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry. <i>STAR Protocols</i>. 2024;5(1). doi:<a href=\"https://doi.org/10.1016/j.xpro.2023.102771\">10.1016/j.xpro.2023.102771</a>","short":"N. Amberg, G.T. Cheung, S. Hippenmeyer, STAR Protocols 5 (2024).","chicago":"Amberg, Nicole, Giselle T Cheung, and Simon Hippenmeyer. “Protocol for Sorting Cells from Mouse Brains Labeled with Mosaic Analysis with Double Markers by Flow Cytometry.” <i>STAR Protocols</i>. Elsevier, 2024. <a href=\"https://doi.org/10.1016/j.xpro.2023.102771\">https://doi.org/10.1016/j.xpro.2023.102771</a>."},"file_date_updated":"2024-07-16T11:50:03Z","doi":"10.1016/j.xpro.2023.102771","article_processing_charge":"Yes (in subscription journal)","department":[{"_id":"SiHi"}],"has_accepted_license":"1","keyword":["General Immunology and Microbiology","General Biochemistry","Genetics and Molecular Biology","General Neuroscience"],"corr_author":"1","external_id":{"pmid":["38070137"]},"acknowledgement":"This research was supported by the Scientific Service Units (SSU) at IST Austria through resources provided by the Imaging & Optics Facility (IOF) and Preclinical Facilities (PCF). N.A. received support from FWF Firnberg-Programme (T 1031). G.C. received support from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 754411 as an ISTplus postdoctoral fellow. This work was also supported by IST Austria institutional funds, FWF SFB F78 to S.H., and the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 725780 LinPro) to S.H.","ddc":["570"],"date_updated":"2026-10-21T08:40:15Z","_id":"14683","publisher":"Elsevier","title":"Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry"},{"dataavailabilitystatement":"No dataset was generated while composing this protocol. A FIJI macro script “3DSerialSectionAlignmentLineROI.ijm” and a Microsoft Excel template used in this protocol are provided as supplementary materials.","article_number":"103157","project":[{"call_identifier":"H2020","name":"ISTplus - Postdoctoral Fellowships","_id":"260C2330-B435-11E9-9278-68D0E5697425","grant_number":"754411"},{"_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression","grant_number":"F7805"}],"month":"09","user_id":"317138e5-6ab7-11ef-aa6d-ffef3953e345","language":[{"iso":"eng"}],"scopus_import":"1","oa_version":"Published Version","title":"Protocol for quantitative reconstruction of cell lineage using mosaic analysis with double markers in mice","ddc":["570"],"date_updated":"2026-10-21T09:14:52Z","publisher":"Elsevier","_id":"17187","APC_amount":"804 EUR","external_id":{"pmid":["38935508"]},"acknowledgement":"We thank A. Heger for mouse breeding support. This work was supported by the Scientific Service Units of IST Austria through resources provided by the Imaging & Optics and Preclinical facilities. G.C. received funding from the European Commission (IST plus postdoctoral fellowship); S.H. was funded by ISTA institutional funds and the Austrian Science Fund Special Research Programmes (FWF SFB-F78 Neuro Stem Modulation).","corr_author":"1","department":[{"_id":"SiHi"}],"has_accepted_license":"1","article_processing_charge":"Yes","publication_identifier":{"eissn":["2666-1667"]},"abstract":[{"lang":"eng","text":"The generation of diverse cell types during development is fundamental to brain\r\nfunctions. We outline a protocol to quantitatively assess the clonal output of individual neural progenitors using mosaic analysis with double markers (MADM) in\r\nmice. We first describe steps to acquire and reconstruct adult MADM clones in\r\nthe superior colliculus. Then we detail analysis pipelines to determine clonal\r\ncomposition and architecture. This protocol enables the buildup of quantitative\r\nframeworks of lineage progression with precise spatial resolution in the brain.\r\nFor complete details on the use and execution of this protocol, please refer to\r\nCheung et al.1"}],"doi":"10.1016/j.xpro.2024.103157","citation":{"apa":"Cheung, G. T., Streicher, C., &#38; Hippenmeyer, S. (2024). Protocol for quantitative reconstruction of cell lineage using mosaic analysis with double markers in mice. <i>STAR Protocols</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.xpro.2024.103157\">https://doi.org/10.1016/j.xpro.2024.103157</a>","mla":"Cheung, Giselle T., et al. “Protocol for Quantitative Reconstruction of Cell Lineage Using Mosaic Analysis with Double Markers in Mice.” <i>STAR Protocols</i>, vol. 5, no. 3, 103157, Elsevier, 2024, doi:<a href=\"https://doi.org/10.1016/j.xpro.2024.103157\">10.1016/j.xpro.2024.103157</a>.","ama":"Cheung GT, Streicher C, Hippenmeyer S. Protocol for quantitative reconstruction of cell lineage using mosaic analysis with double markers in mice. <i>STAR Protocols</i>. 2024;5(3). doi:<a href=\"https://doi.org/10.1016/j.xpro.2024.103157\">10.1016/j.xpro.2024.103157</a>","short":"G.T. Cheung, C. Streicher, S. Hippenmeyer, STAR Protocols 5 (2024).","chicago":"Cheung, Giselle T, Carmen Streicher, and Simon Hippenmeyer. “Protocol for Quantitative Reconstruction of Cell Lineage Using Mosaic Analysis with Double Markers in Mice.” <i>STAR Protocols</i>. Elsevier, 2024. <a href=\"https://doi.org/10.1016/j.xpro.2024.103157\">https://doi.org/10.1016/j.xpro.2024.103157</a>.","ista":"Cheung GT, Streicher C, Hippenmeyer S. 2024. Protocol for quantitative reconstruction of cell lineage using mosaic analysis with double markers in mice. STAR Protocols. 5(3), 103157.","ieee":"G. T. Cheung, C. Streicher, and S. Hippenmeyer, “Protocol for quantitative reconstruction of cell lineage using mosaic analysis with double markers in mice,” <i>STAR Protocols</i>, vol. 5, no. 3. Elsevier, 2024."},"file_date_updated":"2025-01-09T12:12:40Z","supplementarymaterial":"yes","volume":5,"pmid":1,"article_type":"original","OA_type":"gold","file":[{"creator":"dernst","file_size":5186071,"success":1,"content_type":"application/pdf","file_id":"18809","checksum":"d8a8cdba82a394e731aa699ace1ae433","date_updated":"2025-01-09T12:12:40Z","relation":"main_file","file_name":"2024_STARProtoc_Cheung.pdf","access_level":"open_access","date_created":"2025-01-09T12:12:40Z"}],"quality_controlled":"1","author":[{"full_name":"Cheung, Giselle T","first_name":"Giselle T","id":"471195F6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8457-2572","last_name":"Cheung"},{"full_name":"Streicher, Carmen","first_name":"Carmen","last_name":"Streicher","id":"36BCB99C-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Simon","full_name":"Hippenmeyer, Simon","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","last_name":"Hippenmeyer"}],"day":"20","acknowledged_ssus":[{"_id":"Bio"},{"_id":"PreCl"}],"OA_place":"publisher","ec_funded":1,"das_tickbox":"1","date_created":"2024-06-30T22:01:04Z","date_published":"2024-09-20T00:00:00Z","oa":1,"year":"2024","status":"public","researchdata_availability":"yes (supplementary data)","intvolume":"         5","type":"journal_article","issue":"3","fulldoi":"https://doi.org/10.1016/j.xpro.2024.103157","publication_status":"published","tmp":{"image":"/images/cc_by_nc_nd.png","short":"CC BY-NC-ND (4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)"},"publication":"STAR Protocols"},{"intvolume":"         5","status":"public","year":"2024","researchdata_availability":"not applicable","issue":"3","type":"journal_article","date_created":"2024-07-14T22:01:10Z","oa":1,"date_published":"2024-09-20T00:00:00Z","publication_status":"published","fulldoi":"https://doi.org/10.1016/j.xpro.2024.103168","publication":"STAR Protocols","tmp":{"image":"/images/cc_by_nc_nd.png","short":"CC BY-NC-ND (4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode","name":"Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)"},"article_type":"original","OA_type":"gold","pmid":1,"quality_controlled":"1","file":[{"checksum":"464f52ecc6ec92f509552823bb82bf79","relation":"main_file","date_updated":"2025-01-09T12:16:53Z","file_name":"2024_STARProtoc_Cheung2.pdf","access_level":"open_access","date_created":"2025-01-09T12:16:53Z","creator":"dernst","file_size":6445556,"success":1,"file_id":"18810","content_type":"application/pdf"}],"volume":5,"supplementarymaterial":"no","OA_place":"publisher","das_tickbox":"1","author":[{"last_name":"Cheung","orcid":"0000-0001-8457-2572","id":"471195F6-F248-11E8-B48F-1D18A9856A87","first_name":"Giselle T","full_name":"Cheung, Giselle T"},{"orcid":"0000-0002-7462-0048","last_name":"Pauler","id":"48EA0138-F248-11E8-B48F-1D18A9856A87","first_name":"Florian","full_name":"Pauler, Florian"},{"orcid":"0000-0002-3509-1948","id":"3B8B25A8-F248-11E8-B48F-1D18A9856A87","last_name":"Koppensteiner","first_name":"Peter","full_name":"Koppensteiner, Peter"},{"id":"37B36620-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0003-2279-1061","last_name":"Hippenmeyer","full_name":"Hippenmeyer, Simon","first_name":"Simon"}],"acknowledged_ssus":[{"_id":"Bio"},{"_id":"M-Shop"},{"_id":"PreCl"}],"day":"20","_id":"17232","publisher":"Elsevier","date_updated":"2026-10-21T09:15:51Z","ddc":["570"],"acknowledgement":"We thank R. Beattie and T. Asenov for designing and producing components of the multi-well slice recover chamber. We thank R. Shigemoto for providing equipment access. We thank C. Streicher and A. Heger for mouse breeding support. This work was supported by the Scientific Service Units of IST Austria through resources provided by the Imaging & Optics, Miba Machine Shop, and Preclinical facilities. G.C. received funding from the European Commission (IST plus postdoctoral fellowship) and S.H. was funded by ISTA institutional funds and the Austrian Science Fund Special Research Programmes (FWF SFB-F78 Neuro Stem Modulation).","APC_amount":"804 EUR","external_id":{"pmid":["38968076"]},"title":"Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq","article_processing_charge":"Yes","file_date_updated":"2025-01-09T12:16:53Z","citation":{"ama":"Cheung GT, Pauler F, Koppensteiner P, Hippenmeyer S. Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq. <i>STAR Protocols</i>. 2024;5(3). doi:<a href=\"https://doi.org/10.1016/j.xpro.2024.103168\">10.1016/j.xpro.2024.103168</a>","short":"G.T. Cheung, F. Pauler, P. Koppensteiner, S. Hippenmeyer, STAR Protocols 5 (2024).","chicago":"Cheung, Giselle T, Florian Pauler, Peter Koppensteiner, and Simon Hippenmeyer. “Protocol for Mapping Cell Lineage and Cell-Type Identity of Clonally-Related Cells in Situ Using MADM-CloneSeq.” <i>STAR Protocols</i>. Elsevier, 2024. <a href=\"https://doi.org/10.1016/j.xpro.2024.103168\">https://doi.org/10.1016/j.xpro.2024.103168</a>.","ista":"Cheung GT, Pauler F, Koppensteiner P, Hippenmeyer S. 2024. Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq. STAR Protocols. 5(3), 103168.","ieee":"G. T. Cheung, F. Pauler, P. Koppensteiner, and S. Hippenmeyer, “Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq,” <i>STAR Protocols</i>, vol. 5, no. 3. Elsevier, 2024.","apa":"Cheung, G. T., Pauler, F., Koppensteiner, P., &#38; Hippenmeyer, S. (2024). Protocol for mapping cell lineage and cell-type identity of clonally-related cells in situ using MADM-CloneSeq. <i>STAR Protocols</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.xpro.2024.103168\">https://doi.org/10.1016/j.xpro.2024.103168</a>","mla":"Cheung, Giselle T., et al. “Protocol for Mapping Cell Lineage and Cell-Type Identity of Clonally-Related Cells in Situ Using MADM-CloneSeq.” <i>STAR Protocols</i>, vol. 5, no. 3, 103168, Elsevier, 2024, doi:<a href=\"https://doi.org/10.1016/j.xpro.2024.103168\">10.1016/j.xpro.2024.103168</a>."},"doi":"10.1016/j.xpro.2024.103168","publication_identifier":{"eissn":["2666-1667"]},"abstract":[{"lang":"eng","text":"The lineage relationship of clonally-related cells offers important insights into the ontogeny and cytoarchitecture of the brain in health and disease. Here, we provide a protocol to concurrently assess cell lineage relationship and cell-type identity among clonally-related cells in situ. We first describe the preparation and screening of acute brain slices containing clonally-related cells labeled using mosaic analysis with double markers (MADM). We then outline steps to collect RNA from individual cells for downstream applications and cell-type identification using RNA sequencing.\r\nFor complete details on the use and execution of this protocol, please refer to Cheung et al.\r\n1"}],"corr_author":"1","has_accepted_license":"1","department":[{"_id":"SiHi"},{"_id":"PreCl"}],"project":[{"grant_number":"F7805","_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression"}],"article_number":"103168","dataavailabilitystatement":"No dataset or new code was generated while composing this protocol.","oa_version":"Published Version","user_id":"317138e5-6ab7-11ef-aa6d-ffef3953e345","language":[{"iso":"eng"}],"month":"09","scopus_import":"1"},{"month":"01","language":[{"iso":"eng"}],"user_id":"317138e5-6ab7-11ef-aa6d-ffef3953e345","scopus_import":"1","oa_version":"Published Version","dataavailabilitystatement":"All data generated and analyzed in this study are included in the paper and/or supplementary materials. Raw sequencing data has been deposited at Gene Expression Omnibus (GEO). All scripts that were used to prepare data and figures for this manuscript are available via GitHub at https://github.com/fpauler. Accession numbers and github DOI are listed in the key resources table. Files for 3D printing of acute slice recovery chamber (3D model 361319) are deposited to https://www.printables.com.","project":[{"grant_number":"F7805","_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression"}],"corr_author":"1","has_accepted_license":"1","department":[{"_id":"SiHi"},{"_id":"RySh"}],"article_processing_charge":"Yes (via OA deal)","doi":"10.1016/j.neuron.2023.11.009","file_date_updated":"2024-02-06T13:56:15Z","citation":{"mla":"Cheung, Giselle T., et al. “Multipotent Progenitors Instruct Ontogeny of the Superior Colliculus.” <i>Neuron</i>, vol. 112, no. 2, Elsevier, 2024, p. 230–246.e11, doi:<a href=\"https://doi.org/10.1016/j.neuron.2023.11.009\">10.1016/j.neuron.2023.11.009</a>.","apa":"Cheung, G. T., Pauler, F., Koppensteiner, P., Krausgruber, T., Streicher, C., Schrammel, M., … Hippenmeyer, S. (2024). Multipotent progenitors instruct ontogeny of the superior colliculus. <i>Neuron</i>. Elsevier. <a href=\"https://doi.org/10.1016/j.neuron.2023.11.009\">https://doi.org/10.1016/j.neuron.2023.11.009</a>","short":"G.T. Cheung, F. Pauler, P. Koppensteiner, T. Krausgruber, C. Streicher, M. Schrammel, N.Y. Özgen, A. Ivec, C. Bock, R. Shigemoto, S. Hippenmeyer, Neuron 112 (2024) 230–246.e11.","chicago":"Cheung, Giselle T, Florian Pauler, Peter Koppensteiner, Thomas Krausgruber, Carmen Streicher, Martin Schrammel, Natalie Y Özgen, et al. “Multipotent Progenitors Instruct Ontogeny of the Superior Colliculus.” <i>Neuron</i>. Elsevier, 2024. <a href=\"https://doi.org/10.1016/j.neuron.2023.11.009\">https://doi.org/10.1016/j.neuron.2023.11.009</a>.","ama":"Cheung GT, Pauler F, Koppensteiner P, et al. Multipotent progenitors instruct ontogeny of the superior colliculus. <i>Neuron</i>. 2024;112(2):230-246.e11. doi:<a href=\"https://doi.org/10.1016/j.neuron.2023.11.009\">10.1016/j.neuron.2023.11.009</a>","ieee":"G. T. Cheung <i>et al.</i>, “Multipotent progenitors instruct ontogeny of the superior colliculus,” <i>Neuron</i>, vol. 112, no. 2. Elsevier, p. 230–246.e11, 2024.","ista":"Cheung GT, Pauler F, Koppensteiner P, Krausgruber T, Streicher C, Schrammel M, Özgen NY, Ivec A, Bock C, Shigemoto R, Hippenmeyer S. 2024. Multipotent progenitors instruct ontogeny of the superior colliculus. Neuron. 112(2), 230–246.e11."},"publication_identifier":{"issn":["0896-6273"]},"abstract":[{"text":"The superior colliculus (SC) in the mammalian midbrain is essential for multisensory integration and is composed of a rich diversity of excitatory and inhibitory neurons and glia. However, the developmental principles directing the generation of SC cell-type diversity are not understood. Here, we pursued systematic cell lineage tracing in silico and in vivo, preserving full spatial information, using genetic mosaic analysis with double markers (MADM)-based clonal analysis with single-cell sequencing (MADM-CloneSeq). The analysis of clonally related cell lineages revealed that radial glial progenitors (RGPs) in SC are exceptionally multipotent. Individual resident RGPs have the capacity to produce all excitatory and inhibitory SC neuron types, even at the stage of terminal division. While individual clonal units show no pre-defined cellular composition, the establishment of appropriate relative proportions of distinct neuronal types occurs in a PTEN-dependent manner. Collectively, our findings provide an inaugural framework at the single-RGP/-cell level of the mammalian SC ontogeny.","lang":"eng"}],"title":"Multipotent progenitors instruct ontogeny of the superior colliculus","_id":"12875","publisher":"Elsevier","date_updated":"2026-10-21T09:51:58Z","ddc":["570"],"acknowledgement":"We thank Liqun Luo for his continued support, for providing essential resources for generating Fzd10-CreER mice which were generated in his laboratory, and for comments on the manuscript; W. Zhong for providing Nestin-Cre transgenic mouse line for this study; A. Heger for mouse colony management; R. Beattie and T. Asenov for designing and producing components of acute slice recovery chamber for MADM-CloneSeq experiments; and K. Leopold, J. Rodarte and N. Amberg for initial experiments, technical support and/or assistance. This study was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Imaging & Optics Facility (IOF), Laboratory Support Facility (LSF), Miba Machine Shop, and Pre-clinical Facility (PCF). G.C. received funding from European Commission (IST plus postdoctoral fellowship). This work was supported by ISTA institutional\r\nfunds; the Austrian Science Fund Special Research Programmes (FWF SFB F78 Neuro Stem Modulation) to S.H. ","external_id":{"pmid":["38096816"],"isi":["001163937900001"]},"author":[{"last_name":"Cheung","id":"471195F6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8457-2572","first_name":"Giselle T","full_name":"Cheung, Giselle T"},{"id":"48EA0138-F248-11E8-B48F-1D18A9856A87","last_name":"Pauler","orcid":"0000-0002-7462-0048","first_name":"Florian","full_name":"Pauler, Florian"},{"id":"3B8B25A8-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-3509-1948","last_name":"Koppensteiner","full_name":"Koppensteiner, Peter","first_name":"Peter"},{"last_name":"Krausgruber","full_name":"Krausgruber, Thomas","first_name":"Thomas"},{"id":"36BCB99C-F248-11E8-B48F-1D18A9856A87","last_name":"Streicher","first_name":"Carmen","full_name":"Streicher, Carmen"},{"first_name":"Martin","full_name":"Schrammel, Martin","last_name":"Schrammel","id":"f13e7cae-e8bd-11ed-841a-96dedf69f46d"},{"id":"e68ece33-f6e0-11ea-865d-ae1031dcc090","last_name":"Özgen","first_name":"Natalie Y","full_name":"Özgen, Natalie Y"},{"first_name":"Alexis","full_name":"Ivec, Alexis","last_name":"Ivec","id":"1d144691-e8be-11ed-9b33-bdd3077fad4c"},{"first_name":"Christoph","full_name":"Bock, Christoph","last_name":"Bock"},{"last_name":"Shigemoto","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444","full_name":"Shigemoto, Ryuichi","first_name":"Ryuichi"},{"first_name":"Simon","full_name":"Hippenmeyer, Simon","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","last_name":"Hippenmeyer"}],"isi":1,"acknowledged_ssus":[{"_id":"Bio"},{"_id":"M-Shop"},{"_id":"LifeSc"},{"_id":"PreCl"}],"day":"17","das_tickbox":"1","page":"230-246.e11","volume":112,"supplementarymaterial":"yes","article_type":"original","pmid":1,"quality_controlled":"1","file":[{"date_created":"2024-02-06T13:56:15Z","access_level":"open_access","file_name":"2024_Neuron_Cheung.pdf","relation":"main_file","date_updated":"2024-02-06T13:56:15Z","checksum":"32b3788f7085cf44a84108d8faaff3ce","content_type":"application/pdf","file_id":"14944","success":1,"file_size":5942467,"creator":"dernst"}],"publication_status":"published","fulldoi":"https://doi.org/10.1016/j.neuron.2023.11.009","publication":"Neuron","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"date_created":"2023-04-27T09:41:48Z","oa":1,"date_published":"2024-01-17T00:00:00Z","intvolume":"       112","related_material":{"link":[{"url":"https://ista.ac.at/en/news/the-pedigree-of-brain-cells/","description":"News on ISTA Website","relation":"press_release"}]},"status":"public","researchdata_availability":"yes","year":"2024","issue":"2","type":"journal_article"},{"edition":"1","date_created":"2024-08-13T12:16:41Z","date_published":"2024-08-13T00:00:00Z","related_material":{"record":[{"id":"20212","status":"public","relation":"dissertation_contains"}]},"intvolume":"      2831","year":"2024","status":"public","researchdata_availability":"no","type":"book_chapter","publication_status":"published","fulldoi":"https://doi.org/10.1007/978-1-0716-3969-6_19","publication":"Neuronal Morphogenesis","volume":2831,"supplementarymaterial":"yes","editor":[{"full_name":"Toyooka, Kazuhito","first_name":"Kazuhito","last_name":"Toyooka"}],"pmid":1,"quality_controlled":"1","author":[{"full_name":"Miranda, Osvaldo","first_name":"Osvaldo","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","orcid":"0000-0001-6618-6889","last_name":"Miranda"},{"orcid":"0000-0001-8457-2572","last_name":"Cheung","id":"471195F6-F248-11E8-B48F-1D18A9856A87","first_name":"Giselle T","full_name":"Cheung, Giselle T"},{"first_name":"Simon","full_name":"Hippenmeyer, Simon","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","last_name":"Hippenmeyer"}],"acknowledged_ssus":[{"_id":"Bio"}],"day":"13","das_tickbox":"0","page":"283-299","title":"Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers","_id":"17425","publisher":"Springer Nature","date_updated":"2026-10-23T22:31:25Z","acknowledgement":"We thank all Hippenmeyer lab members for support and discussions. This work was supported by the Scientific Service Units (SSU) at ISTA through resources provided by the Imaging & Optics Facility (IOF). O.A.M was a recipient of a DOC Fellowship (26253) of the Austrian Academy of Sciences. This work was supported by ISTA institutional funds, and The Austrian Science Fund Special Research Programmes (FWF SFB F78 Neuro Stem Modulation) to S.H.","place":"New York, NY","external_id":{"pmid":["39134857"]},"alternative_title":["Methods in Molecular Biology"],"corr_author":"1","series_title":"MIMB","department":[{"_id":"GradSch"},{"_id":"SiHi"}],"article_processing_charge":"No","citation":{"chicago":"Miranda, Osvaldo, Giselle T Cheung, and Simon Hippenmeyer. “Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers.” In <i>Neuronal Morphogenesis</i>, edited by Kazuhito Toyooka, 1st ed., 2831:283–99. MIMB. New York, NY: Springer Nature, 2024. <a href=\"https://doi.org/10.1007/978-1-0716-3969-6_19\">https://doi.org/10.1007/978-1-0716-3969-6_19</a>.","short":"O. Miranda, G.T. Cheung, S. Hippenmeyer, in:, K. Toyooka (Ed.), Neuronal Morphogenesis, 1st ed., Springer Nature, New York, NY, 2024, pp. 283–299.","ama":"Miranda O, Cheung GT, Hippenmeyer S. Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers. In: Toyooka K, ed. <i>Neuronal Morphogenesis</i>. Vol 2831. 1st ed. MIMB. New York, NY: Springer Nature; 2024:283-299. doi:<a href=\"https://doi.org/10.1007/978-1-0716-3969-6_19\">10.1007/978-1-0716-3969-6_19</a>","ieee":"O. Miranda, G. T. Cheung, and S. Hippenmeyer, “Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers,” in <i>Neuronal Morphogenesis</i>, 1st ed., vol. 2831, K. Toyooka, Ed. New York, NY: Springer Nature, 2024, pp. 283–299.","ista":"Miranda O, Cheung GT, Hippenmeyer S. 2024.Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers. In: Neuronal Morphogenesis. Methods in Molecular Biology, vol. 2831, 283–299.","mla":"Miranda, Osvaldo, et al. “Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers.” <i>Neuronal Morphogenesis</i>, edited by Kazuhito Toyooka, 1st ed., vol. 2831, Springer Nature, 2024, pp. 283–99, doi:<a href=\"https://doi.org/10.1007/978-1-0716-3969-6_19\">10.1007/978-1-0716-3969-6_19</a>.","apa":"Miranda, O., Cheung, G. T., &#38; Hippenmeyer, S. (2024). Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers. In K. Toyooka (Ed.), <i>Neuronal Morphogenesis</i> (1st ed., Vol. 2831, pp. 283–299). New York, NY: Springer Nature. <a href=\"https://doi.org/10.1007/978-1-0716-3969-6_19\">https://doi.org/10.1007/978-1-0716-3969-6_19</a>"},"doi":"10.1007/978-1-0716-3969-6_19","publication_identifier":{"isbn":["9781071639689"],"eissn":["1940-6029"],"issn":["1064-3745"],"eisbn":["9781071639696"]},"abstract":[{"lang":"eng","text":"Mosaic Analysis with Double Markers (MADM) is a powerful genetic method typically used for lineage tracing and to disentangle cell autonomous and tissue-wide roles of candidate genes with single cell resolution. Given the relatively sparse labeling, depending on which of the 19 MADM chromosomes one chooses, the MADM approach represents the perfect opportunity for cell morphology analysis. Various MADM studies include reports of morphological anomalies and phenotypes in the central nervous system (CNS). MADM for any candidate gene can easily incorporate morphological analysis within the experimental workflow. Here, we describe the methods of morphological cell analysis which we developed in the course of diverse recent MADM studies. This chapter will specifically focus on methods to quantify aspects of the morphology of neurons and astrocytes within the CNS, but these methods can broadly be applied to any MADM-labeled cells throughout the entire organism. We will cover two analyses—soma volume and dendrite characterization—of physical characteristics of pyramidal neurons in the somatosensory cortex, and two analyses—volume and Sholl analysis—of astrocyte morphology."}],"project":[{"grant_number":"26253","name":"Molecular Mechanisms Regulating Cortical Neural Stem Cell Lineage Progression and Astrocyte Development","_id":"34c9fbcb-11ca-11ed-8bc3-98fa5658610d"},{"_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression","grant_number":"F7805"}],"month":"08","language":[{"iso":"eng"}],"user_id":"317138e5-6ab7-11ef-aa6d-ffef3953e345","scopus_import":"1","oa_version":"None"},{"article_number":"1133","oa_version":"Published Version","language":[{"iso":"eng"}],"month":"04","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","_id":"14783","publisher":"MDPI","date_updated":"2024-01-16T09:29:35Z","ddc":["570"],"acknowledgement":"This research was funded by grants from the European Research Council (Consolidator grant #683154) and European Union’s Horizon 2020 research and innovation program (Marie Sklodowska-Curie Innovative Training Networks, grant #722053, EU-GliaPhD) to N.R., as well as from FP7-PEOPLE Marie Curie Intra-European Fellowship for career development (grant #622289) to G.C. We thank Elena Dossi, Grégory Ghézali, and Jérémie Teillon for support with setting up the MEA system for the two-photon microscope. We would also like to thank Tayfun Palaz for their technical assistance with the EM preparations.","external_id":{"isi":["000977445700001"],"pmid":["37190042"]},"title":"Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses","article_processing_charge":"Yes","citation":{"apa":"Cheung, G. T., Chever, O., Rollenhagen, A., Quenech’du, N., Ezan, P., Lübke, J. H. R., &#38; Rouach, N. (2023). Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses. <i>Cells</i>. MDPI. <a href=\"https://doi.org/10.3390/cells12081133\">https://doi.org/10.3390/cells12081133</a>","mla":"Cheung, Giselle T., et al. “Astroglial Connexin 43 Regulates Synaptic Vesicle Release at Hippocampal Synapses.” <i>Cells</i>, vol. 12, no. 8, 1133, MDPI, 2023, doi:<a href=\"https://doi.org/10.3390/cells12081133\">10.3390/cells12081133</a>.","ista":"Cheung GT, Chever O, Rollenhagen A, Quenech’du N, Ezan P, Lübke JHR, Rouach N. 2023. Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses. Cells. 12(8), 1133.","ieee":"G. T. Cheung <i>et al.</i>, “Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses,” <i>Cells</i>, vol. 12, no. 8. MDPI, 2023.","ama":"Cheung GT, Chever O, Rollenhagen A, et al. Astroglial connexin 43 regulates synaptic vesicle release at hippocampal synapses. <i>Cells</i>. 2023;12(8). doi:<a href=\"https://doi.org/10.3390/cells12081133\">10.3390/cells12081133</a>","chicago":"Cheung, Giselle T, Oana Chever, Astrid Rollenhagen, Nicole Quenech’du, Pascal Ezan, Joachim H. R. Lübke, and Nathalie Rouach. “Astroglial Connexin 43 Regulates Synaptic Vesicle Release at Hippocampal Synapses.” <i>Cells</i>. MDPI, 2023. <a href=\"https://doi.org/10.3390/cells12081133\">https://doi.org/10.3390/cells12081133</a>.","short":"G.T. Cheung, O. Chever, A. Rollenhagen, N. Quenech’du, P. Ezan, J.H.R. Lübke, N. Rouach, Cells 12 (2023)."},"file_date_updated":"2024-01-16T09:26:52Z","doi":"10.3390/cells12081133","abstract":[{"lang":"eng","text":"Connexin 43, an astroglial gap junction protein, is enriched in perisynaptic astroglial processes and plays major roles in synaptic transmission. We have previously found that astroglial Cx43 controls synaptic glutamate levels and allows for activity-dependent glutamine release to sustain physiological synaptic transmissions and cognitiogns. However, whether Cx43 is important for the release of synaptic vesicles, which is a critical component of synaptic efficacy, remains unanswered. Here, using transgenic mice with a glial conditional knockout of Cx43 (Cx43−/−), we investigate whether and how astrocytes regulate the release of synaptic vesicles from hippocampal synapses. We report that CA1 pyramidal neurons and their synapses develop normally in the absence of astroglial Cx43. However, a significant impairment in synaptic vesicle distribution and release dynamics were observed. In particular, the FM1-43 assays performed using two-photon live imaging and combined with multi-electrode array stimulation in acute hippocampal slices, revealed a slower rate of synaptic vesicle release in Cx43−/− mice. Furthermore, paired-pulse recordings showed that synaptic vesicle release probability was also reduced and is dependent on glutamine supply via Cx43 hemichannel (HC). Taken together, we have uncovered a role for Cx43 in regulating presynaptic functions by controlling the rate and probability of synaptic vesicle release. Our findings further highlight the significance of astroglial Cx43 in synaptic transmission and efficacy."}],"publication_identifier":{"issn":["2073-4409"]},"keyword":["General Medicine"],"has_accepted_license":"1","department":[{"_id":"SiHi"}],"article_type":"original","pmid":1,"quality_controlled":"1","file":[{"file_name":"2023_Cells_Cheung.pdf","date_created":"2024-01-16T09:26:52Z","access_level":"open_access","date_updated":"2024-01-16T09:26:52Z","relation":"main_file","checksum":"6798cd75d8857976fbc58a43fd173d68","success":1,"file_id":"14808","content_type":"application/pdf","creator":"dernst","file_size":7931643}],"volume":12,"author":[{"first_name":"Giselle T","full_name":"Cheung, Giselle T","id":"471195F6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8457-2572","last_name":"Cheung"},{"last_name":"Chever","full_name":"Chever, Oana","first_name":"Oana"},{"full_name":"Rollenhagen, Astrid","first_name":"Astrid","last_name":"Rollenhagen"},{"full_name":"Quenech’du, Nicole","first_name":"Nicole","last_name":"Quenech’du"},{"full_name":"Ezan, Pascal","first_name":"Pascal","last_name":"Ezan"},{"first_name":"Joachim H. R.","full_name":"Lübke, Joachim H. R.","last_name":"Lübke"},{"full_name":"Rouach, Nathalie","first_name":"Nathalie","last_name":"Rouach"}],"isi":1,"day":"11","intvolume":"        12","status":"public","year":"2023","issue":"8","type":"journal_article","date_created":"2024-01-10T09:46:35Z","oa":1,"date_published":"2023-04-11T00:00:00Z","publication_status":"published","fulldoi":"https://doi.org/10.3390/cells12081133","publication":"Cells","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"}},{"oa":1,"date_published":"2022-02-08T00:00:00Z","date_created":"2022-02-20T23:01:30Z","type":"journal_article","intvolume":"        13","year":"2022","status":"public","publication":"Nature Communications","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication_status":"published","fulldoi":"https://doi.org/10.1038/s41467-022-28331-7","volume":13,"quality_controlled":"1","file":[{"checksum":"51d580aff2327dd957946208a9749e1a","relation":"main_file","date_updated":"2022-02-21T07:51:33Z","access_level":"open_access","date_created":"2022-02-21T07:51:33Z","file_name":"2022_NatureCommunications_Cheung.pdf","file_size":7910519,"creator":"dernst","file_id":"10777","content_type":"application/pdf","success":1}],"article_type":"original","pmid":1,"day":"08","author":[{"orcid":"0000-0001-8457-2572","last_name":"Cheung","id":"471195F6-F248-11E8-B48F-1D18A9856A87","first_name":"Giselle T","full_name":"Cheung, Giselle T"},{"full_name":"Bataveljic, Danijela","first_name":"Danijela","last_name":"Bataveljic"},{"full_name":"Visser, Josien","first_name":"Josien","last_name":"Visser"},{"last_name":"Kumar","first_name":"Naresh","full_name":"Kumar, Naresh"},{"full_name":"Moulard, Julien","first_name":"Julien","last_name":"Moulard"},{"first_name":"Glenn","full_name":"Dallérac, Glenn","last_name":"Dallérac"},{"full_name":"Mozheiko, Daria","first_name":"Daria","last_name":"Mozheiko"},{"last_name":"Rollenhagen","first_name":"Astrid","full_name":"Rollenhagen, Astrid"},{"last_name":"Ezan","full_name":"Ezan, Pascal","first_name":"Pascal"},{"last_name":"Mongin","first_name":"Cédric","full_name":"Mongin, Cédric"},{"last_name":"Chever","full_name":"Chever, Oana","first_name":"Oana"},{"last_name":"Bemelmans","full_name":"Bemelmans, Alexis Pierre","first_name":"Alexis Pierre"},{"last_name":"Lübke","full_name":"Lübke, Joachim","first_name":"Joachim"},{"full_name":"Leray, Isabelle","first_name":"Isabelle","last_name":"Leray"},{"first_name":"Nathalie","full_name":"Rouach, Nathalie","last_name":"Rouach"}],"isi":1,"title":"Physiological synaptic activity and recognition memory require astroglial glutamine","acknowledgement":"We thank D. Mazaud and. J. Cazères for technical assistance. This work was supported by grants from the European Research Council (Consolidator grant #683154) and European Union’s Horizon 2020 research and innovation program (Marie Sklodowska-Curie Innovative Training Networks, grant #722053, EU-GliaPhD) to N.R. and from FP7-PEOPLE Marie Curie Intra-European Fellowship for career development (grant #622289) to G.C.","external_id":{"pmid":["35136061"],"isi":["000757297200017"]},"publisher":"Springer Nature","_id":"10764","date_updated":"2026-04-02T12:22:38Z","ddc":["570"],"has_accepted_license":"1","department":[{"_id":"SiHi"}],"citation":{"apa":"Cheung, G. T., Bataveljic, D., Visser, J., Kumar, N., Moulard, J., Dallérac, G., … Rouach, N. (2022). Physiological synaptic activity and recognition memory require astroglial glutamine. <i>Nature Communications</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41467-022-28331-7\">https://doi.org/10.1038/s41467-022-28331-7</a>","mla":"Cheung, Giselle T., et al. “Physiological Synaptic Activity and Recognition Memory Require Astroglial Glutamine.” <i>Nature Communications</i>, vol. 13, 753, Springer Nature, 2022, doi:<a href=\"https://doi.org/10.1038/s41467-022-28331-7\">10.1038/s41467-022-28331-7</a>.","ama":"Cheung GT, Bataveljic D, Visser J, et al. Physiological synaptic activity and recognition memory require astroglial glutamine. <i>Nature Communications</i>. 2022;13. doi:<a href=\"https://doi.org/10.1038/s41467-022-28331-7\">10.1038/s41467-022-28331-7</a>","short":"G.T. Cheung, D. Bataveljic, J. Visser, N. Kumar, J. Moulard, G. Dallérac, D. Mozheiko, A. Rollenhagen, P. Ezan, C. Mongin, O. Chever, A.P. Bemelmans, J. Lübke, I. Leray, N. Rouach, Nature Communications 13 (2022).","chicago":"Cheung, Giselle T, Danijela Bataveljic, Josien Visser, Naresh Kumar, Julien Moulard, Glenn Dallérac, Daria Mozheiko, et al. “Physiological Synaptic Activity and Recognition Memory Require Astroglial Glutamine.” <i>Nature Communications</i>. Springer Nature, 2022. <a href=\"https://doi.org/10.1038/s41467-022-28331-7\">https://doi.org/10.1038/s41467-022-28331-7</a>.","ista":"Cheung GT, Bataveljic D, Visser J, Kumar N, Moulard J, Dallérac G, Mozheiko D, Rollenhagen A, Ezan P, Mongin C, Chever O, Bemelmans AP, Lübke J, Leray I, Rouach N. 2022. Physiological synaptic activity and recognition memory require astroglial glutamine. Nature Communications. 13, 753.","ieee":"G. T. Cheung <i>et al.</i>, “Physiological synaptic activity and recognition memory require astroglial glutamine,” <i>Nature Communications</i>, vol. 13. Springer Nature, 2022."},"doi":"10.1038/s41467-022-28331-7","file_date_updated":"2022-02-21T07:51:33Z","publication_identifier":{"eissn":["2041-1723"]},"abstract":[{"text":"Presynaptic glutamate replenishment is fundamental to brain function. In high activity regimes, such as epileptic episodes, this process is thought to rely on the glutamate-glutamine cycle between neurons and astrocytes. However the presence of an astroglial glutamine supply, as well as its functional relevance in vivo in the healthy brain remain controversial, partly due to a lack of tools that can directly examine glutamine transfer. Here, we generated a fluorescent probe that tracks glutamine in live cells, which provides direct visual evidence of an activity-dependent glutamine supply from astroglial networks to presynaptic structures under physiological conditions. This mobilization is mediated by connexin43, an astroglial protein with both gap-junction and hemichannel functions, and is essential for synaptic transmission and object recognition memory. Our findings uncover an indispensable recruitment of astroglial glutamine in physiological synaptic activity and memory via an unconventional pathway, thus providing an astrocyte basis for cognitive processes.","lang":"eng"}],"article_processing_charge":"No","article_number":"753","scopus_import":"1","month":"02","language":[{"iso":"eng"}],"user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","oa_version":"Published Version"},{"publication":"Journal of Visual Experiments","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication_status":"published","fulldoi":"https://doi.org/10.3791/61147","oa":1,"date_published":"2020-05-08T00:00:00Z","date_created":"2020-05-11T08:31:20Z","issue":"159","type":"journal_article","related_material":{"record":[{"relation":"part_of_dissertation","id":"7902","status":"public"}]},"year":"2020","status":"public","acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"},{"_id":"PreCl"}],"day":"08","author":[{"first_name":"Robert J","full_name":"Beattie, Robert J","last_name":"Beattie","orcid":"0000-0002-8483-8753","id":"2E26DF60-F248-11E8-B48F-1D18A9856A87"},{"id":"36BCB99C-F248-11E8-B48F-1D18A9856A87","last_name":"Streicher","first_name":"Carmen","full_name":"Streicher, Carmen"},{"full_name":"Amberg, Nicole","first_name":"Nicole","last_name":"Amberg","id":"4CD6AAC6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-3183-8207"},{"first_name":"Giselle T","full_name":"Cheung, Giselle T","id":"471195F6-F248-11E8-B48F-1D18A9856A87","last_name":"Cheung","orcid":"0000-0001-8457-2572"},{"id":"475990FE-F248-11E8-B48F-1D18A9856A87","last_name":"Contreras","full_name":"Contreras, Ximena","first_name":"Ximena"},{"first_name":"Andi H","full_name":"Hansen, Andi H","last_name":"Hansen","id":"38853E16-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Simon","full_name":"Hippenmeyer, Simon","last_name":"Hippenmeyer","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87"}],"isi":1,"ec_funded":1,"quality_controlled":"1","file":[{"date_created":"2020-05-11T08:28:38Z","access_level":"open_access","file_name":"jove-protocol-61147-lineage-tracing-clonal-analysis-developing-cerebral-cortex-using.pdf","relation":"main_file","date_updated":"2020-07-14T12:48:03Z","checksum":"3154ea7f90b9fb45e084cd1c2770597d","content_type":"application/pdf","file_id":"7816","file_size":1352186,"creator":"rbeattie"}],"article_type":"original","pmid":1,"has_accepted_license":"1","department":[{"_id":"SiHi"}],"corr_author":"1","file_date_updated":"2020-07-14T12:48:03Z","doi":"10.3791/61147","citation":{"apa":"Beattie, R. J., Streicher, C., Amberg, N., Cheung, G. T., Contreras, X., Hansen, A. H., &#38; Hippenmeyer, S. (2020). Lineage tracing and clonal analysis in developing cerebral cortex using mosaic analysis with double markers (MADM). <i>Journal of Visual Experiments</i>. MyJove Corporation. <a href=\"https://doi.org/10.3791/61147\">https://doi.org/10.3791/61147</a>","mla":"Beattie, Robert J., et al. “Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM).” <i>Journal of Visual Experiments</i>, no. 159, e61147, MyJove Corporation, 2020, doi:<a href=\"https://doi.org/10.3791/61147\">10.3791/61147</a>.","ista":"Beattie RJ, Streicher C, Amberg N, Cheung GT, Contreras X, Hansen AH, Hippenmeyer S. 2020. Lineage tracing and clonal analysis in developing cerebral cortex using mosaic analysis with double markers (MADM). Journal of Visual Experiments. (159), e61147.","ieee":"R. J. Beattie <i>et al.</i>, “Lineage tracing and clonal analysis in developing cerebral cortex using mosaic analysis with double markers (MADM),” <i>Journal of Visual Experiments</i>, no. 159. MyJove Corporation, 2020.","ama":"Beattie RJ, Streicher C, Amberg N, et al. Lineage tracing and clonal analysis in developing cerebral cortex using mosaic analysis with double markers (MADM). <i>Journal of Visual Experiments</i>. 2020;(159). doi:<a href=\"https://doi.org/10.3791/61147\">10.3791/61147</a>","chicago":"Beattie, Robert J, Carmen Streicher, Nicole Amberg, Giselle T Cheung, Ximena Contreras, Andi H Hansen, and Simon Hippenmeyer. “Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM).” <i>Journal of Visual Experiments</i>. MyJove Corporation, 2020. <a href=\"https://doi.org/10.3791/61147\">https://doi.org/10.3791/61147</a>.","short":"R.J. Beattie, C. Streicher, N. Amberg, G.T. Cheung, X. Contreras, A.H. Hansen, S. Hippenmeyer, Journal of Visual Experiments (2020)."},"publication_identifier":{"issn":["1940-087X"]},"abstract":[{"lang":"eng","text":"Beginning from a limited pool of progenitors, the mammalian cerebral cortex forms highly organized functional neural circuits. However, the underlying cellular and molecular mechanisms regulating lineage transitions of neural stem cells (NSCs) and eventual production of neurons and glia in the developing neuroepithelium remains unclear. Methods to trace NSC division patterns and map the lineage of clonally related cells have advanced dramatically. However, many contemporary lineage tracing techniques suffer from the lack of cellular resolution of progeny cell fate, which is essential for deciphering progenitor cell division patterns. Presented is a protocol using mosaic analysis with double markers (MADM) to perform in vivo clonal analysis. MADM concomitantly manipulates individual progenitor cells and visualizes precise division patterns and lineage progression at unprecedented single cell resolution. MADM-based interchromosomal recombination events during the G2-X phase of mitosis, together with temporally inducible CreERT2, provide exact information on the birth dates of clones and their division patterns. Thus, MADM lineage tracing provides unprecedented qualitative and quantitative optical readouts of the proliferation mode of stem cell progenitors at the single cell level. MADM also allows for examination of the mechanisms and functional requirements of candidate genes in NSC lineage progression. This method is unique in that comparative analysis of control and mutant subclones can be performed in the same tissue environment in vivo. Here, the protocol is described in detail, and experimental paradigms to employ MADM for clonal analysis and lineage tracing in the developing cerebral cortex are demonstrated. Importantly, this protocol can be adapted to perform MADM clonal analysis in any murine stem cell niche, as long as the CreERT2 driver is present."}],"article_processing_charge":"No","title":"Lineage tracing and clonal analysis in developing cerebral cortex using mosaic analysis with double markers (MADM)","external_id":{"pmid":["32449730"],"isi":["000546406600043"]},"_id":"7815","publisher":"MyJove Corporation","ddc":["570"],"date_updated":"2026-10-23T22:30:20Z","scopus_import":"1","month":"05","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","language":[{"iso":"eng"}],"oa_version":"Published Version","article_number":"e61147","project":[{"_id":"264E56E2-B435-11E9-9278-68D0E5697425","name":"Molecular Mechanisms Regulating Gliogenesis in the Neocortex","call_identifier":"FWF","grant_number":"M02416"},{"grant_number":"T01031","_id":"268F8446-B435-11E9-9278-68D0E5697425","name":"Role of Eed in neural stem cell lineage progression","call_identifier":"FWF"},{"grant_number":"754411","call_identifier":"H2020","_id":"260C2330-B435-11E9-9278-68D0E5697425","name":"ISTplus - Postdoctoral Fellowships"},{"name":"Molecular mechanisms of radial neuronal migration","_id":"2625A13E-B435-11E9-9278-68D0E5697425","grant_number":"24812"},{"call_identifier":"H2020","name":"Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development","_id":"260018B0-B435-11E9-9278-68D0E5697425","grant_number":"725780"}]},{"scopus_import":"1","month":"12","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","language":[{"iso":"eng"}],"oa_version":"Published Version","title":"Synaptic vesicle generation from activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction","external_id":{"isi":["000490703100001"],"pmid":["31479508"]},"_id":"7005","publisher":"Wiley","ddc":["570"],"date_updated":"2023-08-30T07:21:50Z","has_accepted_license":"1","department":[{"_id":"SiHi"}],"file_date_updated":"2020-07-14T12:47:47Z","citation":{"ista":"Cheung GT, Cousin MA. 2019. Synaptic vesicle generation from activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction. Journal of Neurochemistry. 151(5), 570–583.","ieee":"G. T. Cheung and M. A. Cousin, “Synaptic vesicle generation from activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction,” <i>Journal of Neurochemistry</i>, vol. 151, no. 5. Wiley, pp. 570–583, 2019.","ama":"Cheung GT, Cousin MA. Synaptic vesicle generation from activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction. <i>Journal of Neurochemistry</i>. 2019;151(5):570-583. doi:<a href=\"https://doi.org/10.1111/jnc.14862\">10.1111/jnc.14862</a>","short":"G.T. Cheung, M.A. Cousin, Journal of Neurochemistry 151 (2019) 570–583.","chicago":"Cheung, Giselle T, and Michael A. Cousin. “Synaptic Vesicle Generation from Activity‐dependent Bulk Endosomes Requires a Dephosphorylation‐dependent Dynamin–Syndapin Interaction.” <i>Journal of Neurochemistry</i>. Wiley, 2019. <a href=\"https://doi.org/10.1111/jnc.14862\">https://doi.org/10.1111/jnc.14862</a>.","apa":"Cheung, G. T., &#38; Cousin, M. A. (2019). Synaptic vesicle generation from activity‐dependent bulk endosomes requires a dephosphorylation‐dependent dynamin–syndapin interaction. <i>Journal of Neurochemistry</i>. Wiley. <a href=\"https://doi.org/10.1111/jnc.14862\">https://doi.org/10.1111/jnc.14862</a>","mla":"Cheung, Giselle T., and Michael A. Cousin. “Synaptic Vesicle Generation from Activity‐dependent Bulk Endosomes Requires a Dephosphorylation‐dependent Dynamin–Syndapin Interaction.” <i>Journal of Neurochemistry</i>, vol. 151, no. 5, Wiley, 2019, pp. 570–83, doi:<a href=\"https://doi.org/10.1111/jnc.14862\">10.1111/jnc.14862</a>."},"doi":"10.1111/jnc.14862","publication_identifier":{"issn":["0022-3042"],"eissn":["1471-4159"]},"abstract":[{"lang":"eng","text":"Activity-dependent bulk endocytosis generates synaptic vesicles (SVs) during intense neuronal activity via a two-step process. First, bulk endosomes are formed direct from the plasma membrane from which SVs are then generated. SV generation from bulk endosomes requires the efflux of previously accumulated calcium and activation of the protein phosphatase calcineurin. However, it is still unknown how calcineurin mediates SV generation. We addressed this question using a series of acute interventions that decoupled the generation of SVs from bulk endosomes in rat primary neuronal culture. This was achieved by either disruption of protein–protein interactions via delivery of competitive peptides, or inhibition of enzyme activity by known inhibitors. SV generation was monitored using either a morphological horseradish peroxidase assay or an optical assay that monitors the replenishment of the reserve SV pool. We found that SV generation was inhibited by, (i) peptides that disrupt calcineurin interactions, (ii) an inhibitor of dynamin I GTPase activity and (iii) peptides that disrupt the phosphorylation-dependent dynamin I–syndapin I interaction. Peptides that disrupted syndapin I interactions with eps15 homology domain-containing proteins had no effect. This revealed that (i) calcineurin must be localized at bulk endosomes to mediate its effect, (ii) dynamin I GTPase activity is essential for SV fission and (iii) the calcineurin-dependent interaction between dynamin I and syndapin I is essential for SV generation. We therefore propose that a calcineurin-dependent dephosphorylation cascade that requires both dynamin I GTPase and syndapin I lipid-deforming activity is essential for SV generation from bulk endosomes."}],"article_processing_charge":"No","volume":151,"quality_controlled":"1","file":[{"file_id":"7452","content_type":"application/pdf","creator":"dernst","file_size":4334962,"file_name":"2019_JournNeurochemistry_Cheung.pdf","access_level":"open_access","date_created":"2020-02-05T10:30:02Z","checksum":"ec1fb2aebb874009bc309adaada6e1d7","relation":"main_file","date_updated":"2020-07-14T12:47:47Z"}],"article_type":"original","pmid":1,"day":"01","author":[{"full_name":"Cheung, Giselle T","first_name":"Giselle T","orcid":"0000-0001-8457-2572","last_name":"Cheung","id":"471195F6-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Cousin","first_name":"Michael A.","full_name":"Cousin, Michael A."}],"isi":1,"page":"570-583","oa":1,"date_published":"2019-12-01T00:00:00Z","date_created":"2019-11-12T14:37:08Z","issue":"5","type":"journal_article","intvolume":"       151","year":"2019","status":"public","publication":"Journal of Neurochemistry","tmp":{"image":"/images/cc_by.png","short":"CC BY (4.0)","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode"},"publication_status":"published","fulldoi":"https://doi.org/10.1111/jnc.14862"}]