[{"doi":"10.64898/2026.05.01.722172","article_processing_charge":"No","date_published":"2026-05-05T00:00:00Z","language":[{"iso":"eng"}],"OA_type":"green","publication":"bioRxiv","status":"public","abstract":[{"text":"The generation of faithful cell-type diversity and correct projection neuron numbers is essential for cerebral cortex development. Corticogenesis is however susceptible to genetic interference of critical signaling pathways, including mutations in Mtor/Rptor that lead to microcephaly. How the loss of Rptor/mTORC1 function affects cortical developmental programs, at single cell level, is still unknown. Here, we utilized Mosaic Analysis with Double Markers (MADM) technology to probe Rptor gene function upon sparse single cell- or global tissue-wide ablation. We found that tissue-wide effects drive the etiology of cortical microcephaly upon loss of Rptor, rather than deficits in projection neuron genesis. Conversely, Rptor function is cell-autonomously required for postnatal projection neuron survival in a highly cell-type-specific manner. Collectively, our results suggest that the fine balance of precise cell-type-specific cell-autonomous Rptor/mTORC1 function in concert with non-cell-autonomous tissue-wide effects is essential for the development of a properly-sized cerebral cortex with accurate projection neuron diversity.","lang":"eng"}],"oa_version":"Preprint","type":"preprint","author":[{"first_name":"Ana","id":"68cb85a0-39f7-11eb-9559-9aaab4f6a247","orcid":"0000-0002-5615-5277","last_name":"Villalba Requena","full_name":"Villalba Requena, Ana"},{"first_name":"Robert J","id":"2E26DF60-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-8483-8753","full_name":"Beattie, Robert J","last_name":"Beattie"},{"full_name":"Pauler, Florian","last_name":"Pauler","orcid":"0000-0002-7462-0048","first_name":"Florian","id":"48EA0138-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Streicher","full_name":"Streicher, Carmen","first_name":"Carmen","id":"36BCB99C-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Miranda, Osvaldo","last_name":"Miranda","orcid":"0000-0001-6618-6889","first_name":"Osvaldo","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425"},{"first_name":"Thomas","full_name":"Krausgruber, Thomas","last_name":"Krausgruber"},{"first_name":"Martin","full_name":"Senekowitsch, Martin","last_name":"Senekowitsch"},{"last_name":"Farlik","full_name":"Farlik, Matthias","first_name":"Matthias"},{"full_name":"Bock, Christoph","last_name":"Bock","first_name":"Christoph"},{"first_name":"Thomas","full_name":"Rülicke, Thomas","last_name":"Rülicke"},{"full_name":"Hippenmeyer, Simon","last_name":"Hippenmeyer","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","first_name":"Simon"}],"day":"05","project":[{"_id":"264E56E2-B435-11E9-9278-68D0E5697425","call_identifier":"FWF","grant_number":"M02416","name":"Molecular Mechanisms Regulating Gliogenesis in the Neocortex"},{"name":"Molecular Mechanisms of Cerebral Cortex Development","grant_number":"618444","call_identifier":"FP7","_id":"25D61E48-B435-11E9-9278-68D0E5697425"},{"grant_number":"725780","name":"Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development","_id":"260018B0-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"}],"date_updated":"2026-06-16T08:45:25Z","year":"2026","oa":1,"ddc":["570"],"acknowledgement":"We thank A. Heger (IST Austria Preclinical Facility), A. Sommer (VBCF GmbH, NGS Unit), and A.\r\nNicolas (IST Austria Lab Support Facility / Mass Spectrometry Facility) for technical support; K. Ferencak,\r\nI. Aykara, P. Hirschfeld, E. Fisher, S. Laukoter, L. Andersen for initial experiments and/or assistance; and\r\nall members of the Hippenmeyer lab for discussion. This research was supported by the Scientific Service\r\nUnits (SSU) of IST Austria through resources provided by the Imaging and Optics- (IOF), Lab Support-\r\n(LSF) and Preclinical Facilities (PCF). R.B. received support from FWF Meitner-Programm (M 2416). This\r\nwork was also supported by IST Austria institutional funds; the People Programme (Marie Curie Actions)\r\nof the European Union’s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement\r\nNo 618444 to S.H., and the European Research Council (ERC) under the European Union’s Horizon 2020\r\nresearch and innovation programme (grant agreement No 725780 LinPro) to S.H.","publication_status":"submitted","acknowledged_ssus":[{"_id":"PreCl"},{"_id":"LifeSc"},{"_id":"MassSpec"},{"_id":"Bio"}],"month":"05","department":[{"_id":"SiHi"}],"title":"Mtor/Rptor function globally prevents cortical microcephaly and cell-autonomously promotes postnatal neuron survival in cell type specific manner","main_file_link":[{"open_access":"1","url":"https://doi.org/10.64898/2026.05.01.722172"}],"date_created":"2026-06-09T08:08:18Z","has_accepted_license":"1","citation":{"short":"A. Villalba Requena, R.J. Beattie, F. Pauler, C. Streicher, O. Miranda, T. Krausgruber, M. Senekowitsch, M. Farlik, C. Bock, T. Rülicke, S. Hippenmeyer, BioRxiv (n.d.).","ista":"Villalba Requena A, Beattie RJ, Pauler F, Streicher C, Miranda O, Krausgruber T, Senekowitsch M, Farlik M, Bock C, Rülicke T, Hippenmeyer S. Mtor/Rptor function globally prevents cortical microcephaly and cell-autonomously promotes postnatal neuron survival in cell type specific manner. bioRxiv, <a href=\"https://doi.org/10.64898/2026.05.01.722172\">10.64898/2026.05.01.722172</a>.","ama":"Villalba Requena A, Beattie RJ, Pauler F, et al. Mtor/Rptor function globally prevents cortical microcephaly and cell-autonomously promotes postnatal neuron survival in cell type specific manner. <i>bioRxiv</i>. doi:<a href=\"https://doi.org/10.64898/2026.05.01.722172\">10.64898/2026.05.01.722172</a>","apa":"Villalba Requena, A., Beattie, R. J., Pauler, F., Streicher, C., Miranda, O., Krausgruber, T., … Hippenmeyer, S. (n.d.). Mtor/Rptor function globally prevents cortical microcephaly and cell-autonomously promotes postnatal neuron survival in cell type specific manner. <i>bioRxiv</i>. <a href=\"https://doi.org/10.64898/2026.05.01.722172\">https://doi.org/10.64898/2026.05.01.722172</a>","ieee":"A. Villalba Requena <i>et al.</i>, “Mtor/Rptor function globally prevents cortical microcephaly and cell-autonomously promotes postnatal neuron survival in cell type specific manner,” <i>bioRxiv</i>. .","chicago":"Villalba Requena, Ana, Robert J Beattie, Florian Pauler, Carmen Streicher, Osvaldo Miranda, Thomas Krausgruber, Martin Senekowitsch, et al. “Mtor/Rptor Function Globally Prevents Cortical Microcephaly and Cell-Autonomously Promotes Postnatal Neuron Survival in Cell Type Specific Manner.” <i>BioRxiv</i>, n.d. <a href=\"https://doi.org/10.64898/2026.05.01.722172\">https://doi.org/10.64898/2026.05.01.722172</a>.","mla":"Villalba Requena, Ana, et al. “Mtor/Rptor Function Globally Prevents Cortical Microcephaly and Cell-Autonomously Promotes Postnatal Neuron Survival in Cell Type Specific Manner.” <i>BioRxiv</i>, doi:<a href=\"https://doi.org/10.64898/2026.05.01.722172\">10.64898/2026.05.01.722172</a>."},"tmp":{"name":"Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc/4.0/legalcode","short":"CC BY-NC (4.0)","image":"/images/cc_by_nc.png"},"ec_funded":1,"OA_place":"repository","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","_id":"21962"},{"acknowledged_ssus":[{"_id":"Bio"},{"_id":"LifeSc"},{"_id":"PreCl"}],"month":"05","oa":1,"publication_status":"submitted","acknowledgement":"We thank Kay-Uwe Wagner (Wayne State University) for generously sharing Jak1/2–flox mouse lines; A.\r\nSommer (VBCF GmbH, NGS Unit) for technical support; N. Kim, V. Mick, S. Schnabl, S. Gobeil, and L.\r\nAndersen for technical assistance; all members of the Hippenmeyer lab for discussion and B. Novitch for\r\ncomments on earlier versions of the manuscript. This research was supported by the Scientific Service Units\r\n(SSU) of IST Austria through resources provided by the Imaging and Optics Facility (IOF), Lab Support-\r\n(LSF) and Preclinical Facilities (PCF). O.A.M received support from the Austrian Academy of Sciences\r\nÖAW (DOC 186584), and N.A. from FWF Elise Richter Program (Grant V1041T). This work was also\r\nsupported by IST Austria institutional funds; FWF SFB F78 (Neuro Stem Modulation) to S.H., and the\r\nEuropean Research Council (ERC) under the European Union’s Horizon 2020 research and innovation\r\nprogramme (grant agreement No 725780 LinPro) to S.H.","ddc":["570"],"year":"2026","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","_id":"21963","ec_funded":1,"OA_place":"repository","citation":{"apa":"Miranda, O., Contreras, X., Pauler, F., Davaatseren, A., Amberg, N., Streicher, C., … Hippenmeyer, S. (n.d.). Pten orchestrates neurogenic radial glia lineage progression and tunes neocortical astrocyte production. <i>bioRxiv</i>. <a href=\"https://doi.org/10.64898/2026.05.01.722191\">https://doi.org/10.64898/2026.05.01.722191</a>","ama":"Miranda O, Contreras X, Pauler F, et al. Pten orchestrates neurogenic radial glia lineage progression and tunes neocortical astrocyte production. <i>bioRxiv</i>. doi:<a href=\"https://doi.org/10.64898/2026.05.01.722191\">10.64898/2026.05.01.722191</a>","short":"O. Miranda, X. Contreras, F. Pauler, A. Davaatseren, N. Amberg, C. Streicher, A. Villalba Requena, A.-M. Heger, C. Marie, B.A. Hassan, T. Rülicke, S. Hippenmeyer, BioRxiv (n.d.).","ista":"Miranda O, Contreras X, Pauler F, Davaatseren A, Amberg N, Streicher C, Villalba Requena A, Heger A-M, Marie C, Hassan BA, Rülicke T, Hippenmeyer S. Pten orchestrates neurogenic radial glia lineage progression and tunes neocortical astrocyte production. bioRxiv, <a href=\"https://doi.org/10.64898/2026.05.01.722191\">10.64898/2026.05.01.722191</a>.","mla":"Miranda, Osvaldo, et al. “Pten Orchestrates Neurogenic Radial Glia Lineage Progression and Tunes Neocortical Astrocyte Production.” <i>BioRxiv</i>, doi:<a href=\"https://doi.org/10.64898/2026.05.01.722191\">10.64898/2026.05.01.722191</a>.","ieee":"O. Miranda <i>et al.</i>, “Pten orchestrates neurogenic radial glia lineage progression and tunes neocortical astrocyte production,” <i>bioRxiv</i>. .","chicago":"Miranda, Osvaldo, Ximena Contreras, Florian Pauler, Amarbayasgalan Davaatseren, Nicole Amberg, Carmen Streicher, Ana Villalba Requena, et al. “Pten Orchestrates Neurogenic Radial Glia Lineage Progression and Tunes Neocortical Astrocyte Production.” <i>BioRxiv</i>, n.d. <a href=\"https://doi.org/10.64898/2026.05.01.722191\">https://doi.org/10.64898/2026.05.01.722191</a>."},"has_accepted_license":"1","tmp":{"name":"Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)","legal_code_url":"https://creativecommons.org/licenses/by-nc/4.0/legalcode","short":"CC BY-NC (4.0)","image":"/images/cc_by_nc.png"},"title":"Pten orchestrates neurogenic radial glia lineage progression and tunes neocortical astrocyte production","department":[{"_id":"SiHi"},{"_id":"PreCl"},{"_id":"GradSch"}],"main_file_link":[{"open_access":"1","url":"https://doi.org/10.64898/2026.05.01.722191"}],"date_created":"2026-06-09T08:08:53Z","oa_version":"Preprint","publication":"bioRxiv","abstract":[{"lang":"eng","text":"The cerebral cortex consists of immense numbers of neuronal and glial cell-types derived from radial glial progenitor (RGP) cells. How RGPs generate appropriate quantities of distinct cortical cell-types to safeguard a brain of correct size, is not well understood. However, genetic aberration in human, including mutations in PTEN, lead to cortical malformation such as macrocephaly, albeit with unknown etiology. Here we utilized Mosaic Analysis with Double Markers (MADM)-based clonal analysis and single cell phenotyping to decipher the role of Pten in neurogenic and gliogenic RGP lineage progression during cortical ontogeny. While neurogenic RGP lineage progression and projection neuron production was moderately altered in the absence of Pten, cortical astrocyte production was drastically increased. Through genetic epistasis experiments we show that the loss of Pten uncouples astrocyte generation from essential growth factor signaling hubs, funneling into MAPK. Collectively, our results suggest that Pten regulates RGP lineage progression with distinct sequential functions in cortical projection neurogenesis and astrocyte production to ensure the emergence of a correctly-sized cerebral cortex."}],"corr_author":"1","status":"public","OA_type":"green","date_published":"2026-05-05T00:00:00Z","language":[{"iso":"eng"}],"article_processing_charge":"No","doi":"10.64898/2026.05.01.722191","date_updated":"2026-06-16T08:57:20Z","day":"05","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"},{"grant_number":"725780","name":"Principles of Neural Stem Cell Lineage Progression in Cerebral Cortex Development","_id":"260018B0-B435-11E9-9278-68D0E5697425","call_identifier":"H2020"}],"author":[{"full_name":"Miranda, Osvaldo","last_name":"Miranda","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","first_name":"Osvaldo","orcid":"0000-0001-6618-6889"},{"first_name":"Ximena","id":"475990FE-F248-11E8-B48F-1D18A9856A87","full_name":"Contreras, Ximena","last_name":"Contreras"},{"last_name":"Pauler","full_name":"Pauler, Florian","first_name":"Florian","id":"48EA0138-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-7462-0048"},{"full_name":"Davaatseren, Amarbayasgalan","last_name":"Davaatseren","id":"70ADC922-B424-11E9-99E3-BA18E6697425","first_name":"Amarbayasgalan"},{"orcid":"0000-0002-3183-8207","first_name":"Nicole","id":"4CD6AAC6-F248-11E8-B48F-1D18A9856A87","full_name":"Amberg, Nicole","last_name":"Amberg"},{"id":"36BCB99C-F248-11E8-B48F-1D18A9856A87","first_name":"Carmen","last_name":"Streicher","full_name":"Streicher, Carmen"},{"first_name":"Ana","id":"68cb85a0-39f7-11eb-9559-9aaab4f6a247","orcid":"0000-0002-5615-5277","last_name":"Villalba Requena","full_name":"Villalba Requena, Ana"},{"last_name":"Heger","full_name":"Heger, Anna-Magdalena","first_name":"Anna-Magdalena","id":"4B76FFD2-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Corentine","last_name":"Marie","full_name":"Marie, Corentine"},{"first_name":"Bassem A.","last_name":"Hassan","full_name":"Hassan, Bassem A."},{"first_name":"Thomas","full_name":"Rülicke, Thomas","last_name":"Rülicke"},{"last_name":"Hippenmeyer","full_name":"Hippenmeyer, Simon","orcid":"0000-0003-2279-1061","first_name":"Simon","id":"37B36620-F248-11E8-B48F-1D18A9856A87"}],"type":"preprint"},{"OA_place":"publisher","publication_identifier":{"isbn":["978-3-99078-063-3"],"issn":["2663-337X"]},"_id":"20212","user_id":"ba8df636-2132-11f1-aed0-ed93e2281fdd","date_created":"2025-08-22T14:07:00Z","department":[{"_id":"GradSch"},{"_id":"SiHi"}],"title":"Unraveling the role of Pten in cortical stem cell lineage progression using MADM","has_accepted_license":"1","citation":{"short":"O. Miranda, Unraveling the Role of Pten in Cortical Stem Cell Lineage Progression Using MADM, Institute of Science and Technology Austria, 2025.","ista":"Miranda O. 2025. Unraveling the role of Pten in cortical stem cell lineage progression using MADM. Institute of Science and Technology Austria.","apa":"Miranda, O. (2025). <i>Unraveling the role of Pten in cortical stem cell lineage progression using MADM</i>. Institute of Science and Technology Austria. <a href=\"https://doi.org/10.15479/AT-ISTA-20212\">https://doi.org/10.15479/AT-ISTA-20212</a>","ama":"Miranda O. Unraveling the role of Pten in cortical stem cell lineage progression using MADM. 2025. doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20212\">10.15479/AT-ISTA-20212</a>","ieee":"O. Miranda, “Unraveling the role of Pten in cortical stem cell lineage progression using MADM,” Institute of Science and Technology Austria, 2025.","chicago":"Miranda, Osvaldo. “Unraveling the Role of Pten in Cortical Stem Cell Lineage Progression Using MADM.” Institute of Science and Technology Austria, 2025. <a href=\"https://doi.org/10.15479/AT-ISTA-20212\">https://doi.org/10.15479/AT-ISTA-20212</a>.","mla":"Miranda, Osvaldo. <i>Unraveling the Role of Pten in Cortical Stem Cell Lineage Progression Using MADM</i>. Institute of Science and Technology Austria, 2025, doi:<a href=\"https://doi.org/10.15479/AT-ISTA-20212\">10.15479/AT-ISTA-20212</a>."},"acknowledgement":"I would also like to\r\nthank the Austrian Academy of Sciences for awarding me a 2-year DOC fellowship\r\n(DOC26253).","publication_status":"published","ddc":["570"],"oa":1,"alternative_title":["ISTA Thesis"],"month":"08","acknowledged_ssus":[{"_id":"Bio"},{"_id":"PreCl"}],"related_material":{"record":[{"relation":"part_of_dissertation","status":"public","id":"17425"}]},"keyword":["Pten","mtor","cortical development","MADM","Mapk"],"year":"2025","project":[{"grant_number":"26253","name":"Molecular Mechanisms Regulating Cortical Neural Stem Cell Lineage Progression and Astrocyte Development","_id":"34c9fbcb-11ca-11ed-8bc3-98fa5658610d"}],"day":"22","date_updated":"2026-08-26T22:30:03Z","degree_awarded":"PhD","type":"dissertation","supervisor":[{"last_name":"Hippenmeyer","full_name":"Hippenmeyer, Simon","id":"37B36620-F248-11E8-B48F-1D18A9856A87","first_name":"Simon","orcid":"0000-0003-2279-1061"}],"publisher":"Institute of Science and Technology Austria","author":[{"orcid":"0000-0001-6618-6889","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","first_name":"Osvaldo","full_name":"Miranda, Osvaldo","last_name":"Miranda"}],"status":"public","corr_author":"1","file":[{"relation":"source_file","embargo_to":"open_access","date_created":"2025-08-26T09:03:50Z","checksum":"3331f76bbef74ff4908e2d2c9262045c","file_size":32887334,"creator":"omiranda","file_id":"20230","file_name":"2025_MirandaRomero_OsvaldoAntonio_Thesis.docx","date_updated":"2026-08-26T22:30:02Z","access_level":"closed","content_type":"application/vnd.openxmlformats-officedocument.wordprocessingml.document"},{"embargo":"2026-08-26","relation":"main_file","date_created":"2025-08-26T09:05:55Z","checksum":"02509d50cff8e35c5bcbf71e8d658176","file_size":28636240,"creator":"omiranda","file_id":"20231","date_updated":"2026-08-26T22:30:02Z","file_name":"2025_MirandaRomero_OsvaldoAntonio_Thesis.pdf","access_level":"open_access","content_type":"application/pdf"}],"oa_version":"Published Version","doi":"10.15479/AT-ISTA-20212","article_processing_charge":"No","page":"119","file_date_updated":"2026-08-26T22:30:02Z","date_published":"2025-08-22T00:00:00Z","language":[{"iso":"eng"}]},{"place":"New York, NY","date_updated":"2026-08-26T22:30:02Z","day":"13","project":[{"_id":"34c9fbcb-11ca-11ed-8bc3-98fa5658610d","grant_number":"26253","name":"Molecular Mechanisms Regulating Cortical Neural Stem Cell Lineage Progression and Astrocyte Development"},{"_id":"059F6AB4-7A3F-11EA-A408-12923DDC885E","grant_number":"F7805","name":"Stem Cell Modulation in Neural Development and Regeneration/ P05-Molecular Mechanisms of Neural Stem Cell Lineage Progression"}],"publisher":"Springer Nature","author":[{"full_name":"Miranda, Osvaldo","last_name":"Miranda","orcid":"0000-0001-6618-6889","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","first_name":"Osvaldo"},{"last_name":"Cheung","full_name":"Cheung, Giselle T","first_name":"Giselle T","id":"471195F6-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8457-2572"},{"full_name":"Hippenmeyer, Simon","last_name":"Hippenmeyer","orcid":"0000-0003-2279-1061","id":"37B36620-F248-11E8-B48F-1D18A9856A87","first_name":"Simon"}],"type":"book_chapter","oa_version":"None","publication":"Neuronal Morphogenesis","status":"public","corr_author":"1","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."}],"series_title":"MIMB","date_published":"2024-08-13T00:00:00Z","language":[{"iso":"eng"}],"volume":2831,"article_processing_charge":"No","doi":"10.1007/978-1-0716-3969-6_19","page":"283-299","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","_id":"17425","edition":"1","publication_identifier":{"eisbn":["9781071639696"],"issn":["1064-3745"],"isbn":["9781071639689"],"eissn":["1940-6029"]},"citation":{"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.","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.","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>","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>","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>.","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.","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>."},"pmid":1,"editor":[{"full_name":"Toyooka, Kazuhito","last_name":"Toyooka","first_name":"Kazuhito"}],"quality_controlled":"1","department":[{"_id":"GradSch"},{"_id":"SiHi"}],"title":"Morphological Analysis of Neurons and Glia Using Mosaic Analysis with Double Markers","date_created":"2024-08-13T12:16:41Z","acknowledged_ssus":[{"_id":"Bio"}],"month":"08","scopus_import":"1","alternative_title":["Methods in Molecular Biology"],"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.","publication_status":"published","year":"2024","external_id":{"pmid":["39134857"]},"related_material":{"record":[{"id":"20212","relation":"dissertation_contains","status":"public"}]},"intvolume":"      2831"},{"date_created":"2019-11-10T11:23:58Z","main_file_link":[{"url":"https://doi.org/10.1101/820183","open_access":"1"}],"quality_controlled":"1","department":[{"_id":"GradSch"},{"_id":"SiHi"}],"title":"Identification of neural oscillations and epileptiform changes in human brain organoids","pmid":1,"citation":{"ieee":"R. A. Samarasinghe <i>et al.</i>, “Identification of neural oscillations and epileptiform changes in human brain organoids,” <i>Nature Neuroscience</i>, vol. 24. Springer Nature, p. 32, 2021.","chicago":"Samarasinghe, Ranmal A., Osvaldo Miranda, Jessie E. Buth, Simon Mitchell, Isabella Ferando, Momoko Watanabe, Arinnae Kurdian, et al. “Identification of Neural Oscillations and Epileptiform Changes in Human Brain Organoids.” <i>Nature Neuroscience</i>. Springer Nature, 2021. <a href=\"https://doi.org/10.1038/s41593-021-00906-5\">https://doi.org/10.1038/s41593-021-00906-5</a>.","mla":"Samarasinghe, Ranmal A., et al. “Identification of Neural Oscillations and Epileptiform Changes in Human Brain Organoids.” <i>Nature Neuroscience</i>, vol. 24, Springer Nature, 2021, p. 32, doi:<a href=\"https://doi.org/10.1038/s41593-021-00906-5\">10.1038/s41593-021-00906-5</a>.","short":"R.A. Samarasinghe, O. Miranda, J.E. Buth, S. Mitchell, I. Ferando, M. Watanabe, A. Kurdian, P. Golshani, K. Plath, W.E. Lowry, J.M. Parent, I. Mody, B.G. Novitch, Nature Neuroscience 24 (2021) 32.","ista":"Samarasinghe RA, Miranda O, Buth JE, Mitchell S, Ferando I, Watanabe M, Kurdian A, Golshani P, Plath K, Lowry WE, Parent JM, Mody I, Novitch BG. 2021. Identification of neural oscillations and epileptiform changes in human brain organoids. Nature Neuroscience. 24, 32.","apa":"Samarasinghe, R. A., Miranda, O., Buth, J. E., Mitchell, S., Ferando, I., Watanabe, M., … Novitch, B. G. (2021). Identification of neural oscillations and epileptiform changes in human brain organoids. <i>Nature Neuroscience</i>. Springer Nature. <a href=\"https://doi.org/10.1038/s41593-021-00906-5\">https://doi.org/10.1038/s41593-021-00906-5</a>","ama":"Samarasinghe RA, Miranda O, Buth JE, et al. Identification of neural oscillations and epileptiform changes in human brain organoids. <i>Nature Neuroscience</i>. 2021;24:32. doi:<a href=\"https://doi.org/10.1038/s41593-021-00906-5\">10.1038/s41593-021-00906-5</a>"},"publication_identifier":{"issn":["1097-6256"],"eissn":["1546-1726"]},"OA_place":"publisher","_id":"6995","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","intvolume":"        24","year":"2021","external_id":{"isi":["000687516300001"],"pmid":["34426698 "]},"acknowledgement":"We thank S. Butler, T. Carmichael and members of the laboratory of B.G.N. for helpful discussions and comments on the manuscript; N. Vishlaghi and F. Turcios-Hernandez for technical assistance, and J. Lee, S.-K. Lee, H. Shinagawa and K. Yoshikawa for valuable reagents. We also thank the UCLA Eli and Edythe Broad Stem Cell Research Center (BSCRC) and Intellectual and Developmental Disabilities Research Center microscopy cores for access to imaging facilities. This work was supported by grants from the California Institute for Regenerative Medicine (CIRM) (DISC1-08819 to B.G.N.), the National Institute of Health (R01NS089817, R01DA051897 and P50HD103557 to B.G.N.; K08NS119747 to R.A.S.; K99HD096105 to M.W.; R01MH123922, R01MH121521 and P50HD103557 to M.J.G.; R01GM099134 to K.P.; R01NS103788 to W.E.L.; R01NS088571 to J.M.P.; R01NS030549 and R01AG050474 to I.M.), and research awards from the UCLA Jonsson Comprehensive Cancer Center and BSCRC Ablon Scholars Program (to B.G.N.), the BSCRC Innovation Program (to B.G.N., K.P. and W.E.L.), the UCLA BSCRC Steffy Brain Aging Research Fund (to B.G.N. and W.E.L.) and the UCLA Clinical and Translational Science Institute (to B.G.N.), Paul Allen Family Foundation Frontiers Group (to K.P. and W.E.L.), the March of Dimes Foundation (to W.E.L.) and the Simons Foundation Autism Research Initiative Bridge to Independence Program (to R.A.S. and M.J.G.). R.A.S. was also supported by the UCLA/NINDS Translational Neuroscience Training Grant (R25NS065723), a Research and Training Fellowship from the American Epilepsy Society, a Taking Flight Award from CURE Epilepsy and a Clinician Scientist training award from the UCLA BSCRC. J.E.B. was supported by the UCLA BSCRC Rose Hills Foundation Graduate Scholarship Training Program. M.W. was supported by postdoctoral training awards provided by the UCLA BSCRC and the Uehara Memorial Foundation. O.A.M. and A.K. were supported in part by the UCLA-California State University Northridge CIRM-Bridges training program (EDUC2-08411). We also acknowledge the support of the IDDRC Cells, Circuits and Systems Analysis, Microscopy and Genetics and Genomics Cores of the Semel Institute of Neuroscience at UCLA, which are supported by the NICHD (U54HD087101 and P50HD10355701). We lastly acknowledge support from a Quantitative and Computational Biosciences Collaboratory Postdoctoral Fellowship to S.M. and the Quantitative and Computational Biosciences Collaboratory community, directed by M. Pellegrini.","publication_status":"published","article_type":"review","oa":1,"scopus_import":"1","month":"08","type":"journal_article","author":[{"first_name":"Ranmal A.","full_name":"Samarasinghe, Ranmal A.","last_name":"Samarasinghe"},{"orcid":"0000-0001-6618-6889","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","first_name":"Osvaldo","full_name":"Miranda, Osvaldo","last_name":"Miranda"},{"last_name":"Buth","full_name":"Buth, Jessie E.","first_name":"Jessie E."},{"first_name":"Simon","full_name":"Mitchell, Simon","last_name":"Mitchell"},{"first_name":"Isabella","last_name":"Ferando","full_name":"Ferando, Isabella"},{"last_name":"Watanabe","full_name":"Watanabe, Momoko","first_name":"Momoko"},{"last_name":"Kurdian","full_name":"Kurdian, Arinnae","first_name":"Arinnae"},{"first_name":"Peyman","full_name":"Golshani, Peyman","last_name":"Golshani"},{"full_name":"Plath, Kathrin","last_name":"Plath","first_name":"Kathrin"},{"first_name":"William E.","last_name":"Lowry","full_name":"Lowry, William E."},{"last_name":"Parent","full_name":"Parent, Jack M.","first_name":"Jack M."},{"first_name":"Istvan","last_name":"Mody","full_name":"Mody, Istvan"},{"full_name":"Novitch, Bennett G.","last_name":"Novitch","first_name":"Bennett G."}],"publisher":"Springer Nature","isi":1,"day":"23","date_updated":"2025-07-09T09:00:12Z","article_processing_charge":"No","doi":"10.1038/s41593-021-00906-5","page":"32","volume":24,"language":[{"iso":"eng"}],"date_published":"2021-08-23T00:00:00Z","OA_type":"green","status":"public","abstract":[{"lang":"eng","text":"Human brain organoids represent a powerful tool for the study of human neurological diseases particularly those that impact brain growth and structure. However, many neurological diseases lack obvious anatomical abnormalities, yet significantly impact neural network functions, raising the question of whether organoids possess sufficient neural network architecture and complexity to model these conditions. Here, we explore the network level functions of brain organoids using calcium sensor imaging and extracellular recording approaches that together reveal the existence of complex oscillatory network behaviors reminiscent of intact brain preparations. We further demonstrate strikingly abnormal epileptiform network activity in organoids derived from a Rett Syndrome patient despite only modest anatomical differences from isogenically matched controls, and rescue with an unconventional neuromodulatory drug Pifithrin-α. Together, these findings provide an essential foundation for the utilization of human brain organoids to study intact and disordered human brain network formation and illustrate their utility in therapeutic discovery."}],"publication":"Nature Neuroscience","oa_version":"Preprint"},{"page":"75","article_processing_charge":"No","doi":"10.1101/2019.12.13.875773","year":"2019","language":[{"iso":"eng"}],"date_published":"2019-12-13T00:00:00Z","abstract":[{"text":"Telencephalic organoids generated from human pluripotent stem cells (hPSCs) are emerging as an effective system to study the distinct features of the developing human brain and the underlying causes of many neurological disorders. While progress in organoid technology has been steadily advancing, many challenges remain including rampant batch-to-batch and cell line-to-cell line variability and irreproducibility. Here, we demonstrate that a major contributor to successful cortical organoid production is the manner in which hPSCs are maintained prior to differentiation. Optimal results were achieved using fibroblast-feeder-supported hPSCs compared to feeder-independent cells, related to differences in their transcriptomic states. Feeder-supported hPSCs display elevated activation of diverse TGFβ superfamily signaling pathways and increased expression of genes associated with naïve pluripotency. We further identify combinations of TGFβ-related growth factors that are necessary and together sufficient to impart broad telencephalic organoid competency to feeder-free hPSCs and enable reproducible formation of brain structures suitable for disease modeling.","lang":"eng"}],"status":"public","publication_status":"published","oa":1,"publication":"bioRxiv","month":"12","oa_version":"Preprint","type":"preprint","date_created":"2020-01-23T09:53:40Z","main_file_link":[{"url":"https://doi.org/10.1101/2019.12.13.875773","open_access":"1"}],"extern":"1","title":"TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids","citation":{"apa":"Watanabe, M., Haney, J. R., Vishlaghi, N., Turcios, F., Buth, J. E., Gu, W., … Novitch, B. G. (2019). TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. <i>bioRxiv</i>. Cold Spring Harbor Laboratory. <a href=\"https://doi.org/10.1101/2019.12.13.875773\">https://doi.org/10.1101/2019.12.13.875773</a>","ama":"Watanabe M, Haney JR, Vishlaghi N, et al. TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. <i>bioRxiv</i>. 2019. doi:<a href=\"https://doi.org/10.1101/2019.12.13.875773\">10.1101/2019.12.13.875773</a>","short":"M. Watanabe, J.R. Haney, N. Vishlaghi, F. Turcios, J.E. Buth, W. Gu, A.J. Collier, O. Miranda, D. Chen, S. Sabri, A.T. Clark, K. Plath, H.R. Christofk, M.J. Gandal, B.G. Novitch, BioRxiv (2019).","ista":"Watanabe M, Haney JR, Vishlaghi N, Turcios F, Buth JE, Gu W, Collier AJ, Miranda O, Chen D, Sabri S, Clark AT, Plath K, Christofk HR, Gandal MJ, Novitch BG. 2019. TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids. bioRxiv, <a href=\"https://doi.org/10.1101/2019.12.13.875773\">10.1101/2019.12.13.875773</a>.","mla":"Watanabe, Momoko, et al. “TGFβ Superfamily Signaling Regulates the State of Human Stem Cell Pluripotency and Competency to Create Telencephalic Organoids.” <i>BioRxiv</i>, Cold Spring Harbor Laboratory, 2019, doi:<a href=\"https://doi.org/10.1101/2019.12.13.875773\">10.1101/2019.12.13.875773</a>.","ieee":"M. Watanabe <i>et al.</i>, “TGFβ superfamily signaling regulates the state of human stem cell pluripotency and competency to create telencephalic organoids,” <i>bioRxiv</i>. Cold Spring Harbor Laboratory, 2019.","chicago":"Watanabe, Momoko, Jillian R. Haney, Neda Vishlaghi, Felix Turcios, Jessie E. Buth, Wen Gu, Amanda J. Collier, et al. “TGFβ Superfamily Signaling Regulates the State of Human Stem Cell Pluripotency and Competency to Create Telencephalic Organoids.” <i>BioRxiv</i>. Cold Spring Harbor Laboratory, 2019. <a href=\"https://doi.org/10.1101/2019.12.13.875773\">https://doi.org/10.1101/2019.12.13.875773</a>."},"author":[{"last_name":"Watanabe","full_name":"Watanabe, Momoko","first_name":"Momoko"},{"first_name":"Jillian R.","full_name":"Haney, Jillian R.","last_name":"Haney"},{"full_name":"Vishlaghi, Neda","last_name":"Vishlaghi","first_name":"Neda"},{"first_name":"Felix","full_name":"Turcios, Felix","last_name":"Turcios"},{"full_name":"Buth, Jessie E.","last_name":"Buth","first_name":"Jessie E."},{"first_name":"Wen","full_name":"Gu, Wen","last_name":"Gu"},{"full_name":"Collier, Amanda J.","last_name":"Collier","first_name":"Amanda J."},{"full_name":"Miranda, Osvaldo","last_name":"Miranda","orcid":"0000-0001-6618-6889","id":"862A3C56-A8BF-11E9-B4FA-D9E3E5697425","first_name":"Osvaldo"},{"first_name":"Di","last_name":"Chen","full_name":"Chen, Di"},{"last_name":"Sabri","full_name":"Sabri, Shan","first_name":"Shan"},{"last_name":"Clark","full_name":"Clark, Amander T.","first_name":"Amander T."},{"last_name":"Plath","full_name":"Plath, Kathrin","first_name":"Kathrin"},{"first_name":"Heather R.","last_name":"Christofk","full_name":"Christofk, Heather R."},{"first_name":"Michael J.","last_name":"Gandal","full_name":"Gandal, Michael J."},{"full_name":"Novitch, Bennett G.","last_name":"Novitch","first_name":"Bennett G."}],"publisher":"Cold Spring Harbor Laboratory","day":"13","_id":"7358","date_updated":"2022-06-17T08:03:32Z","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87"}]