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Article ; Online: Tissue-wide genetic and cellular landscape shapes the execution of sequential PRC2 functions in neural stem cell lineage progression.

Amberg, Nicole / Pauler, Florian M / Streicher, Carmen / Hippenmeyer, Simon

2022 Volume 8, Issue 44, Page(s) eabq1263

Abstract: The generation of a correctly sized cerebral cortex with all-embracing neuronal and glial cell-type diversity critically depends on faithful radial glial progenitor (RGP) cell proliferation/differentiation programs. Temporal RGP lineage progression is ... ...

Abstract	The generation of a correctly sized cerebral cortex with all-embracing neuronal and glial cell-type diversity critically depends on faithful radial glial progenitor (RGP) cell proliferation/differentiation programs. Temporal RGP lineage progression is regulated by Polycomb repressive complex 2 (PRC2), and loss of PRC2 activity results in severe neurogenesis defects and microcephaly. How PRC2-dependent gene expression instructs RGP lineage progression is unknown. Here, we use mosaic analysis with double markers (MADM)-based single-cell technology and demonstrate that PRC2 is not cell-autonomously required in neurogenic RGPs but rather acts at the global tissue-wide level. Conversely, cortical astrocyte production and maturation is cell-autonomously controlled by PRC2-dependent transcriptional regulation. We thus reveal highly distinct and sequential PRC2 functions in RGP lineage progression that are dependent on complex interplays between intrinsic and tissue-wide properties. In a broader context, our results imply a critical role for the genetic and cellular niche environment in neural stem cell behavior.
MeSH term(s)	Polycomb Repressive Complex 2/genetics ; Polycomb Repressive Complex 2/metabolism ; Cell Differentiation/genetics ; Neural Stem Cells ; Neurogenesis/genetics ; Neurons/metabolism
Chemical Substances	Polycomb Repressive Complex 2 (EC 2.1.1.43)
Language	English
Publishing date	2022-11-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	2810933-8
ISSN	2375-2548 ; 2375-2548
ISSN (online)	2375-2548
ISSN	2375-2548
DOI	10.1126/sciadv.abq1263
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Inducible uniparental chromosome disomy to probe genomic imprinting at single-cell level in brain and beyond.

Pauler, Florian M / Hudson, Quanah J / Laukoter, Susanne / Hippenmeyer, Simon

Neurochemistry international

2021 Volume 145, Page(s) 104986

Abstract: Genomic imprinting is an epigenetic mechanism that results in parental allele-specific expression of ~1% of all genes in mouse and human. Imprinted genes are key developmental regulators and play pivotal roles in many biological processes such as ... ...

Abstract	Genomic imprinting is an epigenetic mechanism that results in parental allele-specific expression of ~1% of all genes in mouse and human. Imprinted genes are key developmental regulators and play pivotal roles in many biological processes such as nutrient transfer from the mother to offspring and neuronal development. Imprinted genes are also involved in human disease, including neurodevelopmental disorders, and often occur in clusters that are regulated by a common imprint control region (ICR). In extra-embryonic tissues ICRs can act over large distances, with the largest surrounding Igf2r spanning over 10 million base-pairs. Besides classical imprinted expression that shows near exclusive maternal or paternal expression, widespread biased imprinted expression has been identified mainly in brain. In this review we discuss recent developments mapping cell type specific imprinted expression in extra-embryonic tissues and neocortex in the mouse. We highlight the advantages of using an inducible uniparental chromosome disomy (UPD) system to generate cells carrying either two maternal or two paternal copies of a specific chromosome to analyze the functional consequences of genomic imprinting. Mosaic Analysis with Double Markers (MADM) allows fluorescent labeling and concomitant induction of UPD sparsely in specific cell types, and thus to over-express or suppress all imprinted genes on that chromosome. To illustrate the utility of this technique, we explain how MADM-induced UPD revealed new insights about the function of the well-studied Cdkn1c imprinted gene, and how MADM-induced UPDs led to identification of highly cell type specific phenotypes related to perturbed imprinted expression in the mouse neocortex. Finally, we give an outlook on how MADM could be used to probe cell type specific imprinted expression in other tissues in mouse, particularly in extra-embryonic tissues.
MeSH term(s)	Animals ; Brain/cytology ; Brain/physiology ; Cyclin-Dependent Kinase Inhibitor p57/genetics ; Epigenesis, Genetic/physiology ; Genomic Imprinting/physiology ; Humans ; Receptor, IGF Type 2/genetics ; Single-Cell Analysis/methods ; Uniparental Disomy/genetics
Chemical Substances	Cyclin-Dependent Kinase Inhibitor p57 ; Igf2r protein, mouse ; Receptor, IGF Type 2
Language	English
Publishing date	2021-02-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	283190-9
ISSN	1872-9754 ; 0197-0186
ISSN (online)	1872-9754
ISSN	0197-0186
DOI	10.1016/j.neuint.2021.104986
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Article ; Online: Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy.

Laukoter, Susanne / Amberg, Nicole / Pauler, Florian M / Hippenmeyer, Simon

STAR protocols

2020 Volume 1, Issue 3, Page(s) 100215

Abstract: Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not ... ...

Abstract	Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments. For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b).
MeSH term(s)	Animals ; Astrocytes ; Biomarkers ; Brain/cytology ; Cell Separation/methods ; Chromosomes ; Flow Cytometry/methods ; Fluorescent Antibody Technique/methods ; Genomic Imprinting ; Mice ; Mosaicism ; Phenotype ; Single-Cell Analysis/methods ; Software ; Uniparental Disomy/cytology ; Exome Sequencing
Chemical Substances	Biomarkers
Language	English
Publishing date	2020-12-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2020.100215
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Article ; Online: SCOPES

Robert Beattie / Simon Hippenmeyer / Florian M. Pauler

Frontiers in Education, Vol

Sparking Curiosity Through Open-Source Platforms in Education and Science

2020 Volume 5

Abstract: Scientific research is to date largely restricted to wealthy laboratories in developed nations due to the necessity of complex and expensive equipment. This inequality limits the capacity of science to be used as a diplomatic channel. Maker movements use ...

Abstract	Scientific research is to date largely restricted to wealthy laboratories in developed nations due to the necessity of complex and expensive equipment. This inequality limits the capacity of science to be used as a diplomatic channel. Maker movements use open-source technologies including additive manufacturing (3D printing) and laser cutting, together with low-cost computers for developing novel products. This movement is setting the groundwork for a revolution, allowing scientific equipment to be sourced at a fraction of the cost and has the potential to increase the availability of equipment for scientists around the world. Science education is increasingly recognized as another channel for science diplomacy. In this perspective, we introduce the idea that the Maker movement and open-source technologies have the potential to revolutionize science, technology, engineering and mathematics (STEM) education worldwide. We present an open-source STEM didactic tool called SCOPES (Sparking Curiosity through Open-source Platforms in Education and Science). SCOPES is self-contained, independent of local resources, and cost-effective. SCOPES can be adapted to communicate complex subjects from genetics to neurobiology, perform real-world biological experiments and explore digitized scientific samples. We envision such platforms will enhance science diplomacy by providing a means for scientists to share their findings with classrooms and for educators to incorporate didactic concepts into STEM lessons. By providing students the opportunity to design, perform, and share scientific experiments, students also experience firsthand the benefits of a multinational scientific community. We provide instructions on how to build and use SCOPES on our webpage: http://scopeseducation.org.
Keywords	science diplomacy ; open science ; STEM ; science education ; mosaic analysis with double markers (MADM) ; neuroscience ; Education (General) ; L7-991
Subject code	028
Language	English
Publishing date	2020-05-01T00:00:00Z
Publisher	Frontiers Media S.A.
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Simultaneous brain cell type and lineage determined by scRNA-seq reveals stereotyped cortical development.

Anderson, Donovan J / Pauler, Florian M / McKenna, Aaron / Shendure, Jay / Hippenmeyer, Simon / Horwitz, Marshall S

Cell systems

2022 Volume 13, Issue 6, Page(s) 438–453.e5

Abstract: Mutations are acquired frequently, such that each cell's genome inscribes its history of cell divisions. Common genomic alterations involve loss of heterozygosity (LOH). LOH accumulates throughout the genome, offering large encoding capacity for ... ...

Abstract	Mutations are acquired frequently, such that each cell's genome inscribes its history of cell divisions. Common genomic alterations involve loss of heterozygosity (LOH). LOH accumulates throughout the genome, offering large encoding capacity for inferring cell lineage. Using only single-cell RNA sequencing (scRNA-seq) of mouse brain cells, we found that LOH events spanning multiple genes are revealed as tracts of monoallelically expressed, constitutionally heterozygous single-nucleotide variants (SNVs). We simultaneously inferred cell lineage and marked developmental time points based on X chromosome inactivation and the total number of LOH events while identifying cell types from gene expression patterns. Our results are consistent with progenitor cells giving rise to multiple cortical cell types through stereotyped expansion and distinct waves of neurogenesis. This type of retrospective analysis could be incorporated into scRNA-seq pipelines and, compared with experimental approaches for determining lineage in model organisms, is applicable where genetic engineering is prohibited, such as humans.
MeSH term(s)	Animals ; Brain ; Loss of Heterozygosity ; Mice ; Neurogenesis ; Retrospective Studies ; Single-Cell Analysis/methods
Language	English
Publishing date	2022-04-21
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2854138-8
ISSN	2405-4720 ; 2405-4712
ISSN (online)	2405-4720
ISSN	2405-4712
DOI	10.1016/j.cels.2022.03.006
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Multipotent progenitors instruct ontogeny of the superior colliculus.

Cheung, Giselle / Pauler, Florian M / Koppensteiner, Peter / Krausgruber, Thomas / Streicher, Carmen / Schrammel, Martin / Gutmann-Özgen, Natalie / Ivec, Alexis E / Bock, Christoph / Shigemoto, Ryuichi / Hippenmeyer, Simon

Neuron

2023 Volume 112, Issue 2, Page(s) 230–246.e11

Abstract: 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- ... ...

Abstract	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.
MeSH term(s)	Animals ; Superior Colliculi/physiology ; Neurons/metabolism ; Neuroglia/metabolism ; Neural Stem Cells/metabolism ; Cell Lineage/physiology ; Mammals
Language	English
Publishing date	2023-12-13
Publishing country	United States
Document type	Journal Article
ZDB-ID	808167-0
ISSN	1097-4199 ; 0896-6273
ISSN (online)	1097-4199
ISSN	0896-6273
DOI	10.1016/j.neuron.2023.11.009
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Zs.A 2496: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 92: Show issues

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Article ; Online: Imprinted Cdkn1c genomic locus cell-autonomously promotes cell survival in cerebral cortex development.

Laukoter, Susanne / Beattie, Robert / Pauler, Florian M / Amberg, Nicole / Nakayama, Keiichi I / Hippenmeyer, Simon

Nature communications

2020 Volume 11, Issue 1, Page(s) 195

Abstract: The cyclin-dependent kinase inhibitor ... ...

Abstract	The cyclin-dependent kinase inhibitor p57
MeSH term(s)	Animals ; Cell Survival ; Cerebral Cortex/growth & development ; Cerebral Cortex/metabolism ; Cyclin-Dependent Kinase Inhibitor p57/genetics ; Cyclin-Dependent Kinase Inhibitor p57/metabolism ; Female ; Gene Expression Regulation, Developmental ; Genomics ; Male ; Mice ; Mice, Knockout ; Neurogenesis/genetics ; Neurogenesis/physiology ; Neurons/classification ; Neurons/metabolism ; Phenotype ; Transcriptome
Chemical Substances	Cdkn1c protein, mouse ; Cyclin-Dependent Kinase Inhibitor p57
Language	English
Publishing date	2020-01-10
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-019-14077-2
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Generation and isolation of single cells from mouse brain with mosaic analysis with double markers-induced uniparental chromosome disomy

Susanne Laukoter / Nicole Amberg / Florian M. Pauler / Simon Hippenmeyer

STAR Protocols, Vol 1, Iss 3, Pp 100215- (2020)

2020

Abstract: Summary: Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are ... ...

Abstract	Summary: Mosaic analysis with double markers (MADM) technology enables concomitant fluorescent cell labeling and induction of uniparental chromosome disomy (UPD) with single-cell resolution. In UPD, imprinted genes are either overexpressed 2-fold or are not expressed. Here, the MADM platform is utilized to probe imprinting phenotypes at the transcriptional level. This protocol highlights major steps for the generation and isolation of projection neurons and astrocytes with MADM-induced UPD from mouse cerebral cortex for downstream single-cell and low-input sample RNA-sequencing experiments.For complete details on the use and execution of this protocol, please refer to Laukoter et al. (2020b).
Keywords	Single Cell ; Flow Cytometry/Mass Cytometry ; Genetics ; Neuroscience ; Science (General) ; Q1-390
Language	English
Publishing date	2020-12-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: LINC01133

Yotova, Iveta / Hudson, Quanah J / Pauler, Florian M / Proestling, Katharina / Haslinger, Isabella / Kuessel, Lorenz / Perricos, Alexandra / Husslein, Heinrich / Wenzl, René

International journal of molecular sciences

2021 Volume 22, Issue 16

Abstract: Endometriosis is a common gynecological disorder characterized by ectopic growth of endometrium outside the uterus and is associated with chronic pain and infertility. We investigated the role of the long intergenic noncoding RNA 01133 ( ...

Abstract	Endometriosis is a common gynecological disorder characterized by ectopic growth of endometrium outside the uterus and is associated with chronic pain and infertility. We investigated the role of the long intergenic noncoding RNA 01133 (
Language	English
Publishing date	2021-08-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms22168385
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Article ; Online: Silencing by the imprinted Airn macro lncRNA: transcription is the answer.

Santoro, Federica / Pauler, Florian M

Cell cycle (Georgetown, Tex.)

2013 Volume 12, Issue 5, Page(s) 711–712

MeSH term(s)	Animals ; Female ; Gene Silencing ; Genomic Imprinting ; Humans ; Male ; Mammals/genetics ; Models, Genetic ; Promoter Regions, Genetic/genetics ; RNA, Long Noncoding/genetics ; Receptor, IGF Type 2/genetics ; Transcription, Genetic
Chemical Substances	RNA, Long Noncoding ; Receptor, IGF Type 2
Language	English
Publishing date	2013-02-19
Publishing country	United States
Document type	Editorial
ZDB-ID	2146183-1
ISSN	1551-4005 ; 1538-4101 ; 1554-8627
ISSN (online)	1551-4005
ISSN	1538-4101 ; 1554-8627
DOI	10.4161/cc.23860
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Zs.A 5881: Show issues

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ab Jg. 2022: Lesesaal (EG)

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