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  1. Article ; Online: Protocol for sorting cells from mouse brains labeled with mosaic analysis with double markers by flow cytometry

    Nicole Amberg / Giselle Cheung / Simon Hippenmeyer

    STAR Protocols, Vol 5, Iss 1, Pp 102771- (2024)

    1481  

    Abstract: Summary: 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 ... ...

    Abstract Summary: 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.For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
    Keywords Single Cell ; Flow Cytometry ; Developmental biology ; Science (General) ; Q1-390
    Subject code 004
    Language English
    Publishing date 2024-03-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Genetic mosaic dissection of candidate genes in mice using mosaic analysis with double markers

    Nicole Amberg / Simon Hippenmeyer

    STAR Protocols, Vol 2, Iss 4, Pp 100939- (2021)

    2021  

    Abstract: Summary: Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice. MADM enables concomitant fluorescent cell labeling and introduction of a mutation of a gene of interest with single-cell resolution. ... ...

    Abstract Summary: Mosaic analysis with double markers (MADM) technology enables the generation of genetic mosaic tissue in mice. MADM enables concomitant fluorescent cell labeling and introduction of a mutation of a gene of interest with single-cell resolution. This protocol highlights major steps for the generation of genetic mosaic tissue and the isolation and processing of respective tissues for downstream histological analysis.For complete details on the use and execution of this protocol, please refer to Contreras et al. (2021).
    Keywords Single Cell ; Developmental biology ; Genetics ; Microscopy ; Model Organisms ; Molecular Biology ; Science (General) ; Q1-390
    Language English
    Publishing date 2021-12-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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

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

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 14

    Abstract: How the imprinted Cdkn1c locus regulates corticogenesis is unclear. Here, the authors dissect the level of cell-autonomy of imprinted Cdkn1c gene function in mouse corticogenesis and identify this as regulating radial glial progenitor cell and projection ...

    Abstract How the imprinted Cdkn1c locus regulates corticogenesis is unclear. Here, the authors dissect the level of cell-autonomy of imprinted Cdkn1c gene function in mouse corticogenesis and identify this as regulating radial glial progenitor cell and projection neuron survival.
    Keywords Science ; Q
    Language English
    Publishing date 2020-01-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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

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

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 14

    Abstract: How the imprinted Cdkn1c locus regulates corticogenesis is unclear. Here, the authors dissect the level of cell-autonomy of imprinted Cdkn1c gene function in mouse corticogenesis and identify this as regulating radial glial progenitor cell and projection ...

    Abstract How the imprinted Cdkn1c locus regulates corticogenesis is unclear. Here, the authors dissect the level of cell-autonomy of imprinted Cdkn1c gene function in mouse corticogenesis and identify this as regulating radial glial progenitor cell and projection neuron survival.
    Keywords Science ; Q
    Language English
    Publishing date 2020-01-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: EGFR Controls Hair Shaft Differentiation in a p53-Independent Manner

    Nicole Amberg / Panagiota A. Sotiropoulou / Gerwin Heller / Beate M. Lichtenberger / Martin Holcmann / Bahar Camurdanoglu / Temenuschka Baykuscheva-Gentscheva / Cedric Blanpain / Maria Sibilia

    iScience, Vol 15, Iss , Pp 243-

    2019  Volume 256

    Abstract: Summary: Epidermal growth factor receptor (EGFR) signaling controls skin development and homeostasis in mice and humans, and its deficiency causes severe skin inflammation, which might affect epidermal stem cell behavior. Here, we describe the ... ...

    Abstract Summary: Epidermal growth factor receptor (EGFR) signaling controls skin development and homeostasis in mice and humans, and its deficiency causes severe skin inflammation, which might affect epidermal stem cell behavior. Here, we describe the inflammation-independent effects of EGFR deficiency during skin morphogenesis and in adult hair follicle stem cells. Expression and alternative splicing analysis of RNA sequencing data from interfollicular epidermis and outer root sheath indicate that EGFR controls genes involved in epidermal differentiation and also in centrosome function, DNA damage, cell cycle, and apoptosis. Genetic experiments employing p53 deletion in EGFR-deficient epidermis reveal that EGFR signaling exhibits p53-dependent functions in proliferative epidermal compartments, as well as p53-independent functions in differentiated hair shaft keratinocytes. Loss of EGFR leads to absence of LEF1 protein specifically in the innermost epithelial hair layers, resulting in disorganization of medulla cells. Thus, our results uncover important spatial and temporal features of cell-autonomous EGFR functions in the epidermis. : Biological Sciences; Cell Biology; Developmental Biology; Stem Cells Research Subject Areas: Biological Sciences, Cell Biology, Developmental Biology, Stem Cells Research
    Keywords Science ; Q
    Subject code 571
    Language English
    Publishing date 2019-05-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: Specific roles for dendritic cell subsets during initiation and progression of psoriasis

    Elisabeth Glitzner / Ana Korosec / Patrick M Brunner / Barbara Drobits / Nicole Amberg / Helia B Schonthaler / Tamara Kopp / Erwin F Wagner / Georg Stingl / Martin Holcmann / Maria Sibilia

    EMBO Molecular Medicine, Vol 6, Iss 10, Pp 1312-

    2014  Volume 1327

    Abstract: Abstract Several subtypes of APCs are found in psoriasis patients, but their involvement in disease pathogenesis is poorly understood. Here, we investigated the contribution of Langerhans cells (LCs) and plasmacytoid DCs (pDCs) in psoriasis. In human ... ...

    Abstract Abstract Several subtypes of APCs are found in psoriasis patients, but their involvement in disease pathogenesis is poorly understood. Here, we investigated the contribution of Langerhans cells (LCs) and plasmacytoid DCs (pDCs) in psoriasis. In human psoriatic lesions and in a psoriasis mouse model (DKO* mice), LCs are severely reduced, whereas pDCs are increased. Depletion of pDCs in DKO* mice prior to psoriasis induction resulted in a milder phenotype, whereas depletion during active disease had no effect. In contrast, while depletion of Langerin‐expressing APCs before disease onset had no effect, depletion from diseased mice aggravated psoriasis symptoms. Disease aggravation was due to the absence of LCs, but not other Langerin‐expressing APCs. LCs derived from DKO* mice produced increased IL‐10 levels, suggesting an immunosuppressive function. Moreover, IL‐23 production was high in psoriatic mice and further increased in the absence of LCs. Conversely, pDC depletion resulted in reduced IL‐23 production, and therapeutic inhibition of IL‐23R signaling ameliorated disease symptoms. Therefore, LCs have an anti‐inflammatory role during active psoriatic disease, while pDCs exert an instigatory function during disease initiation.
    Keywords AP‐1 ; IL‐23 ; Langerhans cells ; plasmacytoid dendritic cells ; psoriasis ; Medicine (General) ; R5-920 ; Genetics ; QH426-470
    Language English
    Publishing date 2014-10-01T00:00:00Z
    Publisher Wiley
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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