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Key roles for SRF in embryonic epidermal differentiation

ABSTRACT: During development, a polarized sheet of epidermal cells undergoes stratification and differentiation to produce the skin barrier. Through mechanisms poorly understood, the process involves adhesion and Notch signaling. To elucidate how epidermal embryogenesis is governed, we conditionally targeted transcription factor serum response factor (SRF), which has been shown to be essential for proper epidermal differentiation in vitro and in vivo. Seeking mechanism, we identified actomyosin-related genes as well-known SRF targets downregulated shortly after ablation. We show that this results in a diminished cortical actomyosin network which fails to regulate the transition of cells from the basal proliferative layer to the suprabasal differentiating layer resulting in an inability of cells to properly execute stratification and differentiation.

ORGANISM(S): Mus musculus

PROVIDER: GSE25548 | GEO | 2011/01/20

SECONDARY ACCESSION(S): PRJNA134027

REPOSITORIES: GEO

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Key roles for SRF in embryonic epidermal differentiation

Project description:During development, a polarized sheet of epidermal cells undergoes stratification and differentiation to produce the skin barrier. Through mechanisms poorly understood, the process involves adhesion and Notch signaling. To elucidate how epidermal embryogenesis is governed, we conditionally targeted transcription factor serum response factor (SRF), which has been shown to be essential for proper epidermal differentiation in vitro and in vivo. Seeking mechanism, we identified actomyosin-related genes as well-known SRF targets downregulated shortly after ablation. We show that this results in a diminished cortical actomyosin network which fails to regulate the transition of cells from the basal proliferative layer to the suprabasal differentiating layer resulting in an inability of cells to properly execute stratification and differentiation. FACS purification of basal epidermal cells from E16.5 SRF fl/fl ROSA26 YFP fl/fl K14-Cre (cKO) or SRF fl/+ ROSA26 YFP fl/fl K14-Cre (WT) was performed on a FACS Vantage SE system equipped with FACS DiVa software (BD Biosciences). Cells were gated for single events and viability and then sorted according to α6 integrin-PE expression and YFP.

2011-01-20 | E-GEOD-25548 | biostudies-arrayexpress

Mechanical Regulation of Stem Cell Proliferation and Fate Decisions by their Differentiated Daughters

Project description:RNAseq analysis of suprabasal cells in the epidermis of control and K10rtTA;TRESpastin (which depolymerize microtubules) at E16.5 were performed. We observed the premature differentiation of the barrier and increased actomyosin contractility in suprabasal differentiated cells upon microtubule disruption compared to control, and proved the increased actomyosin contractility caused by microtubule depolymerization leads to basal stem cell hyperproliferation during epidermal development.

2020-12-01 | GSE158785 | GEO

Homo sapiens

Project description:Epidermal differentiation: basal vs. suprabasal keratinocytes

| PRJNA135569 | ENA

Silencing of ultra-conserved long non-coding RNA Uc.291 causes delay of keratinocyte differentiation

Project description:Development of the skin includes a complicated program of cellular differentiation. Here, we study the role of Uc.291, a long non-coding RNA derived from ultra-conserved region, upregulated in suprabasal layer of the skin and during in vitro differentiation of keratinocytes. Depletion of Uc.291 causes delay of keratinocyte differentiation with decreased levels of the expression of epidermal differentiation complex (EDC) genes.

2020-01-08 | GSE103890 | GEO

Expression data in Srf KO and Srf WT murine muscle cells

Project description:The objective of the study is to caracherize the genes that are regulated by Srf in myoblasts at day 0 (J0) and in differentiated cells at day 1 (J1) and day 3 (J3). We used microarrays to investigate gene expression in Srf KO and Srf WT muscle cells at day 0 (J0), day 1 (J1) and day 3 (J3) of differentiation

2017-12-20 | GSE105125 | GEO

Transgenic overexpression of Pla2g2f at a level relevant to skin pathology display alopecia and psoriasis-like epidermal hyperplasia

2015-08-08 | GSE71829 | GEO

Accumulation of progerin affects the symmetry of cell division and is associated with impaired Wnt signaling and the mislocalization of nuclear envelope proteins [Affymetrix]

Project description:Hutchinson-Gilford progeria syndrome (HGPS) is the result of a defective form of the lamin A protein called progerin. While progerin is known to disrupt the properties of the nuclear lamina, the underlying mechanisms responsible for the pathophysiology of HGPS remain less clear. Previous studies in our laboratory have shown that progerin expression in murine epidermal basal cells results in impaired stratification and halted development of the skin. Stratification and differentiation of the epidermis is regulated by asymmetric stem cell division. In this dataset, we include the expression data using Affymetrix array. The samples were obtained from basal and suprabasal keratinocyes from 4-day old wild-type and bitransgenic (BT) mice.

2019-08-13 | GSE131310 | GEO

Transgenic overexpression of Pla2g2f at a level relevant to skin pathology display alopecia and psoriasis-like epidermal hyperplasia

Project description:Pla2g2f is dominantly expressed in the suprabasal layer of mouse epidermis. Microarray gene profiling supported the overall tendency of epidermal and sebaceous gland hyperplasia as well as alopecia in Pla2g2f-transgenic skin. Pla2g2f-Tg/+ mice and littermate controls (C57BL/6 background); 25-day old; skin; pooled from 4 mice for each genotype.

2015-08-08 | E-GEOD-71829 | biostudies-arrayexpress

Epidermal differentiation: basal vs. suprabasal keratinocytes

Project description:In epidermal differentiation basal keratinocytes detach from the basement membrane, stop proliferating, and express a new set of structural proteins and enzymes, which results in an impermeable protein/lipid barrier that protects us. To define the transcriptional changes essential for this process, we purified large quantities of basal and suprabasal cells from human epidermis, using the expression of b4 integrin as the discriminating factor. The expected expression differences in cytoskeletal, cell cycle and adhesion genes confirmed the effective separation of the cell populations. Using DNA microarray chips, we comprehensively identify the differences in genes expressed in basal and differentiating layers of the epidermis, including the ECM components produced by the basal cells, the proteases in both the basal and suprabasal cells, and the lipid and steroid metabolism enzymes in suprabasal cells responsible for the permeability barrier. We identified the signaling pathways specific for the two populations, and found two previously unknown paracrine and one juxtacrine signaling pathway operating between the basal and suprabasal cells. Furthermore, using specific expression signatures, we identified a new set of late differentiation markers and mapped their chromosomal loci, as well as a new set of melanocyte-specific markers. The data represent a quantum jump in understanding the mechanisms of epidermal differentiation. Basal keratinocytes are defined as integrin b4-positive. Suprabasal sample contains all melanocytes as well. Basal (integrin b4+), suprabasal (b4-) and total epidermal keratinocytes. In duplicate, from 2 independent donors, breast reduction surgery.

2010-12-15 | E-GEOD-26059 | biostudies-arrayexpress

PF4-Cre mediated deletion of SRF in murine megakaryocytes.

Project description:Srf is a MADS-box transcription factor that is critical for muscle differentiation. Its function in hematopoiesis has not yet been revealed. Mkl1, a cofactor of Srf, is part of the t(1;22) translocation in acute megakaryoblastic leukemia, and plays a critical role in megakaryopoiesis. In order to test the role of Srf in megakaryocyte development, we crossed Pf4-Cre mice, which express Cre recombinase in cells committed to the megakaryocytic lineage, to SrfF/F mice in which functional Srf is no longer expressed after Cre-mediated excision. Pf4-Cre/SrfF/F (KO) mice are born with normal mendelian frequency, but have significant macrothrombocytopenia with approximately 50% reduction in platelet count. In contrast, the BM has increased numbers and percentages of CD41+ megakaryocytes (WT: 0.41+/-0.06%; KO: 1.92+/-0.12%) with significantly reduced ploidy. KO mice show significantly increased megakaryocyte progenitors in the BM by both FACS analysis and CFU-Mk. Megakaryocytes lacking Srf have abnormal stress fiber and demarcation membrane formation and platelets lacking Srf have abnormal actin distribution. In vitro and in vivo assays reveal platelet function defects in KO mice. Critical actin cytoskeletal genes are downregulated in KO megakaryocytes. Thus, Srf is required for normal megakaryocyte maturation and platelet production, due at least in part, to regulation of cytoskeletal genes. C-kit+CD41+ megakaryocyte progenitors from PF4-Cre/SRF C57BL/6 SRF WT (3) and C57BL/6 SRF KO (3) mice were sorted by flow cytometry and cultured for three days in thrombopoietin.

2010-07-13 | E-GEOD-21859 | biostudies-arrayexpress

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