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Gene expression in mouse hematopoietic stem and multi-potent progenitor cells with temporally defined divisional histories

ABSTRACT: Homeostatic hematopoietice stem cells (HSCs) with greater divisional history lose repopulating potential after very few cell divisions. Divisional history overrides both phenotype and immediate quiescence in determining functional activity. In GFP label retaining system GFP is progressively diluted when cells proceed through a cascade of divisions. We used a GFP label retaining system and performed microarray expression analyses to track the changes in the gene expression profile of bone marrow (BM) LSK cells that relates to divisional history during homeostasis.

ORGANISM(S): Mus musculus

PROVIDER: GSE48261 | GEO | 2014/03/20

SECONDARY ACCESSION(S): PRJNA209419

REPOSITORIES: GEO

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Gene expression in mouse hematopoietic stem and multi-potent progenitor cells with temporally defined divisional histories

Project description:Homeostatic hematopoietice stem cells (HSCs) with greater divisional history lose repopulating potential after very few cell divisions. Divisional history overrides both phenotype and immediate quiescence in determining functional activity. In GFP label retaining system GFP is progressively diluted when cells proceed through a cascade of divisions. We used a GFP label retaining system and performed microarray expression analyses to track the changes in the gene expression profile of bone marrow (BM) LSK cells that relates to divisional history during homeostasis. Chromatins are dynamically labeled in a double transgenic mouse (huCD34-tTA X Tet-O-H2B-GFP) by the expression of GFP tagged Histone2B. The expression is driven by a human CD34 promoter active specifically in all bone marrow (BM) hematopoietic stem and multi-potent progenitor cells. This is a pulse/chase Tet-off system where cells constantly label with GFP. Upon administration of Doxycyclin (Dox) to GFP labeled animals, further labeling is stopped during the chase period. With each round of cell division, the newly synthesized endogenous H2B will dilute H2B-GFP by one-half, whereas cells that don’t divide will retain GFP labeling. Therefore, the level of GFP reflects how many times cells have divided over the period of Dox treatment (chase). Animals were treated with Dox for 12 weeks starting at the age of 6-8 weeks. After chase, BM Lineage negative, Sca1+, Kit+ (LSK) cells with varying GFP levels were sorted for mRNA extraction. GFP levels were ranked from 0 to 4 based on the kinetic analysis of GFP fully labeled LSK cells and obvious peaks in the GFP histogram of LSK cells after Dox treatment. There is approximately 1-2 cell divisions between each level of GFP.

2014-03-20 | E-GEOD-48261 | biostudies-arrayexpress

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Project description:Hematopoietic stem cells (HSCs) exist in a dormant state, and progressively lose regenerative potency as they undergo successive divisions. Why this functional decline occurs and how this information is encoded is unclear. To better understand how this information is stored, we performed RNA sequencing on HSC populations differing only in their divisional history. Comparative analysis revealed that genes upregulated with divisions are enriched for lineage commitment factors, and are regulated by cell cycle-associated transcription factors, indicating that proliferation itself drives primitive hematopoietic lineage priming. In contrast, downregulated genes are associated with an HSC signature and are targeted by the Polycomb Repressive Complex 2 (PRC2). We find that Ezh2 targets HSC signature genes for repression, and a divisional history-dependent switch from Ezh1 to Ezh2 underlies HSC decline with progressive divisions. Thus cell divisions drive lineage priming and Ezh2 accumulation, which represses HSC signature genes and consolidates information on divisional history into memory.

2020-03-14 | GSE145772 | GEO

Murine Kidney Papilla Cells: Control GFP- vs. GFP+ label retaining cells. Cell populations were separated by FACS.

Project description:Transcriptional profiling of mouse kidney papilla cells comparing control GFP- cells with GFP+ labeled cells. Mouse kidney papillae were dissected. GFP- and GFP+ cell populations were isolated by FACS. The latter represent slow cycling label retaining cells (LRCs). Two-condition experiment, GFP- vs. GFP+ label retaining cells cells. Biological replicates: 5 control GFP- replicates, 5 GFP+ label retaining cell replicates.

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Project description:Label retaining and non-retaining muscle stem cells from young and aged H2B-GFP+/-;rtTA+/- were profiled by single cell RNA-seq at two timepoints

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Murine Kidney Papilla Cells: Control GFP- vs. GFP+ label retaining cells. Cell populations were separated by FACS.

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Hematopoietic stem cells in perisinusoidal niches are protected from ageing [aged, GFP-label retaining HSCs]

Project description:Hematopoietic stem cells in perisinusoidal niches are protected from ageing [aged, GFP-label retaining HSCs]

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| EGAD00010001968 | EGA

HMEC Non-label-retaining cells

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ATF4 marks quiescent reserve cells of gastric chief cells

Project description:Hierarchically organized tissues, such as hematopoietic systems, muscle, or skin harbor deeply quiescent stem cells which start proliferating in response to external insults. In contrast, it remains obscure whether similar quiescent cells exist in epithelia of digestive organs. Here we identified a deeply quiescent population in gastric corpus but not in other gastrointestinal organs after systematic examination of H2b-GFP label-retaining cells. The label-retaining cells in corpus epithelia belonged to a subpopulation of chief cells that were located near basal layers of corpus and did not overlap with Troyhigh, Lgr5high, or Misthigh cell population. The identified quiescent cells were marked with activation of Atf4 and unfolded protein response. External damages by indomethacin treatment triggered proliferation of the quiescent populations, indicating that chief cells of gastric corpus harbor deeply quiescent reserve cells with high levels of internal stress response activity.

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