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Title: Gene expression in mouse hematopoietic stem and multi-potent progenitor cells with temporally defined divisional histories      
dateReleased:
03-20-2014
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.
privacy:
not applicable
aggregation:
instance of dataset
ID:
E-GEOD-48261
refinement:
raw
alternateIdentifiers:
48261
keywords:
functional genomics
dateModified:
06-03-2014
availability:
available
types:
gene expression
name:
Mus musculus
ID:
A-AFFY-45
name:
Affymetrix GeneChip Mouse Genome 430 2.0 [Mouse430_2]
accessURL: https://www.ebi.ac.uk/arrayexpress/files/E-GEOD-48261/E-GEOD-48261.raw.1.zip
storedIn:
ArrayExpress
qualifier:
gzip compressed
format:
TXT
accessType:
download
authentication:
none
authorization:
none
accessURL: https://www.ebi.ac.uk/arrayexpress/files/E-GEOD-48261/E-GEOD-48261.processed.1.zip
storedIn:
ArrayExpress
qualifier:
gzip compressed
format:
TXT
accessType:
download
authentication:
none
authorization:
none
accessURL: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE48261
storedIn:
Gene Expression Omnibus
qualifier:
not compressed
format:
HTML
accessType:
landing page
primary:
true
authentication:
none
authorization:
none
abbreviation:
EBI
homePage: http://www.ebi.ac.uk/
ID:
SCR:004727
name:
European Bioinformatics Institute
homePage: https://www.ebi.ac.uk/arrayexpress/
ID:
SCR:002964
name:
ArrayExpress
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