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| Status |
Public on Sep 30, 2011 |
| Title |
Targeting the hemangioblast with a novel cell type-specific enhancer |
| Organism |
Gallus gallus |
| Experiment type |
Expression profiling by array
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| Summary |
Hemangioblasts are known as the common precursors for primitive hematopoietic and endothelial lineages. Their existence has been supported mainly by the observation that both cell types develop in close proximity and by in vitro differentiation and genetic studies. However, more compelling evidence will arise from tracking their cell fates using a lineage-specific marker. We report the identification of a hemangioblast-specific enhancer (Hb) located in the cis-regulatory region of chick Cerberus gene (cCer) that is able to direct the expression of enhanced green fluorescent protein (eGFP) to the precursors of yolk sac blood and endothelial cells in electroporated chick embryos. Moreover, we present the Hb-eGFP reporter as a powerful live imaging tool for visualizing hemangioblast cell fate and blood island morphogenesis. We hereby introduce the Hb enhancer as a valuable resource for genetically targeting the hemangioblast population as well as for studying the dynamics of vascular and blood cell development.
We used microarray analysis of Hb-eGFP expressing cells to verify the expression of hemangioblast-specific genes in this cell population.
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| Overall design |
In order to characterize the gene expression profile of the Hb-eGFP-positive population, we have isolated Hb-eGFP-positive cells and compared their expression profile with the control RFP-positive population. In brief, chicken embryos were electroporated at HH3 with Hb-eGFP and pCAGGS-RFP reporter constructs, observed under a fluorescence stereoscope and harvested at stage HH5-6 into three groups of four embryos each. Embryos were dissociated into single cell suspensions and the eGFP-positive and eGFP-negative/RFP-positive cell populations were sorted by Fluorescence Activated Cell Sorting (FACS). Cell populations were collected simultaneously into two different tubes, containing RNAlater (Ambion) and subsequently used for RNA extraction. Total RNA was isolated from triplicates of each population, evaluated for RNA integrity, reverse transcribed, amplified and hybridized against six Affymetrix Chicken Genome microarrays.
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| Contributor(s) |
Tavares AT |
| Citation(s) |
22204590, 29385069 |
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| Submission date |
Sep 29, 2011 |
| Last update date |
May 22, 2019 |
| Contact name |
Ana Teresa Tavares |
| E-mail(s) |
[email protected]
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| Organization name |
IGC
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| Street address |
Rua da Quinta Grande, 6
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| City |
Oeiras |
| ZIP/Postal code |
2781-901 |
| Country |
Portugal |
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| Platforms (1) |
| GPL3213 |
[Chicken] Affymetrix Chicken Genome Array |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA147765 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE32494_RAW.tar |
19.9 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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