Isolation of erythroid cells from the mouse embryonic yolk sac by laser capture microdissection and subsequent microarray hybridization

Redmond, Latasha C.; Haar, Jack L.; Giebel, Mary L.; Dumur, Catherine I.; Basu, P.; Ware, Joy L.; Lloyd, Joyce A.

doi:10.1016/j.bcmd.2006.02.006

Cited by 9 publications

(15 citation statements)

References 21 publications

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“…Several global gene expression analyses derived from primary primitive or definitive erythroid cells at various stages of maturation are available (Miller, 2004; Redmond et al, 2006; Isern et al, 2011; Merryweather-Clarke et al, 2011). Of note, a comparative analysis of global gene expression in similarly staged primitive, fetal definitive, and adult definitive erythroid cells has recently been published (Kingsley et al, 2013).…”

Section: Transcriptional Regulation Of Primitive and Definitive Erythmentioning

confidence: 99%

Primitive and definitive erythropoiesis in mammals

2014

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Red blood cells (RBCs), which constitute the most abundant cell type in the body, come in two distinct flavors- primitive and definitive. Definitive RBCs in mammals circulate as smaller, anucleate cells during fetal and postnatal life, while primitive RBCs circulate transiently in the early embryo as large, nucleated cells before ultimately enucleating. Both cell types are formed from lineage-committed progenitors that generate a series of morphologically identifiable precursors that enucleate to form mature RBCs. While definitive erythroid precursors mature extravascularly in the fetal liver and postnatal marrow in association with macrophage cells, primitive erythroid precursors mature as a semi-synchronous cohort in the embryonic bloodstream. While the cytoskeletal network is critical for the maintenance of cell shape and the deformability of definitive RBCs, little is known about the components and function of the cytoskeleton in primitive erythroblasts. Erythropoietin (EPO) is a critical regulator of late-stage definitive, but not primitive, erythroid progenitor survival. However, recent studies indicate that EPO regulates multiple aspects of terminal maturation of primitive murine and human erythroid precursors, including cell survival, proliferation, and the rate of terminal maturation. Primitive and definitive erythropoiesis share central transcriptional regulators, including Gata1 and Klf1, but are also characterized by the differential expression and function of other regulators, including myb, Sox6, and Bcl11A. Flow cytometry-based methodologies, developed to purify murine and human stage-specific erythroid precursors, have enabled comparative global gene expression studies and are providing new insights into the biology of erythroid maturation.

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Section: Transcriptional Regulation Of Primitive and Definitive Erythmentioning

confidence: 99%

Primitive and definitive erythropoiesis in mammals

2014

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“…To date, the transcriptional profiles reported for erythroid cells represented only a single stage or heterogeneous populations or were derived from cells differentiated in vitro. [26][27][28][29][30] Transcription in maturing EryPs is characterized by 2 discrete waves that correlate with key developmental hallmarks. The first wave (E8.5 ϳ E9.5) coincides with the transition from YS erythropoiesis to the entry of EryPs into the bloodstream, with concomitant loss of progenitor activity.…”

Section: A Transcriptional Roadmap Of Eryps Throughout Their Developmentmentioning

confidence: 99%

Single-lineage transcriptome analysis reveals key regulatory pathways in primitive erythroid progenitors in the mouse embryo

Isern

Fraser

et al. 2011

Blood

171

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Primitive erythroid (EryP) progenitors are the first cell type specified from the mesoderm late in gastrulation. We used a transgenic reporter to image and purify the earliest blood progenitors and their descendants from developing mouse embryos. EryP progenitors exhibited remarkable proliferative capacity in the yolk sac immediately before the onset of circulation, when these cells comprise nearly half of all cells of the embryo. Global expression profiles generated at 24-hour intervals from embryonic day 7.5 through 2.5 revealed 2 abrupt changes in transcript diversity that coincided with the entry of EryPs into the circulation and with their late maturation and enucleation, respectively. These changes were paralleled by the expression of critical regulatory factors. Experiments designed to test predictions from these data demonstrated that the Wnt-signaling pathway is active in EryP progenitors, which display an aerobic glycolytic profile and the numbers of which are regulated by transforming growth factor-␤1 and hypoxia. This is the first transcriptome assembled for a single hematopoietic lineage of the embryo over the course of its differentiation.

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“…RNA extractions from the LCM-purified samples yielded 10 to 20 ng of RNA. RNA quality was assessed by capillary electrophoresis, and intact samples were processed for microarray hybridization as previously described (64).…”

Section: Animalsmentioning

confidence: 99%

“…Ϫ/Ϫ , and 3 KLF1 Ϫ/Ϫ KLF2 Ϫ/Ϫ samples were used for Affymetrix 430A 2.0 microarray assays (63)(64)(65).…”

Section: Expression Profiling Of Klf1mentioning

confidence: 99%

Krüppel-Like Factor 1 (KLF1), KLF2, and Myc Control a Regulatory Network Essential for Embryonic Erythropoiesis

Pang

Lemsaddek

Alhashem

et al. 2012

Molecular and Cellular Biology

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Isolation of erythroid cells from the mouse embryonic yolk sac by laser capture microdissection and subsequent microarray hybridization

Cited by 9 publications

References 21 publications

Primitive and definitive erythropoiesis in mammals

Primitive and definitive erythropoiesis in mammals

Single-lineage transcriptome analysis reveals key regulatory pathways in primitive erythroid progenitors in the mouse embryo

Krüppel-Like Factor 1 (KLF1), KLF2, and Myc Control a Regulatory Network Essential for Embryonic Erythropoiesis

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