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Blood. 2014 May 29;123(22):3466-77. doi: 10.1182/blood-2014-01-548305. Epub 2014 Mar 17.

Global transcriptome analyses of human and murine terminal erythroid differentiation.

Author information

1
Laboratory of Membrane Biology and Red Cell Physiology Laboratory, New York Blood Center, New York, NY; College of Life Science, Zhengzhou University, Zhengzhou, Henan, China;
2
Department of Pediatrics, Yale University School of Medicine, New Haven, CT;
3
Red Cell Physiology Laboratory, New York Blood Center, New York, NY;
4
Red Cell Physiology Laboratory, New York Blood Center, New York, NY; Molecular Biology Research Center, School of Biological Science and Technology and State Key Laboratory of Medical Genetics of China, Central South University, Changsha, Hunan, China; and.
5
Laboratory of Membrane Biology and.
6
Department of Pediatrics, Yale University School of Medicine, New Haven, CT; Departments of Pathology and Genetics, Yale University School of Medicine, New Haven, CT.

Abstract

We recently developed fluorescence-activated cell sorting (FACS)-based methods to purify morphologically and functionally discrete populations of cells, each representing specific stages of terminal erythroid differentiation. We used these techniques to obtain pure populations of both human and murine erythroblasts at distinct developmental stages. RNA was prepared from these cells and subjected to RNA sequencing analyses, creating unbiased, stage-specific transcriptomes. Tight clustering of transcriptomes from differing stages, even between biologically different replicates, validated the utility of the FACS-based assays. Bioinformatic analyses revealed that there were marked differences between differentiation stages, with both shared and dissimilar gene expression profiles defining each stage within transcriptional space. There were vast temporal changes in gene expression across the differentiation stages, with each stage exhibiting unique transcriptomes. Clustering and network analyses revealed that varying stage-specific patterns of expression observed across differentiation were enriched for genes of differing function. Numerous differences were present between human and murine transcriptomes, with significant variation in the global patterns of gene expression. These data provide a significant resource for studies of normal and perturbed erythropoiesis, allowing a deeper understanding of mechanisms of erythroid development in various inherited and acquired erythroid disorders.

Comment in

PMID:
24637361
PMCID:
PMC4041167
DOI:
10.1182/blood-2014-01-548305
[Indexed for MEDLINE]
Free PMC Article
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