Dose-dependent regulation of primitive erythroid maturation and identity by the transcription factor Eklf

Joan Isern, Stuart T. Fraser, Zhiyong He, Hailan Zhang, Margaret H. Baron

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The primitive erythroid (EryP) lineage is the first to differentiate during mammalian embryogenesis. Eklf/Klf1 is a transcriptional regulator that is essential for definitive erythropoiesis in the fetal liver. Dissection of the role(s) of Eklf within the EryP compartment has been confounded by the simultaneous presence of EryP and fetal liver-derived definitive erythroid (EryD) cells in the blood. To address this problem, we have distinguished EryP from their definitive counterparts by crossing Eklf+/- mutant and ε-globin:: histone H2B-GFP transgenic mice. Eklfdeficient EryP exhibit membrane ruffling and a failure to acquire the typical discoidal erythroid shape but they can enucleate. Flow cytometric analyses of H2B-GFP+ EryP revealed that Eklf heterozygosity results in the loss of Ter119 surface expression on EryP but not on EryD. Null mutation of Eklf resulted in abnormal expression of a range of surface proteins by EryP. In particular, several megakaryocyte markers were ectopically expressed by maturing Eklf-null EryP. Unexpectedly, the platelet tetraspanin CD9 was detected on nucleated wild-type EryP but not on mature EryD and thus provides a useful marker for purifying circulating EryP. We conclude that Eklf gene dosage is crucial for regulating the surface phenotype and molecular identity of maturing primitive erythroid cells.

Original languageEnglish (US)
Pages (from-to)3972-3980
Number of pages9
JournalBlood
Volume116
Issue number19
DOIs
StatePublished - Nov 11 2010
Externally publishedYes

Fingerprint

Erythroid Cells
Liver
Transcription Factors
Dissection
Megakaryocytes
Gene Dosage
Globins
Erythropoiesis
Platelets
Histones
Transgenic Mice
Embryonic Development
Membrane Proteins
Blood
Blood Platelets
Genes
Membranes
Phenotype
Mutation

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

Cite this

Dose-dependent regulation of primitive erythroid maturation and identity by the transcription factor Eklf. / Isern, Joan; Fraser, Stuart T.; He, Zhiyong; Zhang, Hailan; Baron, Margaret H.

In: Blood, Vol. 116, No. 19, 11.11.2010, p. 3972-3980.

Research output: Contribution to journalArticle

Isern, Joan ; Fraser, Stuart T. ; He, Zhiyong ; Zhang, Hailan ; Baron, Margaret H. / Dose-dependent regulation of primitive erythroid maturation and identity by the transcription factor Eklf. In: Blood. 2010 ; Vol. 116, No. 19. pp. 3972-3980.
@article{7579e0d612d047a784276d703d7a4bb2,
title = "Dose-dependent regulation of primitive erythroid maturation and identity by the transcription factor Eklf",
abstract = "The primitive erythroid (EryP) lineage is the first to differentiate during mammalian embryogenesis. Eklf/Klf1 is a transcriptional regulator that is essential for definitive erythropoiesis in the fetal liver. Dissection of the role(s) of Eklf within the EryP compartment has been confounded by the simultaneous presence of EryP and fetal liver-derived definitive erythroid (EryD) cells in the blood. To address this problem, we have distinguished EryP from their definitive counterparts by crossing Eklf+/- mutant and ε-globin:: histone H2B-GFP transgenic mice. Eklfdeficient EryP exhibit membrane ruffling and a failure to acquire the typical discoidal erythroid shape but they can enucleate. Flow cytometric analyses of H2B-GFP+ EryP revealed that Eklf heterozygosity results in the loss of Ter119 surface expression on EryP but not on EryD. Null mutation of Eklf resulted in abnormal expression of a range of surface proteins by EryP. In particular, several megakaryocyte markers were ectopically expressed by maturing Eklf-null EryP. Unexpectedly, the platelet tetraspanin CD9 was detected on nucleated wild-type EryP but not on mature EryD and thus provides a useful marker for purifying circulating EryP. We conclude that Eklf gene dosage is crucial for regulating the surface phenotype and molecular identity of maturing primitive erythroid cells.",
author = "Joan Isern and Fraser, {Stuart T.} and Zhiyong He and Hailan Zhang and Baron, {Margaret H.}",
year = "2010",
month = "11",
day = "11",
doi = "10.1182/blood-2010-04-281196",
language = "English (US)",
volume = "116",
pages = "3972--3980",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "19",

}

TY - JOUR

T1 - Dose-dependent regulation of primitive erythroid maturation and identity by the transcription factor Eklf

AU - Isern, Joan

AU - Fraser, Stuart T.

AU - He, Zhiyong

AU - Zhang, Hailan

AU - Baron, Margaret H.

PY - 2010/11/11

Y1 - 2010/11/11

N2 - The primitive erythroid (EryP) lineage is the first to differentiate during mammalian embryogenesis. Eklf/Klf1 is a transcriptional regulator that is essential for definitive erythropoiesis in the fetal liver. Dissection of the role(s) of Eklf within the EryP compartment has been confounded by the simultaneous presence of EryP and fetal liver-derived definitive erythroid (EryD) cells in the blood. To address this problem, we have distinguished EryP from their definitive counterparts by crossing Eklf+/- mutant and ε-globin:: histone H2B-GFP transgenic mice. Eklfdeficient EryP exhibit membrane ruffling and a failure to acquire the typical discoidal erythroid shape but they can enucleate. Flow cytometric analyses of H2B-GFP+ EryP revealed that Eklf heterozygosity results in the loss of Ter119 surface expression on EryP but not on EryD. Null mutation of Eklf resulted in abnormal expression of a range of surface proteins by EryP. In particular, several megakaryocyte markers were ectopically expressed by maturing Eklf-null EryP. Unexpectedly, the platelet tetraspanin CD9 was detected on nucleated wild-type EryP but not on mature EryD and thus provides a useful marker for purifying circulating EryP. We conclude that Eklf gene dosage is crucial for regulating the surface phenotype and molecular identity of maturing primitive erythroid cells.

AB - The primitive erythroid (EryP) lineage is the first to differentiate during mammalian embryogenesis. Eklf/Klf1 is a transcriptional regulator that is essential for definitive erythropoiesis in the fetal liver. Dissection of the role(s) of Eklf within the EryP compartment has been confounded by the simultaneous presence of EryP and fetal liver-derived definitive erythroid (EryD) cells in the blood. To address this problem, we have distinguished EryP from their definitive counterparts by crossing Eklf+/- mutant and ε-globin:: histone H2B-GFP transgenic mice. Eklfdeficient EryP exhibit membrane ruffling and a failure to acquire the typical discoidal erythroid shape but they can enucleate. Flow cytometric analyses of H2B-GFP+ EryP revealed that Eklf heterozygosity results in the loss of Ter119 surface expression on EryP but not on EryD. Null mutation of Eklf resulted in abnormal expression of a range of surface proteins by EryP. In particular, several megakaryocyte markers were ectopically expressed by maturing Eklf-null EryP. Unexpectedly, the platelet tetraspanin CD9 was detected on nucleated wild-type EryP but not on mature EryD and thus provides a useful marker for purifying circulating EryP. We conclude that Eklf gene dosage is crucial for regulating the surface phenotype and molecular identity of maturing primitive erythroid cells.

UR - http://www.scopus.com/inward/record.url?scp=78149446989&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78149446989&partnerID=8YFLogxK

U2 - 10.1182/blood-2010-04-281196

DO - 10.1182/blood-2010-04-281196

M3 - Article

C2 - 20720183

AN - SCOPUS:78149446989

VL - 116

SP - 3972

EP - 3980

JO - Blood

JF - Blood

SN - 0006-4971

IS - 19

ER -