Characterization of expanded intermediate cell mass in zebrafish chordin morphant embryos

Anskar Y.H. Leung; Eric M. Mendenhall; Tommy T.F. Kwan; Raymond Liang; Craig E Eckfeldt; Eleanor Chen; Matthias Hammerschmidt; Suzanne Grindley; Stephen C. Ekker; Catherine M. Verfaillie

doi:10.1016/j.ydbio.2004.09.032

Characterization of expanded intermediate cell mass in zebrafish chordin morphant embryos

Anskar Y.H. Leung, Eric M. Mendenhall, Tommy T.F. Kwan, Raymond Liang, Craig E Eckfeldt, Eleanor Chen, Matthias Hammerschmidt, Suzanne Grindley, Stephen C. Ekker, Catherine M. Verfaillie

Medicine - Hematology, Oncology, Transplant

Research output: Contribution to journal › Article › peer-review

40 Scopus citations

Abstract

We investigated the mechanisms of intermediate cell mass (ICM) expansion in zebrafish chordin (Chd) morphant embryos and examined the role of BMPs in relation to this phenotype. At 24 h post-fertilization (hpf), the expanded ICM of embryos injected with chd morpholino (MO) (Chd^MO embryos) contained a monotonous population of hematopoietic progenitors. In situ hybridization showed that hematopoietic transcription factors were ubiquitously expressed in the ICM whereas vascular gene expression was confined to the periphery. BMP4 (but not BMP2b or 7) and smad5 mRNA were ectopically expressed in the Chd^MO ICM. At 48 hpf, monocytic cells were evident in both the ICM and circulation of Chd^MO but not WT embryos. While injection of BMP4 MO had no effect on WT hematopoiesis, co-injecting BMP4 with chd MOs significantly reduced ICM expansion. Microarray studies revealed a number of genes that were differentially expressed in Chd^MO and WT embryos and their roles in hematopoiesis has yet to be determined. In conclusion, the expanded ICM in Chd^MO embryos represented an expansion of embryonic hematopoiesis that was skewed towards a monocytic lineage. BMP4, but not BMP2b or 7, was involved in this process. The results provide ground for further research into the mechanisms of embryonic hematopoietic cell expansion.

Original language	English (US)
Pages (from-to)	235-254
Number of pages	20
Journal	Developmental Biology
Volume	277
Issue number	1
DOIs	https://doi.org/10.1016/j.ydbio.2004.09.032
State	Published - Jan 1 2005

Bibliographical note

Funding Information:
We thank Dr. Leonard Zon (Howard Hughes Medical Institute, Division of Hematology/Oncology) for the generous supply of plasmids for in situ hybridization and Prof. L.C. Chan, Dr. Edmond Ma and Mr. MB Chung (Department of Pathology, University of Hong Kong) for their technical support in tissue sectioning. We also thank Drs. Steve Farber and Chang-Gong Liu (Microarray Core Facility, Thomas Jefferson University) for performing the microarray and data analysis. This work was supported by NIH-PO1-CA-65493, the Tulloch Family, and The McKnight Foundation. AYH Leung was sponsored by the University of Hong Kong and the Croucher Foundation.

Keywords

BMP
Chordin
Differentiation
Gene expression
Hematopoiesis
Microarray
Morpholino
Zebrafish

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10.1016/j.ydbio.2004.09.032

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title = "Characterization of expanded intermediate cell mass in zebrafish chordin morphant embryos",

abstract = "We investigated the mechanisms of intermediate cell mass (ICM) expansion in zebrafish chordin (Chd) morphant embryos and examined the role of BMPs in relation to this phenotype. At 24 h post-fertilization (hpf), the expanded ICM of embryos injected with chd morpholino (MO) (ChdMO embryos) contained a monotonous population of hematopoietic progenitors. In situ hybridization showed that hematopoietic transcription factors were ubiquitously expressed in the ICM whereas vascular gene expression was confined to the periphery. BMP4 (but not BMP2b or 7) and smad5 mRNA were ectopically expressed in the ChdMO ICM. At 48 hpf, monocytic cells were evident in both the ICM and circulation of ChdMO but not WT embryos. While injection of BMP4 MO had no effect on WT hematopoiesis, co-injecting BMP4 with chd MOs significantly reduced ICM expansion. Microarray studies revealed a number of genes that were differentially expressed in ChdMO and WT embryos and their roles in hematopoiesis has yet to be determined. In conclusion, the expanded ICM in ChdMO embryos represented an expansion of embryonic hematopoiesis that was skewed towards a monocytic lineage. BMP4, but not BMP2b or 7, was involved in this process. The results provide ground for further research into the mechanisms of embryonic hematopoietic cell expansion.",

keywords = "BMP, Chordin, Differentiation, Gene expression, Hematopoiesis, Microarray, Morpholino, Zebrafish",

author = "Leung, {Anskar Y.H.} and Mendenhall, {Eric M.} and Kwan, {Tommy T.F.} and Raymond Liang and Eckfeldt, {Craig E} and Eleanor Chen and Matthias Hammerschmidt and Suzanne Grindley and Ekker, {Stephen C.} and Verfaillie, {Catherine M.}",

note = "Funding Information: We thank Dr. Leonard Zon (Howard Hughes Medical Institute, Division of Hematology/Oncology) for the generous supply of plasmids for in situ hybridization and Prof. L.C. Chan, Dr. Edmond Ma and Mr. MB Chung (Department of Pathology, University of Hong Kong) for their technical support in tissue sectioning. We also thank Drs. Steve Farber and Chang-Gong Liu (Microarray Core Facility, Thomas Jefferson University) for performing the microarray and data analysis. This work was supported by NIH-PO1-CA-65493, the Tulloch Family, and The McKnight Foundation. AYH Leung was sponsored by the University of Hong Kong and the Croucher Foundation. ",

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AU - Leung, Anskar Y.H.

AU - Mendenhall, Eric M.

AU - Kwan, Tommy T.F.

AU - Liang, Raymond

AU - Eckfeldt, Craig E

AU - Chen, Eleanor

AU - Hammerschmidt, Matthias

AU - Grindley, Suzanne

AU - Ekker, Stephen C.

AU - Verfaillie, Catherine M.

N1 - Funding Information: We thank Dr. Leonard Zon (Howard Hughes Medical Institute, Division of Hematology/Oncology) for the generous supply of plasmids for in situ hybridization and Prof. L.C. Chan, Dr. Edmond Ma and Mr. MB Chung (Department of Pathology, University of Hong Kong) for their technical support in tissue sectioning. We also thank Drs. Steve Farber and Chang-Gong Liu (Microarray Core Facility, Thomas Jefferson University) for performing the microarray and data analysis. This work was supported by NIH-PO1-CA-65493, the Tulloch Family, and The McKnight Foundation. AYH Leung was sponsored by the University of Hong Kong and the Croucher Foundation.

PY - 2005/1/1

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N2 - We investigated the mechanisms of intermediate cell mass (ICM) expansion in zebrafish chordin (Chd) morphant embryos and examined the role of BMPs in relation to this phenotype. At 24 h post-fertilization (hpf), the expanded ICM of embryos injected with chd morpholino (MO) (ChdMO embryos) contained a monotonous population of hematopoietic progenitors. In situ hybridization showed that hematopoietic transcription factors were ubiquitously expressed in the ICM whereas vascular gene expression was confined to the periphery. BMP4 (but not BMP2b or 7) and smad5 mRNA were ectopically expressed in the ChdMO ICM. At 48 hpf, monocytic cells were evident in both the ICM and circulation of ChdMO but not WT embryos. While injection of BMP4 MO had no effect on WT hematopoiesis, co-injecting BMP4 with chd MOs significantly reduced ICM expansion. Microarray studies revealed a number of genes that were differentially expressed in ChdMO and WT embryos and their roles in hematopoiesis has yet to be determined. In conclusion, the expanded ICM in ChdMO embryos represented an expansion of embryonic hematopoiesis that was skewed towards a monocytic lineage. BMP4, but not BMP2b or 7, was involved in this process. The results provide ground for further research into the mechanisms of embryonic hematopoietic cell expansion.

AB - We investigated the mechanisms of intermediate cell mass (ICM) expansion in zebrafish chordin (Chd) morphant embryos and examined the role of BMPs in relation to this phenotype. At 24 h post-fertilization (hpf), the expanded ICM of embryos injected with chd morpholino (MO) (ChdMO embryos) contained a monotonous population of hematopoietic progenitors. In situ hybridization showed that hematopoietic transcription factors were ubiquitously expressed in the ICM whereas vascular gene expression was confined to the periphery. BMP4 (but not BMP2b or 7) and smad5 mRNA were ectopically expressed in the ChdMO ICM. At 48 hpf, monocytic cells were evident in both the ICM and circulation of ChdMO but not WT embryos. While injection of BMP4 MO had no effect on WT hematopoiesis, co-injecting BMP4 with chd MOs significantly reduced ICM expansion. Microarray studies revealed a number of genes that were differentially expressed in ChdMO and WT embryos and their roles in hematopoiesis has yet to be determined. In conclusion, the expanded ICM in ChdMO embryos represented an expansion of embryonic hematopoiesis that was skewed towards a monocytic lineage. BMP4, but not BMP2b or 7, was involved in this process. The results provide ground for further research into the mechanisms of embryonic hematopoietic cell expansion.

KW - BMP

KW - Chordin

KW - Differentiation

KW - Gene expression

KW - Hematopoiesis

KW - Microarray

KW - Morpholino

KW - Zebrafish

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Characterization of expanded intermediate cell mass in zebrafish chordin morphant embryos

Abstract

Bibliographical note

Keywords

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