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Ewing sarcoma protein ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo.

Azuma M, Embree LJ, Sabaawy H, Hickstein DD - PLoS ONE (2007)

Bottom Line: We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development.The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human.Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development.

View Article: PubMed Central - PubMed

Affiliation: Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT

Background: The Ewing sarcoma breakpoint region 1 gene (EWSR1), also known as EWS, is fused to a number of different partner genes as a result of chromosomal translocation in diverse sarcomas. Despite the involvement of EWSR1 in these diverse sarcomas, the in vivo function of wild type EWSR1 remains unclear.

Principal findings: We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development. Morpholino induced knockdown of both zebrafish ewsr1 genes led to mitotic defects with multipolar or otherwise abnormal mitotic spindles starting from the bud stage (10 hour post-fertilization (hpf)). The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human.

Conclusions: Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development.

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Related in: MedlinePlus

Initial Maternal and Subsequent Ubiquitious Expression of ewsr1a and ewsr1b mRNAs in Zebrafish Embryonic Development.The images are lateral views of embryos stained by in situ hybridization using anti-sense RNA (a–e, k-o) or sense RNA (f–j, p–t) for ewsr1a and ewsr1b. The ewsr1a and ewsr1b were expressed ubiquitously at 4 cell (a and k), 256 cell (b and l), shield (c and m), 5 somite stage (d and s) and 24hpf (e and t). asRNA: anti-sense RNA, sRNA: sense RNA
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pone-0000979-g002: Initial Maternal and Subsequent Ubiquitious Expression of ewsr1a and ewsr1b mRNAs in Zebrafish Embryonic Development.The images are lateral views of embryos stained by in situ hybridization using anti-sense RNA (a–e, k-o) or sense RNA (f–j, p–t) for ewsr1a and ewsr1b. The ewsr1a and ewsr1b were expressed ubiquitously at 4 cell (a and k), 256 cell (b and l), shield (c and m), 5 somite stage (d and s) and 24hpf (e and t). asRNA: anti-sense RNA, sRNA: sense RNA

Mentions: Human EWSR1 mRNA expression is ubiquitous in adult tissues [8]. However, there is no description of EWSR1 mRNA expression during embryonic development. We determined the spatial and temporal expression patterns of zebrafish ewsr1a and ewsr1b during development using whole-mount in situ hybridization. Both ewsr1a and ewsr1b were maternally expressed as early as the one cell stage (data not shown), and both genes continue to be expressed ubiquitously throughout early embryonic development (Figure 2, a∼e and k∼o).


Ewing sarcoma protein ewsr1 maintains mitotic integrity and proneural cell survival in the zebrafish embryo.

Azuma M, Embree LJ, Sabaawy H, Hickstein DD - PLoS ONE (2007)

Initial Maternal and Subsequent Ubiquitious Expression of ewsr1a and ewsr1b mRNAs in Zebrafish Embryonic Development.The images are lateral views of embryos stained by in situ hybridization using anti-sense RNA (a–e, k-o) or sense RNA (f–j, p–t) for ewsr1a and ewsr1b. The ewsr1a and ewsr1b were expressed ubiquitously at 4 cell (a and k), 256 cell (b and l), shield (c and m), 5 somite stage (d and s) and 24hpf (e and t). asRNA: anti-sense RNA, sRNA: sense RNA
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1991596&req=5

pone-0000979-g002: Initial Maternal and Subsequent Ubiquitious Expression of ewsr1a and ewsr1b mRNAs in Zebrafish Embryonic Development.The images are lateral views of embryos stained by in situ hybridization using anti-sense RNA (a–e, k-o) or sense RNA (f–j, p–t) for ewsr1a and ewsr1b. The ewsr1a and ewsr1b were expressed ubiquitously at 4 cell (a and k), 256 cell (b and l), shield (c and m), 5 somite stage (d and s) and 24hpf (e and t). asRNA: anti-sense RNA, sRNA: sense RNA
Mentions: Human EWSR1 mRNA expression is ubiquitous in adult tissues [8]. However, there is no description of EWSR1 mRNA expression during embryonic development. We determined the spatial and temporal expression patterns of zebrafish ewsr1a and ewsr1b during development using whole-mount in situ hybridization. Both ewsr1a and ewsr1b were maternally expressed as early as the one cell stage (data not shown), and both genes continue to be expressed ubiquitously throughout early embryonic development (Figure 2, a∼e and k∼o).

Bottom Line: We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development.The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human.Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development.

View Article: PubMed Central - PubMed

Affiliation: Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT

Background: The Ewing sarcoma breakpoint region 1 gene (EWSR1), also known as EWS, is fused to a number of different partner genes as a result of chromosomal translocation in diverse sarcomas. Despite the involvement of EWSR1 in these diverse sarcomas, the in vivo function of wild type EWSR1 remains unclear.

Principal findings: We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development. Morpholino induced knockdown of both zebrafish ewsr1 genes led to mitotic defects with multipolar or otherwise abnormal mitotic spindles starting from the bud stage (10 hour post-fertilization (hpf)). The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human.

Conclusions: Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development.

Show MeSH
Related in: MedlinePlus