TY - JOUR
T1 - Esrrb function is required for proper primordial germ cell development in presomite stage mouse embryos
AU - Okamura, Eiichi
AU - Tam, Oliver H.
AU - Posfai, Eszter
AU - Li, Lingyu
AU - Cockburn, Katie
AU - Lee, Cheryl Q.E.
AU - Garner, Jodi
AU - Rossant, Janet
N1 - Funding Information:
This work was supported by JSPS KAKENHI Grant Number 15H06447 (E.O.). E.O. was supported by a postdoctoral fellowship from the Uehara Memorial Foundation and Study Abroad Grant Program from BioLegend/Tomy Digital Biology. O.H.T was supported by a fellowship of the Human Frontier Science Program. We thank the Transgenic Core lab headed by Marina Gertsenstein at The Centre for Phenogenomics (TCP), Toronto for transgenic services, The Centre for Applied Genomics (TCAG) facility at the Hospital for Sick Children and Support Centre for Advanced Medical Sciences of Tokushima University Graduate School of Biomedical Sciences for technical support. We thank all members of the Department of Genetic Engineering and Animal Research Resources, Tokushima University for help and valuable discussions.
Funding Information:
This work was supported by JSPS KAKENHI Grant Number 15H06447 (E.O.). E.O. was supported by a postdoctoral fellowship from the Uehara Memorial Foundation and Study Abroad Grant Program from BioLegend/Tomy Digital Biology . O.H.T was supported by a fellowship of the Human Frontier Science Program . We thank the Transgenic Core lab headed by Marina Gertsenstein at The Centre for Phenogenomics (TCP), Toronto for transgenic services, The Centre for Applied Genomics (TCAG) facility at the Hospital for Sick Children and Support Centre for Advanced Medical Sciences of Tokushima University Graduate School of Biomedical Sciences for technical support. We thank all members of the Department of Genetic Engineering and Animal Research Resources, Tokushima University for help and valuable discussions.
Publisher Copyright:
© 2019 Elsevier Inc.
PY - 2019/11/15
Y1 - 2019/11/15
N2 - Estrogen related receptor beta (Esrrb) is an orphan nuclear receptor that is required for self-renewal and pluripotency in mouse embryonic stem (ES) cells. However, in the early post-implantation mouse embryo, Esrrb is specifically expressed in the extraembryonic ectoderm (ExE) and plays a crucial role in trophoblast development. Previous studies showed that Esrrb is also required to maintain trophoblast stem (TS) cells, the in vitro stem cell model of the early trophoblast lineage. In order to identify regulatory targets of Esrrb in vivo, we performed microarray analysis of Esrrb-null versus wild-type post-implantation ExE, and identified 30 genes down-regulated in Esrrb-mutants. Among them is Bmp4, which is produced by the ExE and known to be critical for primordial germ cell (PGC) specification in vivo. We further identified an enhancer region bound by Esrrb at the Bmp4 locus by performing Esrrb ChIP-seq and luciferase reporter assay using TS cells. Finally, we established a knockout mouse line in which the enhancer region was deleted using CRISPR/Cas9 technology. Both Esrrb-null embryos and enhancer knockout embryos expressed lower levels of Bmp4 in the ExE, and had reduced numbers of PGCs. These results suggested that Esrrb functions as an upstream factor of Bmp4 in the ExE, regulating proper PGC development in mice.
AB - Estrogen related receptor beta (Esrrb) is an orphan nuclear receptor that is required for self-renewal and pluripotency in mouse embryonic stem (ES) cells. However, in the early post-implantation mouse embryo, Esrrb is specifically expressed in the extraembryonic ectoderm (ExE) and plays a crucial role in trophoblast development. Previous studies showed that Esrrb is also required to maintain trophoblast stem (TS) cells, the in vitro stem cell model of the early trophoblast lineage. In order to identify regulatory targets of Esrrb in vivo, we performed microarray analysis of Esrrb-null versus wild-type post-implantation ExE, and identified 30 genes down-regulated in Esrrb-mutants. Among them is Bmp4, which is produced by the ExE and known to be critical for primordial germ cell (PGC) specification in vivo. We further identified an enhancer region bound by Esrrb at the Bmp4 locus by performing Esrrb ChIP-seq and luciferase reporter assay using TS cells. Finally, we established a knockout mouse line in which the enhancer region was deleted using CRISPR/Cas9 technology. Both Esrrb-null embryos and enhancer knockout embryos expressed lower levels of Bmp4 in the ExE, and had reduced numbers of PGCs. These results suggested that Esrrb functions as an upstream factor of Bmp4 in the ExE, regulating proper PGC development in mice.
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U2 - 10.1016/j.ydbio.2019.07.008
DO - 10.1016/j.ydbio.2019.07.008
M3 - Article
C2 - 31315026
AN - SCOPUS:85069870567
SN - 0012-1606
VL - 455
SP - 382
EP - 392
JO - Developmental Biology
JF - Developmental Biology
IS - 2
ER -