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Blimp1 is a critical determinant of the germ cell lineage in mice

Abstract

Germ cell fate in mice is induced in pluripotent epiblast cells in response to signals from extraembryonic tissues. The specification of approximately 40 founder primordial germ cells and their segregation from somatic neighbours are important events in early development. We have proposed that a critical event during this specification includes repression of a somatic programme that is adopted by neighbouring cells. Here we show that Blimp1 (also known as Prdm1), a known transcriptional repressor, has a critical role in the foundation of the mouse germ cell lineage, as its disruption causes a block early in the process of primordial germ cell formation. Blimp1-deficient mutant embryos form a tight cluster of about 20 primordial germ cell-like cells, which fail to show the characteristic migration, proliferation and consistent repression of homeobox genes that normally accompany specification of primordial germ cells. Furthermore, our genetic lineage-tracing experiments indicate that the Blimp1-positive cells originating from the proximal posterior epiblast cells are indeed the lineage-restricted primordial germ cell precursors.

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Figure 1: Expression of Blimp1 in gastrulating embryos.
Figure 2: Origin and allocation of Blimp1 –mEGFP cells.
Figure 3: Blimp1 -positive cells become TNAP- and stella-positive PGCs.
Figure 4: Blimp1 -positive cells are lineage-restricted PGCs.
Figure 5: Loss of PGCs in Blimp1 deficient embryos.
Figure 6: Aberrant phenotype of Blimp1 deficient cells.

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Acknowledgements

We thank S. Chuva de Sousa Lopes, K. Nakao, H. Miyachi and R. Nakayama for technical help, and T. Nakano for anti-stella/PGC7 antibody. B.P. was supported by a Wellcome Trust PhD studentship. M.A.S. is funded by the BBSRC, the Wellcome Trust and the EU Epigenome Programme. M.S. is supported by the Ministry of Education, Culture, Sports, Science and Technology, and a PRESTO grant by the JST. D.O'C. acknowledges the support of the Irvington Institute for Immunological Research and is their National Genetics Foundation Fellow. Thanks to K. Lawson and A. McLaren for discussions and critical comments.

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Correspondence to Mitinori Saitou or M. Azim Surani.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Methods

This file contains additional information on the methods used in this study, including the antibodies used and the generation of transgenic and knockout mouse strains described in the paper. (DOC 33 kb)

Supplementary Figure Legends

Legends to accompany the below Supplementary Figures. (DOC 35 kb)

Supplementary Figure S1

Single cell analysis of germ cell marker expression during PGC-specification and fragilis in situ hybridisations. (JPG 64 kb)

Supplementary Figure S2

Description of the Blimp-1-mEGFP transgene and its expression during PGC-specification. (JPG 57 kb)

Supplementary Figure S3

Co-staining of TNAP and Blimp-1-mEGFP in MB stage embryos. (JPG 64 kb)

Supplementary Figure S4

Creation of Blimp-1 mutant allele. The figure contains a description of the targeting strategy and verification of successful targeting by Southern blotting and RT–PCR. (JPG 67 kb)

Supplementary Figure S5

Comparison of E9.5 embryos generated from Blimp-1 control (loxP/loxP) and null (-/-) ES-cells injected into tetraploid blastocysts. TNAP-stainings revealled a lack or gross reduction of PGCs in the mutants. (JPG 52 kb)

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Ohinata, Y., Payer, B., O'Carroll, D. et al. Blimp1 is a critical determinant of the germ cell lineage in mice. Nature 436, 207–213 (2005). https://doi.org/10.1038/nature03813

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