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A comprehensive, non-invasive visualization of primordial germ cell development in mice by the Blimp1-mVenus and stella-ECFP double transgenic reporter

Y Ohinata, Laboratory for Mammalian Germ Cell Biology, Center for Developmental Biology, RIKEN Kobe Institute, Kobe, Japan
M Sano, Laboratory for Mammalian Germ Cell Biology, Center for Developmental Biology, RIKEN Kobe Institute, Kobe, Japan
M Shigeta, Laboratory for Mammalian Germ Cell Biology, Center for Developmental Biology, RIKEN Kobe Institute, Kobe, Japan
K Yamanaka, Laboratory for Mammalian Germ Cell Biology, Center for Developmental Biology, RIKEN Kobe Institute, Kobe, Japan
M Saitou, Laboratory for Mammalian Germ Cell Biology, Center for Developmental Biology, RIKEN Kobe Institute , Kobe, 650-0047, Japan

Correspondence: Mitinori Saitou, Email: saitou{at}cdb.riken.jp

Abstract

The ability to monitor the development of a given cell lineage in a non-invasive manner by fluorescent markers both in vivo and in vitro provides a great advantage for the analysis of the lineage of interest. To date, a number of transgenic or knock-in mouse strains in which developing germ cells are marked with fluorescent reporters have been generated. We here describe a novel double transgenic reporter mouse strain that expresses membrane-targeted Venus (mVenus), a brighter variant of YFP, under the control of Blimp1 regulatory elements and ECFP under the control of stella. The double transgenic strain unambiguously marked Blimp1 expression in the lineage-restricted precursors of primordial germ cells (PGCs) in the proximal epiblast at embryonic day (E) 6.25 and specifically illuminated Blimp1- and stella-positive migrating PGCs after E8.5. The double transgenic reporter also precisely recapitulated dynamic embryonic expression of Blimp1 outside the germ cell lineage. Moreover, we derived ES cells that bore both transgenes. These cells made a robust contribution both to the germ and somatic cell lineages in chimeras with accurate Blimp1-mVenus and stella-ECFP expression. The transgenic strain and the ES cells will serve as valuable experimental materials not only for analyzing the origin and properties of the germ cell lineage in vivo but also for establishing a culture system to efficiently induce proper germ cells with temporally coordinated Blimp1 and stella expression in vitro.