Germ cells harvested from mouse embryonic genital ridges were mixed with disaggregated embryonic lung cells, and the reaggregates were cultured for 4-7 days. Germ cells derived from female embryos 10.5-13.5 days postcoitum (dpc) entered and progressed through meiotic prophase in vitro as in vivo, although with a 12- to 24-hr delay. If the cultures were maintained for 2-3 weeks, the germ cells developed into growing oocytes. When germ cells were taken from male embryos 10.5 and 11.5 dpc, they too entered and progressed through meiotic prophase, but germ cells from later embryos (12.5 and 13.5 dpc) developed as prospermatogonia, as in male genital ridges in vivo. When 11.5 dpc male genital ridges were disaggregated, reaggregated, and cultured in the same way as the lung reaggregates, the germ cells again entered meiotic prophase. We conclude that the male genital ridge at about 12 dpc produces a factor that inhibits entry of germ cells into meiosis, and that production of this factor is disrupted by prior disaggregation of the genital ridge. If a meiotic inducing substance is required for entry of germ cells into meiosis, it must be present in the male genital ridge as well as in the female genital ridge, and probably also in the lung.
In the early epiblast of female mice, one of the two X chromosomes is randomly inactivated by a Xist-dependent mechanism, involving the recruitment of Ezh2-Eed and the subsequent trimethylation of histone 3 on lysine 27 (H3K27me3). We demonstrate that this random inactivation process applies also to the primordial germ cell (PGC) precursors, located in the proximal region of the epiblast. PGC specification occurs at about embryonic day (E)7.5, in the extraembryonic mesoderm, after which the germ cells enter the endoderm of the invaginating hindgut. As they migrate towards the site of the future gonads, the XX PGCs gradually lose the H3K27me3 accumulation on the silent X chromosome. However, using a GFP transgene inserted into the X chromosome, we observed that the XX gonadal environment (independently of the gender) is important for the substantial reactivation of the inactive X chromosome between E11.5 and E13.5, but is not required for X-chromosome reactivation during the derivation of pluripotent embryonic germ cells. We describe in detail one of the key events during female PGC development, the epigenetic reprogramming of the X chromosome, and demonstrate the role of the XX somatic genital ridge in this process.
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