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. 2007 Jul;3(7):e116.
doi: 10.1371/journal.pgen.0030116.

X chromosome reactivation initiates in nascent primordial germ cells in mice

Michihiko Sugimoto  1 , Kuniya Abe
Affiliations

X chromosome reactivation initiates in nascent primordial germ cells in mice

Michihiko Sugimoto et al. PLoS Genet. 2007 Jul.

Abstract

During primordial germ cell (PGC) development, epigenetic reprogramming events represented by X chromosome reactivation and erasure of genomic imprinting are known to occur. Although precise timing is not given, X reactivation is thought to take place over a short period of time just before initiation of meiosis. Here, we show that the cessation of Xist expression commences in nascent PGCs, and re-expression of some X-linked genes begins in newly formed PGCs VSports手机版. The X reactivation process was not complete in E14. 5 PGCs, indicating that X reactivation in developing PGCs occurs over a prolonged period. These results set the reactivation timing much earlier than previously thought and suggest that X reactivation may involve slow passive steps. .

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Conflict of interest statement (VSports手机版)

Competing interests V体育安卓版. The authors have declared that no competing interests exist.

Figures (V体育官网入口)

Figure 1
Figure 1. Results of Whole-Mount RNA FISH with Xist Probe
(A–E) An E5.5 embryo-expressing methyl-CpG binding domain (MBD)-GFP fusion protein (green) under the control of Oct4 promoter is shown (S. Kobayakawa and K. Abe, unpublished data). This transgene was used to visualize the nucleus of epiblast. (A) Shown is a single optical section merged with DIC image. Images in panels (B–D) were obtained by projecting ten optical slices at different depths of the specimen shown in (A). (E) A projection image from all 230 optical slices is presented. Red is Xist RNA signal, green is GFP fluorescence. (F–J) Whole-mount RNA FISH used Xist (red) and Cot-1 (white) probes combined with immunofluorescence for PGC markers (green) with antibodies against Oct4 (F, G), Stella (H), and RFP (I). Images in each panel were obtained by projecting ten optical slices. Xist RNA did not accumulate in the nuclei of E10.5 PGCs (F). Xist (+) PGCs (arrow), Xist (±) PGCs (arrowhead), and also Xist (−) cells, were present at E8.5 (G), E7.75 (H), and E7.0 (I). (J) Percentages of Xist (+), Xist (±), and Xist (−) cells in the cell populations tested. As controls, the percentages of cells with each Xist signal pattern are indicated for the embryonic ectoderm (em. ec.) and extraembryonic ectoderm (ex. ec.) in E7.75 embryos and for the somatic cells surrounding the PGCs in E10.5 embryos. Cells with scattered Xist signals in their nuclei are classified as “others.”
Figure 2
Figure 2. Biallelic Expression of X-Linked Genes in PGCs Revealed by Single-Cell RT-PCR
Some X-linked genes are already expressed biallelically (asterisks) in some PGCs at E7.75, but that not all X-linked genes are biallelically expressed, even at E14.5. A total of four PGCs at each stage are indicated. Biallelic expression is marked with asterisks. The relative positions of the X-linked genes are roughly indicated.
Figure 3
Figure 3. Expression Patterns in PGCs at Each Stage
(A) Ratio of cells biallelically expressing X-linked genes is presented. (B) A schematic shows the transition of gene expression patterns during PGC development. Each circle graph indicates the ratio of cells that are positive (yellow) and negative (black) for each gene, and biallelically (red) and monoallelically (blue) expressed in cells positive for each gene.
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References

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