Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.

In mouse female preimplantation embryos, the paternal X chromosome (Xp) is silenced by imprinted X chromosome inactivation (iXCI). This requires production of the noncoding Xist RNA in cis, from the Xp. The Xist locus on the maternally inherited X chromosome (Xm) is refractory to activation due to t...

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Main Authors: Federica Federici, Aristea Magaraki, Evelyne Wassenaar, Catherina J H van Veen-Buurman, Christine van de Werken, Esther B Baart, Joop S E Laven, J Anton Grootegoed, Joost Gribnau, Willy M Baarends
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2016-10-01
Series:PLoS Genetics
Online Access:https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006358&type=printable
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author Federica Federici
Aristea Magaraki
Evelyne Wassenaar
Catherina J H van Veen-Buurman
Christine van de Werken
Esther B Baart
Joop S E Laven
J Anton Grootegoed
Joost Gribnau
Willy M Baarends
author_facet Federica Federici
Aristea Magaraki
Evelyne Wassenaar
Catherina J H van Veen-Buurman
Christine van de Werken
Esther B Baart
Joop S E Laven
J Anton Grootegoed
Joost Gribnau
Willy M Baarends
author_sort Federica Federici
collection DOAJ
description In mouse female preimplantation embryos, the paternal X chromosome (Xp) is silenced by imprinted X chromosome inactivation (iXCI). This requires production of the noncoding Xist RNA in cis, from the Xp. The Xist locus on the maternally inherited X chromosome (Xm) is refractory to activation due to the presence of an imprint. Paternal inheritance of an Xist deletion (XpΔXist) is embryonic lethal to female embryos, due to iXCI abolishment. Here, we circumvented the histone-to-protamine and protamine-to-histone transitions of the paternal genome, by fertilization of oocytes via injection of round spermatids (ROSI). This did not affect initiation of XCI in wild type female embryos. Surprisingly, ROSI using ΔXist round spermatids allowed survival of female embryos. This was accompanied by activation of the intact maternal Xist gene, initiated with delayed kinetics, around the morula stage, resulting in Xm silencing. Maternal Xist gene activation was not observed in ROSI-derived males. In addition, no Xist expression was detected in male and female morulas that developed from oocytes fertilized with mature ΔXist sperm. Finally, the expression of the X-encoded XCI-activator RNF12 was enhanced in both male (wild type) and female (wild type as well as XpΔXist) ROSI derived embryos, compared to in vivo fertilized embryos. Thus, high RNF12 levels may contribute to the specific activation of maternal Xist in XpΔXist female ROSI embryos, but upregulation of additional Xp derived factors and/or the specific epigenetic constitution of the round spermatid-derived Xp are expected to be more critical. These results illustrate the profound impact of a dysregulated paternal epigenome on embryo development, and we propose that mouse ROSI can be used as a model to study the effects of intergenerational inheritance of epigenetic marks.
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spelling doaj-art-9d08fb3963e04fdaa5e1f586a9987f9d2025-08-20T03:04:53ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042016-10-011210e100635810.1371/journal.pgen.1006358Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.Federica FedericiAristea MagarakiEvelyne WassenaarCatherina J H van Veen-BuurmanChristine van de WerkenEsther B BaartJoop S E LavenJ Anton GrootegoedJoost GribnauWilly M BaarendsIn mouse female preimplantation embryos, the paternal X chromosome (Xp) is silenced by imprinted X chromosome inactivation (iXCI). This requires production of the noncoding Xist RNA in cis, from the Xp. The Xist locus on the maternally inherited X chromosome (Xm) is refractory to activation due to the presence of an imprint. Paternal inheritance of an Xist deletion (XpΔXist) is embryonic lethal to female embryos, due to iXCI abolishment. Here, we circumvented the histone-to-protamine and protamine-to-histone transitions of the paternal genome, by fertilization of oocytes via injection of round spermatids (ROSI). This did not affect initiation of XCI in wild type female embryos. Surprisingly, ROSI using ΔXist round spermatids allowed survival of female embryos. This was accompanied by activation of the intact maternal Xist gene, initiated with delayed kinetics, around the morula stage, resulting in Xm silencing. Maternal Xist gene activation was not observed in ROSI-derived males. In addition, no Xist expression was detected in male and female morulas that developed from oocytes fertilized with mature ΔXist sperm. Finally, the expression of the X-encoded XCI-activator RNF12 was enhanced in both male (wild type) and female (wild type as well as XpΔXist) ROSI derived embryos, compared to in vivo fertilized embryos. Thus, high RNF12 levels may contribute to the specific activation of maternal Xist in XpΔXist female ROSI embryos, but upregulation of additional Xp derived factors and/or the specific epigenetic constitution of the round spermatid-derived Xp are expected to be more critical. These results illustrate the profound impact of a dysregulated paternal epigenome on embryo development, and we propose that mouse ROSI can be used as a model to study the effects of intergenerational inheritance of epigenetic marks.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006358&type=printable
spellingShingle Federica Federici
Aristea Magaraki
Evelyne Wassenaar
Catherina J H van Veen-Buurman
Christine van de Werken
Esther B Baart
Joop S E Laven
J Anton Grootegoed
Joost Gribnau
Willy M Baarends
Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
PLoS Genetics
title Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
title_full Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
title_fullStr Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
title_full_unstemmed Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
title_short Round Spermatid Injection Rescues Female Lethality of a Paternally Inherited Xist Deletion in Mouse.
title_sort round spermatid injection rescues female lethality of a paternally inherited xist deletion in mouse
url https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006358&type=printable
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