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No imprinted XIST expression in pigs: biallelic XIST expression in early embryos and random X inactivation in placentas

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Abstract

Dosage compensation, which is achieved by X-chromosome inactivation (XCI) in female mammals, ensures balanced X-linked gene expression levels between the sexes. Although eutherian mammals commonly display random XCI in embryonic and adult tissues, imprinted XCI has also been identified in extraembryonic tissues of mouse, rat, and cow. Little is known about XCI in pigs. Here, we sequenced the porcine XIST gene and identified an insertion/deletion mutation between Asian- and Western-origin pig breeds. Allele-specific analysis revealed biallelic XIST expression in porcine ICSI blastocysts. To investigate the XCI pattern in porcine placentas, we performed allele-specific RNA sequencing analysis on individuals from reciprocal crosses between Duroc and Rongchang pigs. Our results were the first to reveal that random XCI occurs in the placentas of pigs. Next, we investigated the H3K27me3 histone pattern in porcine blastocysts, showing that only 17–31.8% cells have attained XCI. The hypomethylation status of an important XIST DMR (differentially methylated region) in gametes and early embryos demonstrated that no methylation is pre-deposited on XIST in pigs. Our findings reveal that the XCI regulation mechanism in pigs is different from that in mice and highlight the importance of further study of the mechanisms regulating XCI during early porcine embryo development.

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Abbreviations

DMR:

Differentially methylated region

E:

Embryonic day

H3K27me3:

Histone H3 lysine 27 trimethylation

ICSI:

Intracytoplasmic sperm injection

ICR:

Imprinting control region

IVF:

In vitro fertilization

Xic:

X inactivation center

PA:

Parthenogenetic

PRC2:

Polycomb repressive complex-2

RFLP:

Restriction fragment length polymorphism

SNP:

Single nucleotide polymorphism

TSS:

Transcription start site

TTS:

Transcription termination site

Xa:

Active X chromosome

XCI:

X-chromosome inactivation

Xi:

Inactive X chromosome

XIST :

X-inactive specific transcript

Xm:

Maternal X chromosome

Xp:

Paternal X chromosome

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Acknowledgements

This work was supported by The Agricultural Science and Technology Innovation Program (ASTIP-IAS06, CAAS-XTCX2016010-02) and Transgenic Research Grant 2016ZX08010001.

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Author notes

  1. Huiying Zou, Dawei Yu, Xuguang Du, and Jing Wang have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Huiying Zou, Yangyang Wang, Huitao Xu, Shanjiang Zhao, Yunwei Pang, Yan Liu, Haisheng Hao, Xueming Zhao, Weihua Du & Huabin Zhu

  2. State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China

    Huiying Zou, Xuguang Du, Jing Wang, Yunping Dai, Ning Li & Sen Wu

  3. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China

    Dawei Yu & Yunxuan Zhao

  4. Chongqing Academy of Animal Science, Chongqing, 402460, China

    Lei Chen

  5. Beijing Advanced Innovation Center for Food Nutrition and Human Health, China Agricultural University, Beijing, 100193, China

    Sen Wu

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  1. Huiying Zou
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Contributions

HBZ, SW, and NL designed the research; HYZ, DWY, XGD, JW, LC, YYW, HTX, YXZ, SJZ, YWP, YL, HSH, XMZ, WHD, and YPD performed the experiments and analyzed data. HBZ, SW, HYZ, DWY, XGD, and JW wrote the paper. All authors read and approved the final manuscript.

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Correspondence to Sen Wu or Huabin Zhu.

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Zou, H., Yu, D., Du, X. et al. No imprinted XIST expression in pigs: biallelic XIST expression in early embryos and random X inactivation in placentas. Cell. Mol. Life Sci. 76, 4525–4538 (2019). https://doi.org/10.1007/s00018-019-03123-3

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