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Inhibition of LINE-1 retrotransposition represses telomere reprogramming during mouse 2-cell embryo development

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Abstract

Purpose

To investigate whether inhibition of LINE-1 affects telomere reprogramming during 2-cell embryo development.

Methods

Mouse zygotes were cultured with or without 1 µM azidothymidine (AZT) for up to 15 h (early 2-cell, G1/S) or 24 h (late 2-cell, S/G2). Gene expression and DNA copy number were determined by RT-qPCR and qPCR respectively. Immunostaining and telomeric PNA-FISH were performed for co-localization between telomeres and ZSCAN4 or LINE-1-Orf1p.

Results

LINE-1 copy number was remarkably reduced in later 2-cell embryos by exposure to 1 µM AZT, and telomere lengths in late 2-cell embryos with AZT were significantly shorter compared to control embryos (P = 0.0002). Additionally, in the absence of LINE-1 inhibition, Dux, Zscan4, and LINE-1 were highly transcribed in early 2-cell embryos, as compared to late 2-cell embryos (P < 0.0001), suggesting that these 2-cell genes are activated at the early 2-cell stage. However, in early 2-cell embryos with AZT treatment, mRNA levels of Dux, Zscan4, and LINE-1 were significantly decreased. Furthermore, both Zscan4 and LINE-1 encoded proteins localized to telomere regions in 2-cell embryos, but this co-localization was dramatically reduced after AZT treatment (P < 0.001).

Conclusions

Upon inhibition of LINE-1 retrotransposition in mouse 2-cell embryos, Dux, Zscan4, and LINE-1 were significantly downregulated, and telomere elongation was blocked. ZSCAN4 foci and their co-localization with telomeres were also significantly decreased, indicating that ZSCAN4 is an essential component of the telomere reprogramming that occurs in mice at the 2-cell stage. Our findings also suggest that LINE-1 may directly contribute to telomere reprogramming in addition to regulating gene expression.

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Funding

This study was supported by March of Dimes Grant # 6-FY14-432 and the Stanley H. Kaplan Endowment.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, New York University Grossman School of Medicine, New York, NY, 10016, USA

    Fang Wang, Isaac J. Chamani, Danxia Luo, Kasey Chan & David L. Keefe

  2. Human Reproduction Division, Department of Gynecology and Obstetrics, Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, Ribeirão Preto, Brazil

    Paula Andrea Navarro

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  1. Fang Wang
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Wang, F., Chamani, I.J., Luo, D. et al. Inhibition of LINE-1 retrotransposition represses telomere reprogramming during mouse 2-cell embryo development. J Assist Reprod Genet 38, 3145–3153 (2021). https://doi.org/10.1007/s10815-021-02331-w

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