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- W2976572087 abstract "During preimplantation embryo development, the telomere aging clock resets and 2-cell genes establish totipotency. Like most long -lived cells, oocytes have short telomeres, but telomeres elongate markedly during early embryo development, even in the absence of telomerase (1). Recent studies report that the retrotransposon, LINE-1, is activated after fertilization, when telomeres elongate. Retrotransposons protect chromosome ends in many species, so we hypothesized that inhibiting retrotransposition of LINE-1 during mouse early embryos would disrupt telomere reprogramming in mouse preimplantation embryos. We also evaluated the effect of LINE-1 on the 2-cell, totipotency genes. Laboratory experiment Sixty thawed mouse zygotes (B6C3F1 X B6D2F1) (Embryotech Laboratories) were grouped (15 zygotes/group) and cultured in Global medium containing 0.4% BSA with or without 1μM of the reverse transcriptase inhibitor, AZT, for up to 15 (mid 2-cell stage) or 24 hours (late 2-cell stage). DNA and mRNA were isolated from embryos. Gene expression was measured by RT-qPCR, and LINE-1 copy number and telomere length were measured by qPCR. Data (Mean±SEM) was analized with GraphPad Prism 8 software by one-way ANOVA Tukey's multiple comparisons test. As expected, AZT inhibited LINE-1 copy number in late 2-cell embryos compared to controls (1.000±0.113 vs 1.942±0.089, P<0.0001). Telomeres were longer in late 2-cell (1.563±0.107) than mid 2-cell embryos (1.128±0.059) (P=0.001), consistent with prior observations of progressive telomere elongation during early development(1). AZT blocked telomere elongation between mid- and late-2 cell stages stage (1.152±0.039 vs 1.000±0.107, P=0.1994). Telomere length of late 2-cell embryos exposed to AZT was shorter than untreated controls (1.000±0.107 vs 1.563±0.107, P=0.0002). Transcripts of LINE-1, as well as the 2-cell genes, DUX and Zscan4, were highly expressed in mid- 2-cell compared to late 2-cell embryos (P <0.0001), regardless of inhibition of LINE-1. This suggests that 2-cell genes are activated at synthesis phase of the 2-cell stage. Moreover, AZT decreased expression of DUX, Zscan4d and LINE-1 in mid- 2-cell embryos (0.312±0.021, 0.727±0.055 and 1.266±0.066 respectively) compared to control embryos (1.913±0.197, 1.912±0.202 and 1.913+0.216 respectively, P<0.001). The retrotransposon, LINE-1, regulates telomere lengthening during preimplantation embryo development- inhibition of LINE-1 synthesis by AZT blocks telomere elongation. AZT also inhibits the 2-cell genes, DUX and Zscan4d, suggesting that LINE-1 is essential not only for telomere reprogramming but also for the establishment of totipotency during early development." @default.
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- W2976572087 date "2019-09-01" @default.
- W2976572087 modified "2023-10-18" @default.
- W2976572087 title "Inhibition of line-1 transposition blocks telomere elongation and downregulates totipotency genes during mouse embryo development" @default.
- W2976572087 doi "https://doi.org/10.1016/j.fertnstert.2019.07.448" @default.
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