Abstract

During early Xenopus laevis embryogenesis both nuclear and cell volumes decrease with the nuclear-to-cytoplasmic (N/C) volume ratio reaching a maximum at the midblastula transition (MBT). At the MBT, embryonic transcription is upregulated and cell cycles lengthen. Early studies demonstrated a role for the DNA-to-cytoplasmic ratio in the control of MBT timing. By altering nuclear size, we previously showed that the N/C volume ratio also contributes to proper MBT timing. Here we examine the relative contributions of nuclear size and DNA amount to MBT timing by simultaneously altering nuclear size and ploidy in X. laevis embryos. Compared to diploid embryos, haploids exhibited a delay in both zygotic gene expression and cell cycle lengthening, while diploid embryos with increased N/C volume ratios showed early expression of zygotic genes and premature lengthening of cell cycles. Interestingly, haploids with increased N/C volume ratios exhibited an intermediate effect on the timing of zygotic gene expression and cell cycle lengthening. Decreasing nuclear size in post-MBT haploid embryos caused a further delay in cell cycle lengthening and the expression of some zygotic genes. Our data suggest that both the N/C volume ratio and DNA amount contribute to the regulation of MBT timing with neither parameter being dominant.

Highlights

  • From stage 4 to stage 8, nuclear and cytoplasmic volumes decrease by 3-fold and ~70-fold, respectively

  • These results suggest that both DNA amount and the N/C volume ratio contribute to proper midblastula transition (MBT) timing, with neither parameter having a dominant effect

  • One-cell X. laevis haploid and diploid embryos generated from the same batch of eggs were microinjected with different mRNAs expressing nuclear scaling factors to increase or decrease nuclear size

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Summary

Transition Timing in Xenopus laevis

During early Xenopus laevis embryogenesis both nuclear and cell volumes decrease with the nuclear-tocytoplasmic (N/C) volume ratio reaching a maximum at the midblastula transition (MBT). Haploids with increased N/C volume ratios exhibited an intermediate effect on the timing of zygotic gene expression and cell cycle lengthening. We previously altered nuclear size in early X. laevis embryos and showed that this affected the timing of the onset of zygotic transcription and cell cycle lengthening[6], implicating the N/C volume ratio in the control of MBT timing. To test the relative contributions of DNA amount and the N/C volume ratio to MBT timing, we simultaneously altered nuclear size and ploidy in early X. laevis embryos and examined the timing of zygotic transcription by in situ hybridization and qRT-PCR and the onset of longer cell cycles by bright-field time-lapse microscopy. Zygotic gene expression and cell cycle lengthening were delayed in haploid embryos and occurred prematurely in embryos with increased N/C volume ratios. All nuclear size differences between haploids were statistically significant by p < 0.001

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