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Updated: Feb 6, 2026

Xenopus laevis Egg Extract Preparation and Live Imaging Methods for Visualizing Dynamic Cytoplasmic Organization
Published on: June 6, 2021
Rapid pronucleus assembly using cytoplasmic RNAs in fertilized eggs of Xenopus laevis
Mizuki Ikeda1, Yuto Tanaka1, Tatsuya Shohoji1
1Evolutionary Cell Biology Laboratory, Faculty of Science, Yamaguchi University, Yoshida 1677-1, Yamaguchi city, 753-8512, Japan.
Abstract:
The small hypercondensed sperm nucleus undergoes a dramatic transformation into a large, round pronucleus with relaxed chromatin during the brief cleavage period in metazoan embryos, enabling the activation of chromatin functions necessary for subsequent development. However, it remains unclear whether the egg cytoplasm-specific physicochemical properties play a role in pronucleus assembly. Here, we evaluated the impact of abundant RNAs in eggs on pronucleus assembly in the Xenopus laevis cell-free reconstitution system. We found that the introduction of RNAs at an appropriate concentration led to a rapid nuclear growth, more dispersed chromatin distribution, and dissociation of sperm-specific nuclear proteins from the chromatin. These chromatin remodeling properties, which were reproducible through the introduction of negatively charged compounds, facilitated the incorporation of somatic histones into chromatin in the reconstituted nuclei. Based on these findings, we propose that the remodeled chromatin by negatively charged cytoplasmic RNAs accelerates rapid decondensation of negatively charged chromatin and pronucleus assembly during the brief cleavage period following fertilization.
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