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Nucleolar-based Dux repression is essential for embryonic two-cell stage exit.

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Robust ribosomal RNA synthesis and nucleolar maturation are essential for mammalian embryo development. Disrupting nucleoli in embryonic stem cells and embryos hinders the silencing of MERVL-driven genes, impacting early development.

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Area of Science:

  • Developmental Biology
  • Genetics
  • Cell Biology

Background:

  • Mammalian embryonic development requires a transition from maternal to embryonic gene expression.
  • The two-cell (2C) stage in mice involves transient upregulation of MERVL transposons and 2C genes.
  • The mechanisms regulating 2C gene expression and subsequent silencing are not fully understood.

Purpose of the Study:

  • To investigate the role of nucleolar maturation and ribosomal RNA (rRNA) synthesis in exiting the 2C state.
  • To understand how nucleolar function impacts the regulation of MERVL-driven genes and the activator Dux.
  • To determine the necessity of nucleolar maturation for embryonic developmental progression.

Main Methods:

  • Utilized mouse embryonic stem cells (ESCs) and early mouse embryos.
  • Manipulated RNA polymerase I activity to disrupt rRNA synthesis and nucleolar maturation.
  • Assessed nucleolar structure, rRNA output, and Dux protein localization.
  • Analyzed gene expression patterns and developmental progression.

Main Results:

  • Immature nucleoli with reduced rRNA output were observed in 2C-like cells and 2-cell embryos.
  • Disruption of nucleolar maturation in ESCs enhanced conversion to a 2C-like state by detaching Dux.
  • In embryos, nucleolar disruption led to impaired Dux silencing, failed nucleolar maturation, and arrest at the 2- to 4-cell stage.

Conclusions:

  • Robust rRNA synthesis and nucleolar maturation are critical for exiting the 2C developmental stage.
  • The nucleolus plays a key role in repressing MERVL-driven transcription, including the Dux activator.
  • Proper nucleolar function is essential for successful early mammalian embryonic development.