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Nucleolar changes in bovine nucleotransferred embryos.

V Baran¹, X Vignon, D LeBourhis

¹Institut National de la Recherche Agronomique, Biologie du Développement et Biotechnologies, 78352 Jouy-en-Josas, France.

Biology of Reproduction

|January 24, 2002

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Nuclear reprogramming in cloned bovine embryos is essential for development. Nucleolar changes, including rRNA gene expression, indicate successful reprogramming in embryonic cloning versus somatic cloning.

Area of Science:

Reproductive biology
Cell biology
Developmental biology

Background:

Nucleolar structure and function are critical for cell viability and proliferation.
Nuclear reprogramming is a key challenge in somatic cell nuclear transfer (SCNT) for successful embryonic development.

Purpose of the Study:

To investigate nucleolar changes during early development of bovine embryos reconstructed via SCNT.
To compare nucleolar dynamics between embryonic and somatic cell cloning in cattle.
To assess the correlation between nucleolar morphology and developmental potential.

Main Methods:

Embryos were reconstructed using enucleated oocytes and blastomeres or fetal fibroblasts.
Nucleotransferred (NT) embryos were collected at various developmental stages (1-cell to 16-cell).

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Immunocytochemistry, light and electron microscopy, and DNA localization techniques (TdT, nick-translation) were employed.

Main Results:

Nucleoli transformed from active reticulated structures to inactive bodies during early cleavage stages in both cloning types.
Reactivation of nucleoli, marked by vacuole reappearance and chromatin association, occurred by the late 8-cell stage.
Embryonic cloning showed more robust nucleolar rebuilding and earlier reactivation compared to somatic cloning.
Somatic cloned embryos exhibited delayed nucleolar morphological changes and reduced developmental tendency.

Conclusions:

Complete reprogramming of rRNA gene expression is integral to nuclear reprogramming for bovine SCNT development.
Nucleolar morphology serves as a sensitive indicator of nuclear reprogramming efficiency.
Embryonic cloning demonstrates superior developmental potential, likely due to more effective nuclear reprogramming.