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Methods for Precisely Localized Transfer of Cells or DNA into Early Postimplantation Mouse Embryos

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Published on: December 25, 2015

Asynchronous DNA replication and origin licensing in the mouse one-cell embryo.

Yasuhiro Yamauchi¹, Monika A Ward, W Steven Ward

¹Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96822, USA.

Journal of Cellular Biochemistry

|March 21, 2009

Summary

This summary is machine-generated.

Mammalian one-cell embryos regulate DNA replication licensing. Oocyte cytoplasm permits de novo licensing for 3 hours post-activation and controls asynchronous DNA synthesis between pronuclei.

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Area of Science:

Cell Biology
Developmental Biology
Genetics

Background:

Metazoan DNA replication requires origin licensing during G1 to prevent duplication.
Oocyte cytoplasm inhibits new licensing during S phase, ensuring replication fidelity.
Mammalian one-cell embryos present a unique system for studying DNA replication due to separate pronuclei within a single cytoplasm.

Purpose of the Study:

To determine the duration of oocyte cytoplasm's ability to support licensing after activation.
To investigate the regulation of DNA synthesis initiation and progression in the mammalian one-cell embryo.
To explore the cytoplasmic control over asynchronous DNA replication in pronuclei.

Main Methods:

Injected mouse sperm halos into parthenogenetically activated oocytes to assess de novo licensing timing.
Transferred pronuclei between zygotes at different cell cycle phases (G1 and S) using intracytoplasmic sperm injection (ICSI).
Monitored DNA replication initiation and progression in pronuclei within manipulated oocytes.

Main Results:

De novo licensing occurred up to 3 hours post-oocyte activation, preceding DNA replication by 4 hours.
Oocyte cytoplasm supported asynchronous DNA synthesis initiation in the two pronuclei, with a minimum 2-hour difference.
Paternal pronuclei transferred into S phase ooplasm did not prematurely replicate, and those transferred into G1 ooplasm continued replication, indicating cytoplasmic regulation.

Conclusions:

The mammalian one-cell embryo serves as a valuable model for understanding DNA synthesis regulation.
Oocyte cytoplasm plays a critical role in controlling the timing and progression of DNA replication.
Cell cycle progression is tightly regulated by cytoplasmic factors in early mammalian development.