Detection of DNA Double-Stranded Breaks in Mouse Oocytes

Chrysoula Zorzompokou; Marios Ipeirotis; Marios Kaisar Martzoukos; Petros Marangos

doi:10.3791/65494

Journal

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Developmental Biology

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Detection of DNA Double-Stranded Breaks in Mouse Oocytes

JoVE Journal
Developmental Biology
Author Produced

This content is Free Access.

JoVE Journal Developmental Biology

Detection of DNA Double-Stranded Breaks in Mouse Oocytes

2,136 Views

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07:46 min

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June 23, 2023

DOI:

10.3791/65494-v

DOI:

10.3791/65494-v

•

07:46 min

June 23, 2023

•

18 Views

Chrysoula Zorzompokou^1,2, Marios Ipeirotis^1,2, Marios Kaisar Martzoukos¹, Petros Marangos^1,2

¹Department of Biological Applications and Technology,University of Ioannina, ²Biomedical Research Institute,Foundation for Research and Technology

Transcript

Our research aims to understand how the cell cycle is regulated in mammalian oocytes and in preimplantation embryos. A key aspect of our research involves examination of repair capacity and DNA damage response in different maturation states is in oocytes and also in preimplantation embryos. We are particularly interested in understanding how the checkpoints respond and which repair mechanisms establish after potential DNA damage.

Our lab was the first to mechanistically examine the DNA damage response in mammalian oocytes. We have shown the absence of the G2 prophase DNA damage checkpoint in mammalian oocytes. And we observed the importance of the spindle assembly checkpoint in establishing metaphase arrest after damage.

By using this protocol, it is possible to detect DNA double strand breaks with accuracy and to determine whether oocytes have DNA damage and whether DNA damage is being repaired. This protocol involves the use of immunofluorescence which allows us to identify the exact sites of damage to DNA.

Summary

Maintaining oocyte genome integrity is necessary to ensure the genetic fidelity in the resulting embryo. Here, we present an accurate protocol for detecting DNA double-strand breaks in mammalian female germ cells.