In vitro fertilisation of mouse oocytes reconstructed by transfer of metaphase II chromosomes results in live births

Zygote. 2001 Feb;9(1):9-14. doi: 10.1017/s0967199401001022.

Abstract

The interaction between nucleus and cytoplasm can be explored through nuclear transfer. We describe here another tool to investigate this interaction: MII meiotic apparatus transfer (MAT) between mouse oocytes. In this study, the MII oocyte meiotic apparatus or spindle from C57BL/6 mice, a black strain, was transferred into an enucleated metaphase oocyte from Kunming mouse, a white strain. The results showed that the enucleation rate by treating oocytes with 3% sucrose was 100%, but the electrofusion efficiency was very low, with only 17.6% of reconstructed karyoplast-recipient cytoplasm pairs fused. When the fused oocytes were exposed to spermatozoa from C57BL/6 mice, 9 of 11 (82%) were fertilised. Eight reconstructed embryos at 1- to 4-cell stages were transferred into the oviducts of two synchronously pregnant Kunming strain fosters and one delivered two normal C57BL/6 offspring. This study indicates that MII meiotic apparatus or spindle sustains normal structure and function after micromanipulation and electrofusion. MAT provides a model for further research on the application of this technique to assisted human reproduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromosomes*
  • Female
  • Fertilization in Vitro / methods*
  • Gene Transfer Techniques*
  • Male
  • Meiosis
  • Metaphase / genetics
  • Mice
  • Mice, Inbred C57BL
  • Micromanipulation
  • Oocytes / drug effects
  • Oocytes / physiology*
  • Pregnancy
  • Pregnancy Outcome
  • Spindle Apparatus / genetics
  • Sucrose / pharmacology

Substances

  • Sucrose