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Assessment of the cellular DNA content of whole mounted mouse and human oocytes and of blastomeres containing single or multiple nuclei

Published online by Cambridge University Press:  26 September 2008

Nicola J. Winston ,
Martin H. Johnson  and
Peter R. Braude
Nicola J. Winston
Affiliation:
Department of Anatomy, University of Cambridge, University of Cambridge, and Department of Obstetrics and Gynaecology, St Thomas' Hospital, London, UK
Martin H. Johnson*
Affiliation:
Department of Anatomy, University of Cambridge, University of Cambridge, and Department of Obstetrics and Gynaecology, St Thomas' Hospital, London, UK
Peter R. Braude
Affiliation:
Department of Anatomy, University of Cambridge, University of Cambridge, and Department of Obstetrics and Gynaecology, St Thomas' Hospital, London, UK
*
Dr M.H. Johnson, Embryo and Gamete Research Group, Department of Anatomy, University of Cambridge, Downing Street, Cambridge CB2 3DY, UK. Tel: (0223) 333789. Fax: (0223) 333786.
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Summary

The nuclear DNA content of intact, live or fixed, human and mouse oocytes and blastomeres has been measured rapidly and reliably. Chromosomal DNA has been stained with DAPI, the fluorescent emission from which has been measured photocytometrically. In vitro fertilised mouse oocytes and embryos at various stages of development were assessed for their DNA content. The mean values of 1C, 2C and 4C DNA content were clearly different, and it was possible to assign correctly individual values for DNA content to each class with 92%, 61% and 81% confidence respectively. Maintaining the cells as whole mounts allowed other morphological and structural features to be examined. When formation of multiple micronuclei was induced in mouse oocytes by their insemination in the presence of nocodazole, the additive signal from all the micronuclei in one zygote was equivalent to the expected DNA content. Application to early human blastomeres of this photocytometric technique for measurement of the total cellular DNA content revealed that multinucleated blastomeres contained 2C to 4C DNA levels, consistent with a diploid DNA content.

Keywords

Cellular DNA contentEmbryoHumanMouseMultinucleationOocyte
Type
Article
Information
Zygote , Volume 1 , Issue 1 , February 1993 , pp. 17 - 25
Copyright
Copyright © Cambridge University Press 1993

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