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DNA synthesis, microtubule and nuclear dynamics in porcine parthenotes

Published online by Cambridge University Press:  26 September 2008

Lin Liu ,
Caroline Lee  and
Robert M. Moor
Lin Liu*
Affiliation:
Department of Development and Genetics, The Babraham Institute, Cambridge CB2 4AT, UK
Caroline Lee
Affiliation:
Department of Development and Genetics, The Babraham Institute, Cambridge CB2 4AT, UK
Robert M. Moor
Affiliation:
Department of Development and Genetics, The Babraham Institute, Cambridge CB2 4AT, UK
*
Lin Liu Animal Science Department, University of Connecticut, 3636 Horsebarn Road, Ext U-40, Storrs, CT 06269, USA. Fax: +1(860) 486-4375. e-mail: LLIU@anscil.cag.uconn.edu.
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Summary

Parthenogenetically activated mammalian oocytes have been used in the past decade as cytoplasts, in an attempt to support the development of nuclear transplant embryos. The present experiments were undertaken to study the DNA synthesis and the organisation of microtubules, nuclear envelope and chromatin during the first cell cycle of electrically activated porcine oocytes (parthenotes) matured in vitro by using immunocytochemistry and laser scanning confocal microscopy. The results showed that pronuclear-like (PN) formation began 4–5 h post-activation (hpa), whilst DNA synthesis as revealed by bromodeoxyuridine incorporation was initiated 5–6 hpa, with a maximum number of labelled oocytes (73%) around 11 hpa, and persisted in some parthenotes until 15–16 hpa. In the metaphase II (MII) oocytes, microtubules were detected only in the metaphase II spindle; no lamin A/C antigen was observed. Electrical DC pulses resulted in 91% of MII oocytes being activated and confocal microscopy Indicated that microtubules were assembled in the spindle first for the extrusion of a second polar body, and for the second time for division from one to two cells. Nuclear envelope, indicated by anti-lamin A/C stain, was formed around the time of PN formation and surrounded the nuclear chromatin of 1- and 2-cell parthenogenotea. These results demonstrate that the apparent normality in both DNA synthesis and dynamics of microtubules and nuclear envelope is involved with chromosomal organisation in the parthenotes. In addition, the use of electrically activated IVM oocytes for both nuclear transfer and parthenogenetic studies in pigs is discussed.

Keywords

ActivationCytoskeletonDNA synthesisIVM oocytePorcine
Type
Article
Information
Zygote , Volume 4 , Issue 2 , May 1996 , pp. 139 - 144
Copyright
Copyright © Cambridge University Press 1996

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