Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-25T13:11:13.891Z Has data issue: false hasContentIssue false
  • English
  • Français

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: [email protected].
Get access Rights & Permissions [Opens in a new window]

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Almeida, P.A. & Bolton, V.N. (1995). The effect of temperature fluctuations on the cytoskeletal organisation and chromosomal constitution of the human oocyte. Zygote 3, 357–65.CrossRefGoogle ScholarPubMed
Barnes, F.L., Collas, P., Powell, R., King, W.A., Westhusin, M., Shepherd, D. 1993. Influence of recipient oocyte cell cycle state on DNA synthesis, nuclear envelope breakdown, Chromosome constitution, and development in nuclear transplant bovine embryos. Mol. Reprod. Dev. 36, 33–41.CrossRefGoogle Scholar
Campbell, K.H.S., Loi, P., Cappai, P. & Wilmut, I. (1994). Improved development to blastocyst of ovine nuclear transfer embryos reconstructed during the presumptive Sphase of enucleated activated oocytes. Biol. Reprod. 50, 1385–93.CrossRefGoogle ScholarPubMed
Collas, P. & Robl, J.M. (1990). Factors affecting the efficiency of nuclear transplantation in the rabbit embryos. Biol. Reprod. 43, 877–84.CrossRefGoogle Scholar
Kim, N.-H., Funahashi, H., Stumpf, T.T. & Day, B.N. (1994). Microtubule organization of porcine oocytes during in vitro fertilization. Blot. Reprod. 50 (Suppl), 146.Google Scholar
Kim, N.-H., Funahashi, H., Prather, R.S., Schatten, G. & Day, B.N. (1996 a). Microtubule and microfilament dynamics in porcine oocytes during meiotic maturation. Mol. Reprod. Dev. 43, 248–55.3.0.CO;2-#>CrossRefGoogle ScholarPubMed
Kim, N.-H., Moon, S.J., Prather, R.S. & Day, B.N. (1996 b). Cytoskeletal alteration in aged porcine oocytes and parthenogenesis. Mol. Reprod. Dev. 43, 513–18.3.0.CO;2-#>CrossRefGoogle ScholarPubMed
Landa, V. & Kopecny, V. (1995). DNA synthesis and distribution in parthenogenetic bovine embryos. Theriogenology 43, 871–81.CrossRefGoogle ScholarPubMed
Laurincik, J., Hyttel, P. & Kopecny, V. (1995). DNA synthesis and pronucleus development in pig zygotes obtained in vivo: an autoradiographic and ultrastructural study. Mol. Reprod. Dev. 40, 325–32.Google ScholarPubMed
Liu, L. & Moor, R.M. (1995). Optimisation of electrical activation of porcine oocytes matured in vitro. J. Reprod. Fertil., Abstract Series. 15, 70.Google Scholar
Liu, L., Moor, R.M., Laurie, S. & Notarianni, E. (1995). Nuclear remodelling and early development in cryopreserved, porcine primordial germ cells following nuclear transfer into in vitro-matured oocyted. Int. J. Dev. Biol. 39, 639–44.Google Scholar
Long, C.R., Pinto-Correia, C., Duby, R.T., Ponce, de Leon F.A., Boland, M.P., Roche, J.F. & Robl, J.M. (1993). Chromatin and microtubule morphology during the first cell cycle in bovine zygotes. Mol. Reprod. Dev. 36, 23–32.CrossRefGoogle ScholarPubMed
Machaty, Z., Mayers, M.A. & Prather, R.S. (1995). Parthenogenetic activation of porcine oocytes with guanosine-5'-O- (3'-thiotriphosphate). Biol. Reprod. 52, 753–8.CrossRefGoogle ScholarPubMed
Mayes, M.A., Stogsdill, P.L. & Prather, R.S. (1995). Parthenogenetic activation of pig oocytes by protein kinase inhibition. Biol. Reprod. 53, 270–5.CrossRefGoogle Scholar
Moor, R.M. & Trounson, A.O. (1977). Hormonal and follicular factors affecting maturation of sheep oocytes in vitro and their subsequent developmental capacity. J. Reprod. Fertil. 49, 101–9.CrossRefGoogle ScholarPubMed
Navara, C.S., First, N.L. & Schatten, G. (1994). Microtubule organization in the cow during fertilization, polyspermy, parthenogenesis, and nuclear transfer the role of the sperm aster. Dev. Biol. 162, 29–40.CrossRefGoogle ScholarPubMed
Nussbaum, A.J. & Prather, R.S. (1995). Differential effects of protein synthesis inhibitors on porcine oocyte activation. Mol. Reprod. Dev. 41, 70–5.CrossRefGoogle ScholarPubMed
Ozil, J.P. (1990). The parthenogenetic development of rabbit oocytes after repetitive pulsatile stimulation. Development 109, 117–27.CrossRefGoogle Scholar
Pinto-Correia, C., Collas, P., Ponce, de Leon F.A., & Robl, J.M. (1993). Chromatin and microtubule organization in the first cell cycle in rabbit parthenotes and nuclear transplant embryos. Mol. Reprod. Dev. 34,33–42.CrossRefGoogle ScholarPubMed
Prather, R.S., Sims, M.M., Maul, G.G., First, N.L. & Schatten, G. (1989). Nuclear lamin antigens are developmentally regulated during porcine and bovine embryogenesis. Blol. Reprod. 40, 123–32.CrossRefGoogle Scholar
Smith, L.C. & Wilmut, I. (1989). Influence of nuclear and cytoplasmic activity on development in vivo of sheep embryos after nuclear transplantation. Biol. Reprod. 40, 1027–35.CrossRefGoogle ScholarPubMed
Sun, F.Z., Hoyland, J., Huang, X., Mason, W. & Moor, R.M. (1992). A comparison of intracellular changes in porcine eggs after fertilization and electroactivation. Development 115, 947–56.CrossRefGoogle ScholarPubMed