Abstract
Maternal age affects oocyte quality and early embryo development. Aberrant meiosis of oocytes and compromised early embryo development from older females could originate from defects in the nucleus, the cytoplasm, or both. Nuclear transfer has been used for decades as a tool to study nuclear-cytoplasmic interactions in early embryos, and has uncovered genomic imprinting, nuclear reprogramming, and produced animal clones. Here, we describe the technique for investigating nuclear-cyoplasmic interactions in oocytes and zygotes in female reproductive aging. Nuclear transfer can be performed efficiently and effects of the technique itself on meiosis and early embryo development are minimal as long as care is taken to minimize insult to oocytes or embryos. This protocol first focuses on use of nuclear transfer to study nucleus versus cytoplasmic origin in agingassociated meiosis defects in oocytes at the germinal vesicle (GV) stage. Then, nuclear transfer is used at the zygote stage to study nuclear and cytoplasmic abnormality and apoptosis in early development.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Keefe, D. L. (1998) Reproductive aging is an evolutionarily programmed strategy that no longer provides adaptive value. Fertil. Steril. 70, 204–206.
Navot, D., Bergh, P. A., Williams, M. A., et al. (1991) Poor oocyte quality rather than implantation failure as a cause of age-related decline in female fertility. Lancet 337, 1375–1377.
Plachot, M., Veiga, A., Montagut, J., et al. (1988) Are clinical and biological IVF parameters correlated with chromosomal disorders in early life: a multicentric study. Hum. Reprod. 3, 627–635.
Hassold, T. and Chiu, D. (1985) Maternal age-specific rates of numerical chromosome abnormalities with special reference to trisomy. Hum. Genet. 70, 11–17.
Battaglia, D. E., Goodwin, P., Klein, N. A., and Soules, M. R. (1996) Influence of maternal age on meiotic spindle assembly in oocytes from naturally cycling women. Hum. Reprod. 11, 2217–2222.
Angell, R. (1997) First-meiotic-division nondisjunction in human oocytes. Am. J. Hum. Genet. 61, 23–32.
Volarcik, K., Sheean, L., Goldfarb, J., Woods, L., Abdul-Karim, F. W., and Hunt, P. (1998) The meiotic competence of in-vitro matured human oocytes is influenced by donor age: evidence that folliculogenesis is compromised in the reproductively aged ovary. Hum. Reprod. 13, 154–160.
Hassold, T. and Hunt, P. (2001) To err (meiotically) is human: the genesis of human aneuploidy. Nat. Rev. Genet. 2, 280–291.
Wolstenholme, J. and Angell, R. R. (2000) Maternal age and trisomy-a unifying mechanism of formation. Chromosoma 109, 435–438.
McKim, K. S. and Hawley, R. S. (1995) Chromosomal control of meiotic cell division. Science 270, 1595–1601.
Janny, L. and Menezo, Y. J. (1996) Maternal age effect on early human embryonic development and blastocyst formation. Mol. Reprod. Dev. 45, 31–37.
Arbuzova, S., Hutchin, T., and Cuckle, H. (2002) Mitochondrial dysfunction and Down’s syndrome. Bioessays 24, 681–684.
Beermann, F., Hummler, E., Franke, U., and Hansmann, I. (1988) Maternal modulation of the inheritable meiosis I error Dipl I in mouse oocytes is associated with the type of mitochondrial DNA. Hum. Genet. 79, 338–340.
Nicolaidis, P. and Petersen, M. B. (1998) Origin and mechanisms of non-disjunction in human autosomal trisomies. Hum. Reprod. 13, 313–319.
Petersen, M. B. and Mikkelsen, M. (2000) Nondisjunction in trisomy 21: origin and mechanisms. Cytogenet. Cell Genet. 91, 199–203.
Liu, L. and Keefe, D. L. (2002) Ageing-associated aberration in meiosis of oocytes from senescence-accelerated mice. Hum. Reprod. 17, 2678–2685.
Liu, L. and Keefe, D. L. (2004) Nuclear origin of aging-associated meiotic defects in senescence-accelerated mice. Biol. Reprod. 71, 1724–1729.
Cui, L. B., Huang, X. Y., and Sun, E Z. (2005) Transfer of germinal vesicle to ooplasm of young mice could not rescue ageing-associated chromosome misalignmentin meiosis of oocytes from aged mice. Hum. Reprod. 20, 1624–1631.
Liu, H., Wang, C. W., Grifo, J. A., Krey, L. C., and Zhang, J. (1999) Reconstruction of mouse oocytes by germinal vesicle transfer: maturity of host oocyte cytoplasm determines meiosis. Hum. Reprod. 14, 2357–2361.
Takeuchi, T., Ergun, B., Huang, T. H., Rosenwaks, Z., and Palermo, G. D. (1999) A reliable technique of nuclear transplantation for immature mammalian oocytes. Hum. Reprod. 14, 1312–1317.
Liu, H., Zhang, J., Krey, L. C., and Grifo, J. A. (2000) In-vitro development of mouse zygotes following reconstruction by sequential transfer of germinal vesicles and haploid pronuclei. Hum. Reprod. 15, 1997–2002.
Liu, L. and Keefe, D. L. (2000) Cytoplasm mediates both development and oxidation-induced apoptotic cell death in mouse zygotes. Biol. Reprod. 62, 1828–1834.
Liu, L., Trimarchi, J. R., Smith, P. J., and Keefe, D. L. (2002) Mitochondrial dysfunction leads to telomere attrition and genomic instability. Aging Cell 1, 40–46.
Eppig, J. J. and Telfer, E. E. (1993) Isolation and culture of oocytes. Methods Enzymol. 225, 77–84.
Nagy, A., Gertsenstein, M., Vintersten, K., and Behringer, R. (2003) Manipulating the Mouse Embryo: A Laboratory Manual. Third Edition (Wassarman, P. M. and DePamphilis, M L., eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, NY: p. 192.
Lawitts, J. A. and Biggers, J. D. (1993) Culture of preimplantation embryos. Methods Enzymol. 225, 153–164.
Liu, Z. and Foote, R. H. (1995) Effects of amino acids on the development of in-vitro matured/in-vitro fertilization bovine embryos in a simple protein-free medium. Hum. Reprod. 10, 2985–2991.
Ho, Y., Wigglesworth, K., Eppig, J. J., and Schultz, R. M. (1995) Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol. Reprod. Dev. 41, 232–238.
McGrath, J. and Solter, D. (1983) Nuclear transplantation in the mouse embryo by microsurgery and cell fusion. Science 220, 1300–1302.
Liu, L., Dai, Y., and Moor, R. M. (1997) Nuclear transfer in sheep embryos: the effect of cell-cycle coordination between nucleus and cytoplasm and the use of in vitro matured oocytes. Mol. Reprod. Dev. 47, 255–264.
Barton, S. C. and Surani, M. A. (1993) Manipulations of genetic constitution by nuclear transplantation. Methods Enzymol. 225, 732–744.
Latham, K. E. and Solter, D. (1993) Transplantation of nuclei to oocytes and embryos. Methods Enzymol. 225, 719–732.
Schroeder, A. C. and Eppig, J. J. (1984) The developmental capacity of mouse oocytes that matured spontaneously in vitro is normal. Dev. Biol. 102, 493–497.
Liu, L., Ju, J. C., and Yang, X. (1998) Differential inactivation of maturation-promoting factor and mitogenactivated protein kinase following parthenogenetic activation of bovine oocytes. Biol. Reprod. 59, 537–545.
Liu, L., Oldenbourg, R., Trimarchi, J. R., and Keefe, D. L. (2000) A reliable, noninvasive technique for spindle imaging and enucleation of mammalian oocytes. Nat. Biotechnol. 18, 223–225.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Liu, L., Keefe, D.L. (2007). Nuclear Transfer Methods to Study Aging. In: Tollefsbol, T.O. (eds) Biological Aging. Methods in Molecular Biology™, vol 371. Humana Press. https://doi.org/10.1007/978-1-59745-361-5_15
Download citation
DOI: https://doi.org/10.1007/978-1-59745-361-5_15
Publisher Name: Humana Press
Print ISBN: 978-1-58829-658-0
Online ISBN: 978-1-59745-361-5
eBook Packages: Springer Protocols