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Foster, Helen A.; Sturmey, Roger G.; Stokes, Paula J.; Leese, Henry J.; Bridger, Joanna M.; Griffin, Darren K. (2010)
Publisher: Springer
Languages: English
Types: Part of book or chapter of book
Subjects: Q
Insight into the normal and abnormal function of an interphase nucleus can be revealed by using fluorescence in situ hybridization (FISH) to determine chromosome copy number and/or the nuclear position of loci or chromosome territories. FISH has been used extensively in studies of mouse and human early embryos, however, translation of such methods to domestic species have been hindered by the presence of high levels of intracytoplasmic lipid in these embryos which can impede the efficiency of FISH. This chapter describes in detail a FISH protocol for overcoming this problem. Following extensive technical development, the protocol was derived and optimized for IVF porcine embryos to enable investigation of whole chromosome and subchromosomal regions by FISH during these early stages of development. Porcine embryos can be generated in-vitro using semen samples from commercial companies and oocytes retrieved from discarded abattoir material. According to our method, porcine embryos are lyzed and immobilized on slides using Hydrochloric acid and "Tween 20" detergent, prior to pretreatment with RNase A and pepsin before FISH. The method described has been optimized for subsequent analysis of FISH in two dimensions since organic solvents, which are necessary to remove the lipid, have the effect of flattening the nuclear structure. The work in this chapter has focussed on the pig; however, such methods could be applied to bovine, ovine, and canine embryos, all of which are rich in lipid.
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    • 1. Hanel ML, Wevrick R (2001). The role of genomic imprinting in human developmental disorders: lessons from Prader-Willi syndrome. Clin Genet, 59, 156-164
    • 2. McKenzie LJ, Carson SA, Marcelli S, Rooney E, Cisneros P, Torskey S, Buster J, Simpson JL, Bischoff FZ. (2004). Nuclear chromosomal localization in human preimplantation embryos: correlation with aneuploidy and embryo morphology. Hum Reprod, 19, 2231-2237.
    • 3. Diblík J, Macek M Sr, Magli MC, Krejcí R, Gianaroli L. (2007) Chromosome topology in normal and aneuploid blastomeres from human embryos. Prenat Diagn, 27, 1091-1099.
    • 4. Finch KA, Fonseka G, Ioannou D, Hickson N, Barclay Z, Chatzimeletiou K, Mantzouratou A, Handyside A, Delhanty J, Griffin DK. (2008). Nuclear organisation in totipotent human nuclei and its relationship to chromosomal abnormality. J Cell Sci, 121, 655-663.
    • 5. Lanctôt C, Kaspar C, Cremer T. (2007). Positioning of the mouse Hox gene clusters in the nuclei of developing embryos and differentiating embryoid bodies. Exp Cell Res, 313, 1449-1459.
    • 6. Koehler D, Zakhartchenko V, Froenicke L, Stone G, Stanyon R, Wolf E, Cremer T, Brero A. (2009). Changes of higher order chromatin arrangements during major genome activation in bovine preimplantation embryos. Exp Cell Res, 315, 2053-2063.
    • 7. Lunney JK. (2007) Advances in swine biomedical model genomics. Int J Biolol Sci, 3, 179.
    • 8. Weiss RA. (1998) Transgenic pigs and virus adaptation. Nature, 391; 327-327.
    • 9. Guan TY, Holley RA (2003) Pathogen Survival in Swine Manure Environments and Transmission of Human Enteric Illness-A Review Sponsoring organizations: Manitoba Livestock Manure Management Initiative and Manitoba Rural Adaptation Council. In.: Am Soc Agronom, 383-392.
    • 10. Rettenberger G, Klett C, Zechner U, Kunz J, Vogel W, Hameister H. (1995) Visualization of the conservation of synteny between humans and pigs by heterologous chromosomal painting. Genomics, 26, 372-378.
    • 11. Lim D, Cho YM, Lee KT, Kang Y, Sung S, Nam J, Park EW, Oh SJ, Im SK, Kim H. (2009). The Pig Genome Database (PiGenome): an integrated database for pig genome research. Mamm Genome, 20, 60-66.
    • 12. Abeydeera LR, Day BN. (1997) Fertilization and subsequent development in vitro of pig oocytes inseminated in a modified tris-buffered medium with frozen-thawed ejaculated spermatozoa. Biol Reprod, 57, 729-734.
    • 13. Petters RM, Reed ML. (1991) Addition of taurine or hypotaurine to culture medium improves development of one and two cell pig embryos in vitro. Theriogenology, 35, 253.
    • 14. Sturmey RG, Leese HJ. (2003) Energy metabolism in pig oocytes and early embryos. Reproduction, 126, 197-204.
    • 15. Rooney DE, Czepulkowski BH. (1986) Human Cytogenetics: A Practical Approach. Oxford, IRL Press.
    • 16. Abeydeera LR, Wang WH, Cantley TC, Rieke A, Murphy CN, Prather RS, Day BN. (2000) Development and viability of pig oocytes matured in a protein-free medium containing epidermal growth factor. Theriogenology, 54, 787-797.
    • 17. Jeong BS, Yang X. (2001) Cysteine, glutathione, and Percoll treatments improve porcine oocyte maturation and fertilization in vitro. Mol Reprod Dev, 59, 330-335.
    • 18. Petters RM, Johnson BH, Reed ML, Archibong AE. (1990) Glucose, glutamine and inorganic phosphate in early development of the pig embryo in vitro. J Reprod Fertil, 89, 269-275.
    • 19. Coonen E, Dumoulin JC, Ramaekers FC, Hopman AH. (1994) Optimal preparation of preimplantation embryo interphase nuclei for analysis by fluorescence in-situ hybridisation. Hum Reprod, 9, 533-537.
    • 20. Harper JC, Dawson K, Delhanty JD, Winston RM. (1995) The use of fluorescent in-situ hybridisation (FISH) for the analysis of invitro fertilization embryos: a diagnostic tool for the infertile couple. Hum Reprod, 10, 3255-3258.
    • 21. Xu K, Huang T, Liu T, Shi Z, Rosenwaks Z. (1998) Improving the fixation method for preimplantation genetic diagnosis by fluorescent in situ hybridisation. J Assist Reprod Genet, 15, 570-574.
    • 22. Daphnis DD, Delhanty JD, Jerkovic S, Geyer J, Craft I, Harper JC. (2005) Detailed FISH analysis of day 5 human embryos reveals the mechanisms leading to mosaic aneuploidy. Hum Reprod, 20, 129-137.
    • 23. van Minnen J, van Kesteren RE. (1999) Methods towards detection of protein synthesis in dendrites and axons. In: Modern Techniques in Neuroscience Research (Windhorst U, Johansson H, eds.), Chapter 3, pp. 57-88. Springer-Verlag, Heidelberg, Germany.
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