Abstract
A common problem in research laboratories that study the mammalian embryo is the limited supply of live material. For this reason, new methods are constantly being developed and existing methods for in vitro models using cells in culture are being adapted to represent embryogenesis. Three-dimensional fluorescence in situ hybridization (3D-FISH) is an important tool to study where genomic sequences are positioned within nuclei without interfering with this 3D organization. When used in the embryo, this technique provides vital information about the distribution of specific sequences in relation to embryonic nuclear substructures such as nucleolar precursor bodies and chromocenters. In this chapter, we will present a detailed description of FISH in order to perform 3D-FISH in the early preimplantation murine embryos.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Halling, K. C., Kipp, B. R. (2007) Fluorescence in situ hybridization in diagnostic cytology. Hum Pathol 38, 1137–1144.
Sreekantaiah, C. (2007) FISH panels for hematologic malignancies. Cytogenet Genome Res 118, 284–296.
Foster, H. A., Bridger, J. M. (2005) The genome and the nucleus: a marriage made by evolution. Genome organisation and nuclear architecture. Chromosoma 114, 212–229.
Walter, J., Joffe, B., Bolzer, A., Albiez, H., Benedetti, P. A., Muller, S., Speicher, M. R., Cremer, T., Cremer, M., Solovei, I. 2006 Towards many colors in FISH on 3D-preserved interphase nuclei. Cytogenet Genome Res 114, 367–378.
Kupper, K., Kolbl, A., Biener, D., Dittrich, S., von Hase, J., Thormeyer, T., Fiegler, H., Carter, N. P., Speicher, M. R., Cremer, T., Cremer, M. (2007) Radial chromatin positioning is shaped by local gene density, not by gene expression. Chromosoma 116, 285–306.
Maalouf, W. E., Liu, Z., Brochard, V., Renard, J. P., Debey, P., Beaujean, N., Zink, D. (2009) Trichostatin A treatment of cloned mouse embryos improves constitutive heterochromatin remodeling as well as developmental potential to term. BMC Dev Biol 9, 11.
Werner, M., Wilkens, L., Aubele, M., Nolte, M., Zitzelsberger, H., Komminoth, P. (1997) Interphase cytogenetics in pathology: principles, methods, and applications of fluorescence in situ hybridization (FISH). Histochem Cell Biol 108, 381–390.
Wassarman, P. M. (2005) Contribution of mouse egg zona pellucida glycoproteins to gamete recognition during fertilization. J Cell Physiol 204, 388–391.
Bouzinba-Segard, H., Guais, A., Francastel, C. (2006) Accumulation of small murine minor satellite transcripts leads to impaired centromeric architecture and function. Proc Natl Acad Sci U S A 103, 8709–8714.
Acknowledgments
We thank Pierre Adenot for technical assistance on the confocal microscope platform MIMA2 (Microscopie et Imagerie des Microorganismes, Animaux et Elements) and UEAR for animal care. We are also grateful to Eve Devinoy (INRA) and Claire Francastel (Institut Cochin) for their help. This work was supported by INRA grant “Crédits incitatifs PHASE”.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Maalouf, W.E., Aguirre-Lavin, T., Herzog, L., Bataillon, I., Debey, P., Beaujean, N. (2010). Three-Dimensional Fluorescence In Situ Hybridization in Mouse Embryos Using Repetitive Probe Sequences. In: Bridger, J., Volpi, E. (eds) Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology, vol 659. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-789-1_31
Download citation
DOI: https://doi.org/10.1007/978-1-60761-789-1_31
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-788-4
Online ISBN: 978-1-60761-789-1
eBook Packages: Springer Protocols