Abstract
During the last decade it has been shown that most mammalian blastocysts consisted of three cell lineages. Immunofluorescence with multiple antibodies enables to identify each cell type allowing an easy detection of eventual defects. It is complementary to RT-PCR experiments as this technique allows to look at cell position and to analyze and count the proportions between the different cell types. Thus after any kind of embryo manipulation such as nuclear transfer (NT), the analysis of the three cell lineages by immunofluorescence will provide criteria for good or poor development.
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References
Stephenson RO, Rossant J, Tam PP (2012) Intercellular interactions, position, and polarity in establishing blastocyst cell lineages and embryonic axes. Cold Spring Harb Perspect Biol 4. doi:10.1101/cshperspect.a008235
Chazaud C, Yamanaka Y, Pawson T et al (2006) Early lineage segregation between epiblast and primitive endoderm in mouse blastocysts through the Grb2-MAPK pathway. Dev Cell 10:615–624
Plusa B, Piliszek A, Frankenberg S et al (2008) Distinct sequential cell behaviours direct primitive endoderm formation in the mouse blastocyst. Development 135:3081–3091
Guo G, Huss M, Tong GQ et al (2010) Resolution of cell fate decisions revealed by single-cell gene expression analysis from zygote to blastocyst. Dev Cell 18:675–685
Frankenberg S, Gerbe F, Bessonnard S et al (2011) Primitive endoderm differentiates via a three-step mechanism involving Nanog and RTK signaling. Dev Cell 21:1005–1013
Kang M, Piliszek A, Artus J, Hadjantonakis AK (2013) FGF4 is required for lineage restriction and salt-and-pepper distribution of primitive endoderm factors but not their initial expression in the mouse. Development 140:267–279
Sebastiano V, Gentile L, Garagna S et al (2005) Cloned pre-implantation mouse embryos show correct timing but altered levels of gene expression. Mol Reprod Dev 70:146–154
Jouneau A, Zhou Q, Camus A et al (2006) Developmental abnormalities of NT mouse embryos appear early after implantation. Development 133:1597–1607
Nagy A, Gertsenstein M, Vintersten K et al (2003) Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, NY
Acknowledgement
This work was supported by the ANR “EpiNodal.”
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Chauveau, S., Chazaud, C. (2015). Assessment of Cell Lineages and Cell Death in Blastocysts by Immunostaining. In: Beaujean, N., Jammes, H., Jouneau, A. (eds) Nuclear Reprogramming. Methods in Molecular Biology, vol 1222. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1594-1_13
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DOI: https://doi.org/10.1007/978-1-4939-1594-1_13
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Publisher Name: Humana Press, New York, NY
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