Abstract
The use of flow cytometry in vertebrate nervous tissues is hampered by the morphological complexity and high level of interconnectivity intrinsic to their cellular constituents. Here we describe a simplified procedure for the identification and quantitative analysis of neural cells by flow cytometry based on the isolation and immunolabeling of fresh cell nuclei . We have applied this procedure for the quantitative analysis of apoptosis and DNA synthesis in the embryonic brain .
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References
Westra JW, Rivera RR, Bushman DM et al (2010) Neuronal DNA content variation (DCV) with regional and individual differences in the human brain. J Comp Neurol 518:3981–4000
López-Sánchez N, Frade JM (2013) Cell cycle analysis in the vertebrate brain using immunolabeled fresh cell nuclei. Bio-Protocol 3:e973. http://www.bio-protocol.org/e973
De la Rosa EJ, de Pablo F (2000) Cell death in early neural development: beyond the neurotrophic theory. Trends Neurosci 23:454–458
Wang W, Bu B, Xie M et al (2009) Neural cell cycle dysregulation and central nervous system diseases. Prog Neurobiol 89:1–17
López-Sánchez N, Frade JM (2013) Genetic evidence for p75NTR-dependent tetraploidy in cortical projection neurons from adult mice. J Neurosci 33:7488–7500
Nunez R (2001) DNA measurement and cell cycle analysis by flow cytometry. Curr Issues Mol Biol 3:67–70
Acknowledgements
This study was supported by grants from the “Ministerio de Ciencia e Innovación” (SAF2012-38316) and “Fundación Areces” (CIVP16A1815).
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López-Sánchez, N., Frade, J.M. (2015). Flow Cytometric Analysis of DNA Synthesis and Apoptosis in Central Nervous System Using Fresh Cell Nuclei . In: Lossi, L., Merighi, A. (eds) Neuronal Cell Death. Methods in Molecular Biology, vol 1254. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2152-2_3
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DOI: https://doi.org/10.1007/978-1-4939-2152-2_3
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-2152-2
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