Abstract
During development, the mouse brain is progressively divided into functionally distinct compartments. Numerous neuronal and glial cell types are subsequently generated in response to various inductive signals. Each cell expresses a unique combination of genes encoding proteins from transcription factors to neurotransmitters that define its role in brain function. To understand these important and highly sophisticated processes, it is critical to accurately locate the various proteins and cells that produce them. In this chapter, we introduce the techniques of immunohistochemistry, which detects the localization of specific proteins, and RNA in situ hybridization, which enables the visualization of specific mRNAs.
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References
Kandel E, Schwartz J, Jessell T (2000) Principles of neural science, 4th edn. McGraw-Hill, Health Professions Division, New York, NY. ISBN 0-8385-7701-6
Joseph G. Gall, Mary Lou Pardue (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations*. Proc Natl Acad Sci USA 63:378–383
Komminoth P, Merk FB, Wolfe HJ, Roth J (1992) Comparison of 35S-and digoxigenin-labeled RNA and oligonucleotide probes for in situ hybridization. Histochemistry 98: 217–228
Wilcox JN (1993) Fundamental principles of in situ hybridization. J Histochem Cytochem 41:1725–1733. doi:10.1177/41.12.8245419
Acknowledgments
We would like to thank Dr. Simeone for providing the RNA in situ probe for Otx2. The monoclonal antibody against Pax6 developed by Dr. Jessell was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of Biological Sciences, Iowa City, IA 52242. A. L. is supported by an NSF grant (IOS-0949877) and a Penn State new lab start-up fund.
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Liu, J., Liu, A. (2014). Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development. In: Sprecher, S. (eds) Brain Development. Methods in Molecular Biology, vol 1082. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-655-9_18
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DOI: https://doi.org/10.1007/978-1-62703-655-9_18
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-654-2
Online ISBN: 978-1-62703-655-9
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