Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development

  • Jinling Liu
  • Aimin LiuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2047)


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.


Immunohistochemistry RNA in situ hybridization Cryosection Antibody Digoxigenin Fluorescent 



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 Department of Biological Sciences, The University of Iowa, Iowa City, IA 52242, USA. Research in the Liu lab has been supported by US NSF (IOS-0949877 and IOS-1257540), US NIH (HD083625) and a Penn State University new faculty start-up fund.


  1. 1.
    Herculano-Houzel S, Mota B, Lent R (2006) Cellular scaling rules for rodent brains. Proc Natl Acad Sci U S A 103(32):12138–12143. Scholar
  2. 2.
    Gall JG, Pardue ML (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A 63(2):378–383CrossRefGoogle Scholar
  3. 3.
    Komminoth P, Merk FB, Leav I, Wolfe HJ, Roth J (1992) Comparison of 35S- and digoxigenin-labeled RNA and oligonucleotide probes for in situ hybridization. Expression of mRNA of the seminal vesicle secretion protein II and androgen receptor genes in the rat prostate. Histochemistry 98(4):217–228CrossRefGoogle Scholar
  4. 4.
    Wilcox JN (1993) Fundamental principles of in situ hybridization. J Histochem Cytochem 41(12):1725–1733CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Biology, Center for Cellular Dynamics, Eberly College of Science, Huck Institute of Life SciencesThe Penn State UniversityPennsylvaniaUSA

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