Skip to main content

Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2047))

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.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   299.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  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. https://doi.org/10.1073/pnas.0604911103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  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):378–383

    Article  CAS  Google Scholar 

  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):217–228

    Article  CAS  Google Scholar 

  4. Wilcox JN (1993) Fundamental principles of in situ hybridization. J Histochem Cytochem 41(12):1725–1733

    Article  CAS  Google Scholar 

Download references

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 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.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Aimin Liu .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Liu, J., Liu, A. (2020). Immunohistochemistry and RNA In Situ Hybridization in Mouse Brain Development. In: Sprecher, S. (eds) Brain Development. Methods in Molecular Biology, vol 2047. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9732-9_27

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9732-9_27

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9731-2

  • Online ISBN: 978-1-4939-9732-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics