Skip to main content

Neuropeptide RNA Localization in Tissue Sections

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 789))

Abstract

In situ hybridization has become a routine technique to provide insights into RNA localization. However, different protocols exist for multiple purposes, and it is, therefore, important to clearly define specific needs to choose the most suitable one(s). For instance, in situ hybridization can target different types of RNA, including mRNA or small noncoding RNA such as micro RNA (miRNA). Detection protocols are developed for light or electron microscopy and can be combined with immunocytochemistry to study RNA coexpression with proteins or peptides. In this chapter, we present some protocols to illustrate the diversity of in situ hybridization methods. We focus on the detection of mRNA or miRNA and show that the protocols are quite similar but use dedicated probe types, namely, oligo- or riboprobes and locked nucleic-acid probes.

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   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
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. Steinbusch, H.W., Verhofstad, A.A., and Joosten, H.W. (1978) Localization of serotonin in the central nervous system by immunohistochemistry: description of a specific and sensitive technique and some applications. Neuroscience 3, 811–819.

    Article  PubMed  CAS  Google Scholar 

  2. Liu, X., Fortin, K., and Mourelatos, Z. (2008) MicroRNAs: biogenesis and molecular functions. Brain Pathol. 18,113–121.

    Article  PubMed  CAS  Google Scholar 

  3. Landry, M., Bouali-Benazzouz, R., El Mestikawy, S., Ravassard, P., and Nagy, F. (2004) Expression of vesicular glutamate transporters in rat lumbar spinal cord, with a note on dorsal root ganglia. J. Comp. Neurol. 468, 380–394.

    Article  PubMed  CAS  Google Scholar 

  4. Landry, M., Vila-Porcile, E., Hökfelt, T., and Calas, A. (2003) Differential routing of coexisting neuropeptides in vasopressin neurons. Eur. J. Neurosci. 17, 579–589.

    Google Scholar 

  5. de Planell-Saguer, M., Rodicio, M.C., and Mourelatos, Z. (2010) Rapid in situ codetection of noncoding RNAs and proteins in cells and formalin-fixed paraffin-embedded tissue sections without protease treatment. Nat. Protoc. 5, 1061–1073.

    Article  PubMed  Google Scholar 

  6. van Gijlswijk, R.P., Zijlmans, H.J., Wiegant, J., Bobrow, M.N., Erickson, T.J., Adler, K.E., Tanke, H.J., and Raap, A.K. (1997) Fluorochrome-labeled tyramides: use in immunocytochemistry and fluorescence in situ hybridization. J. Histochem. Cytochem. 45, 375–382.

    Article  PubMed  Google Scholar 

  7. Vila-Porcile, E., Xu, Z.Q., Mailly, P., Nagy, F., Calas, A., Hökfelt, T., and Landry, M. (2009) Dendritic synthesis and release of the neuropeptide galanin: morphological evidence from studies on rat locus coeruleus neurons. J. Comp. Neurol. 516, 199–212.

    Article  PubMed  CAS  Google Scholar 

  8. Dagerlind, A., Friberg, K., Bean, A.J., and Hökfelt, T. (1992) Sensitive mRNA detection using unfixed tissue: combined radioactive and non-radioactive in situ hybridization histochemistry. Histochemistry 98, 39–49.

    Article  PubMed  CAS  Google Scholar 

  9. Kerekes, N., Landry, M., and Hökfelt, T. (1999) Leukemia inhibitory factor regulates galanin/galanin message-associated peptide expression in cultured mouse dorsal root ­ganglia; with a note on in situ hybridization methodology. Neuroscience 89, 1123–1134.

    Article  PubMed  CAS  Google Scholar 

  10. Lan, H.Y., Hutchinson, P., Tesch, G.H., Mu, W., and Atkins, R.C. (1996) A novel method of microwave treatment for detection of cytoplasmic and nuclear antigens by flow cytometry. J. Immunol. Methods. 190, 1–10.

    Article  PubMed  CAS  Google Scholar 

  11. Trembleau, A., Roche, D. and Calas, A. (1993) Combination of non-radioactive and radioactive in situ hybridization with immunohistochemistry: a new method allowing the simultaneous detection of two mRNAs and one antigen in the same brain tissue section. J. Histochem. Cytochem. 41, 489–498.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Marc Landry .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Landry, M., Salam, S.A., Moftah, M. (2011). Neuropeptide RNA Localization in Tissue Sections. In: Merighi, A. (eds) Neuropeptides. Methods in Molecular Biology, vol 789. Humana Press. https://doi.org/10.1007/978-1-61779-310-3_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-310-3_4

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-309-7

  • Online ISBN: 978-1-61779-310-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics