Skip to main content

Use of In Situ Hybridization to Examine Gene Expression in the Embryonic, Neonatal, and Adult Urogenital System

  • Protocol
  • First Online:
Book cover Kidney Development

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 886))

  • 4148 Accesses

Abstract

Studies into the molecular basis of morphogenesis frequently begin with investigations into gene expression across time and cell type in that organ. One of the most anatomically informative approaches to such studies is the use of in situ hybridization, either of intact or histologically sectioned tissues. Here, we describe the optimization of this approach for use in the temporal and spatial analysis of gene expression in the urogenital system, from embryonic development to the postnatal period. The methods described are applicable for high throughput analysis of large gene sets. As such, ISH has become a powerful technique for gene expression profiling and is valuable for the validation of profiling analyses performed using other approaches such as microarrays.

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

Access this chapter

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 159.00
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

Institutional subscriptions

References

  1. Takemoto M, He L, Norlin J et al (2006) Large-scale identification of genes implicated in kidney glomerulus development and function. EMBO J 25(5):1160–1174

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Raciti D, Reggiani L, Geffers L et al (2008) Organization of the pronephric kidney revealed by large-scale gene expression mapping. Genome Biol 9(5):R84

    Article  PubMed Central  PubMed  Google Scholar 

  3. Thiagarajan RD, Georgas KM, Rumballe BA et al (2011) Identification of anchor genes during kidney development defines ontological relationships, molecular subcompartments and regulatory pathways. PLoS One 6(2):e17286

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Little MH, Brennan J, Georgas K et al (2007) A high-resolution anatomical ontology of the developing murine genitourinary tract. Gene Expr Patterns 7(6):680–699

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Chiu HS, Szucsik JC, Georgas KM et al (2010) Comparative gene expression analysis of genital tubercle development reveals a putative appendicular Wnt7 network for the epidermal differentiation. Dev Biol 344(2):1071–1087

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Georgas KM, Chiu HS, Lesieur E, Rumballe BA, Little MH (2011) Expression of metanephric nephron-patterning genes in differentiating mesonephric tubules. Dev Dyn 240(6):1600–1612

    Article  CAS  PubMed  Google Scholar 

  7. Georgas K, Rumballe B, Valerius MT et al (2009) Analysis of early nephron patterning reveals a role for distal RV proliferation in fusion to the ureteric tip via a cap mesenchyme-derived connecting segment. Dev Biol 332(2):273–286

    Article  CAS  PubMed  Google Scholar 

  8. Brunskill EW, Aronow BJ, Georgas K et al (2008) Atlas of gene expression in the developing kidney at microanatomic resolution. Dev Cell 5:781–791

    Article  Google Scholar 

  9. Rumballe BA, Georgas KM, Combes A et al (2011) Nephron formation adopts a novel spatial topology at cessation of nephrogenesis. Dev Biol 360(1):110–122

    Google Scholar 

  10. Wilkinson DG, Nieto MA (1993) Detection of messenger RNA by in situ hybridization to tissue sections and whole mounts. Methods Enzymol 225:361–373

    Article  CAS  PubMed  Google Scholar 

  11. Challen G, Gardiner B, Caruana G et al (2005) Temporal and spatial transcriptional programs in murine kidney development. Physiol Genomics 23(2):159–171

    Article  CAS  PubMed  Google Scholar 

  12. Wilhelm D, Hiramatsu R, Mizusaki H et al (2007) SOX9 regulates prostaglandin D synthase gene transcription in vivo to ensure testis development. J Biol Chem 282:10553–10560

    Article  CAS  PubMed  Google Scholar 

  13. Rumballe B, Georgas K, Little MH (2008) High-throughput paraffin section in situ hybridization and dual immunohistochemistry on mouse tissues. CSH Protoc 5030:1. doi:10.1101/pdb.prot5030

    Google Scholar 

  14. Georgas K, Rumballe B, Wilkinson L et al (2008) Use of dual section mRNA in situ hybridisation/immunohistochemistry to clarify gene expression patterns during the early stages of nephron development in the embryo and in the mature nephron of the adult mouse kidney. Histochem Cell Biol 130(5):927–942

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by NIH NIDDK grant to M.H.L. (DK070136). M.H.L. is a Principal Research Fellow of the National Health and Medical Research Council of Australia.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Melissa H. Little .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Rumballe, B.A., Chiu, H.S., Georgas, K.M., Little, M.H. (2012). Use of In Situ Hybridization to Examine Gene Expression in the Embryonic, Neonatal, and Adult Urogenital System. In: Michos, O. (eds) Kidney Development. Methods in Molecular Biology™, vol 886. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-851-1_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-851-1_20

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-850-4

  • Online ISBN: 978-1-61779-851-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics