Skip to main content
SpringerLink
Log in

In Situ Hybridization with 35S-Labeled Probes for Retinoid Receptors

  • Protocol
Retinoid Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 89))

Abstract

In situ hybridization (ISH) is a technique that enables one to localize an endogenous mRNA in histologtcal and cytological samples to the resolution of nearly single cells. For this reason, ISH is widely used in the fields of genetics, developmental biology, and neurobiology, as well as for dtagnostic purposes in medicine. Our laboratory has established conditions for ISH using 35S-labeled antisense-RNA probes (riboprobes), whtch allow the specific detection of each type of murine retinoic acid receptor (RAR), retinoid X receptor (RXR), cellular retinol-binding protein (CRBP), and cellular retinotc acid binding protein (CRABP) gene transcripts. ISH studies have thus revealed that most of the retinoid receptor genes have complex and distinct expression patterns during mouse development (1–6). The protocol described hereafter has been inspired by the pioneermg work of Angerer and colleagues (7). It has been designed for histological sections of mouse embryos or adult ttssues, but can be applied to samples from other species, including human. Nevertheless, slightly distinct ISH protocols have been described for the detection of RAR transcripts in chick (8, 9) and Xenopus embryos (10).

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Dollé, P., Ruberte, E, Kastner, P., Petkovtch, M., Stoner, C M., Gudas, L. J, and Chambon, P (1989) Differential expression of genes encoding α, β and γ retinoic acid receptors and CRABP in the developmg limbs of the mouse Nature 262, 702–705.

    Article  Google Scholar 

  2. Dollé, P, Ruberte, E, Leroy, P, Morriss-Kay, G, and Chambon, P (1990) Retinotc acid receptors and cellular retinoid binding proteins I A Systematic study of their differential pattern of transcription during mouse organogenests Development 110, 1133–1151

    PubMed  Google Scholar 

  3. Ruberte, E., Dollè, P., Krust, A, Zelent, A, Morriss-Kay, G, and Chambon, P (1990) Specific spatial and temporal dtstrtbution of retinoic acid receptors gamma transcripts during mouse embryogenests Development 108, 213–222

    PubMed  CAS  Google Scholar 

  4. Ruberte, E, Dollé, P., Chambon, P., and Momss-Kay, G (1991) Retinoic acid receptors and cellular retinoid binding proteins II Their differential pattern of tran-scription dunng early morphogenesis in mouse embryos Development 111, 45–60.

    PubMed  CAS  Google Scholar 

  5. Ruberte, E, Fnedench, V., Chambon, P., and Morriss-Kay, G (1993) Retinoic acid receptors and cellular retinoid binding proteins. III Then differential transcript distri-bution during mouse nervous system development Development 118, 267–282

    PubMed  CAS  Google Scholar 

  6. Dollé, P, Fraulob, V, Kastner, P., and Chambon, P (1994) Developmental expression of murme retinoid X receptor (RXR) genes Mech Dev. 45, 91–104.

    Article  PubMed  Google Scholar 

  7. Cox, K H, DeLeon, D. V, Angerer, L. M, and Angerer, R. C (1984) Detection of mRNAs in sea urchin embryos by in situ hybridization using asymtnetric RNA probes. Dev Biol 101, 485–502.

    Article  PubMed  CAS  Google Scholar 

  8. Wedden, S., Pang, K., and Eichele, G. (1989) Expresston pattern of homeobox containing genes durmg chick embryogenests Development 105, 639–650

    PubMed  CAS  Google Scholar 

  9. Smith, S M., and Eichele, G. (1991) Temporal and regional differences in the expression pattern of distinct retinoic acid receptor-beta transcripts in the chick embryo Development 111, 245–252

    PubMed  CAS  Google Scholar 

  10. Ellmger-Ziegelbauer, H and Dreyer, C (1991) A retinoic acid receptor expressed in the early development of Xenopus laevis Genes Dev 5, 94–104

    Article  Google Scholar 

  11. Polak, J. M and McGee, J O’D (eds.) (1990) In Situ Hybridzzation. Principles and Practice Oxford University Press, Oxford, UK

    Google Scholar 

  12. Wilkinson, D G (ed ) (1992) In Situ Hybridization. A Practical Approach IRL, Oxford University Press.

    Google Scholar 

  13. Prophet, E B, Mills, B, Arrington, J. B, and Sobin, L H. S (eds.) (1992) Laboratory Methods in Histotechnology. American Registry of Pathology, Washington DC

    Google Scholar 

  14. Wilkinson, D G., and Green, J (1992) In situ hybridization and the three-dimensional reconstruction of serial sections, in Postlmplantatlon Manimalian Embryos. A Practical Approach (Copp, A J., and Cockroft, D. L, eds.) Oxford University Press, pp 155–171

    Google Scholar 

  15. Monaghan, A P, Davidson, D R, Sime, C., Graham, E, Baldock, R., Bhattacharya, S. S., and Hill, R. E (1991) The Msh-like homeobox gene define domains in the developmg vertebrate eye. Development 112, 1053–1061

    PubMed  CAS  Google Scholar 

  16. Vonesch, J L, Nakshatri, H, Philippe, M., Chambon, P, and Dollé, P (1994) Stage and tissue-specific expression of the alcohol dehydrogenase 1 (Adh-1) gene during mouse development. Dev. Dynamics 199, 199–213

    Article  CAS  Google Scholar 

  17. Smith, S. M (1994) Retinoic acid receptor isoform β2 is an early marker for alimentary tract and central nervous system positional specification in the chicken Dev. Dynamics 200, 14–25

    Article  CAS  Google Scholar 

  18. Smith, S M, Kirstein, I J., Wang, Z S, Fallon, J. F, Kelley, J., and Bradshaw-Rouse, J (1995) Differential expression of retinoic acid receptor-β tsoforms during chick limb ontogeny Dev Dynamics 202, 54–66 22

    Article  CAS  Google Scholar 

  19. Pfeffer, P L and De Robertts, E M. (1994) Regional specificity of RARγ isoforms in Xenopus development Mech. Dev 45, 147–153

    Article  PubMed  CAS  Google Scholar 

  20. Sambrook, J., Fritsch, E F., and Maniatis, T (1989) Molecular Cloning—A Laboratory Manual, 2nd ed, Cold Spring Harbor Laboratory, Cold spring Harbor, NY.

    Google Scholar 

  21. Ausubel, F M., Brent, R., Kingston, R. E., Moore, D. D, Setdman, J G, Smith, J. A., and Struhl, K. (eds.) (1987) Current Protocols in Molecular Biology. Green Publishing Associates and Wiley Interscience, Wiley & Sons.

    Google Scholar 

  22. Décimo, D., Georges-Labouesse, E., and Dollé, P. (1995) In situ hybridization of nucleic acid probes to cellular RNA, in Gene Probes 2, (Hames, B. D., and Higgms, S., eds.), Oxford Umversity Press, pp. 183–210

    Google Scholar 

  23. Noji, S, Yamaai, T., Koyama, E., Nohno, T, Fujimoto, W., Arata, J., and Tamguchi, S. (1989) Expression of retinoic acid receptor genes in keratintzing front of skin. FEBS 259, 86–90.

    Article  CAS  Google Scholar 

  24. Finzi, E, Blake, M. J., Celano, P., Skouge, J., and Diwan, R. (1992) Cellular localization of retinoic acid receptor-gamma expression in normal and neoplastic skin Am. J. Pathool 140, 1463–1471.

    CAS  Google Scholar 

  25. Xu, X C, Clifford, J. L, Hong, W. K, and Lotan, R. (1994) Detection of nuclear retinoic acid receptor mRNAs in histological tissue sections using non-radioactive in situ hybridizatton htstochemistry. Diagnostic Mol. Pathol 3, 122–131

    Article  CAS  Google Scholar 

  26. Xu, X. C., Ro, J Y, Lee, J S, Shin, D. M, Hong, W K., and Lotan, R. (1994) Differential expression of nuclear retinoic acid receptors in normal, premahgnant, and malignant head and neck tissues. Cancer Res 54, 3580–3587

    PubMed  CAS  Google Scholar 

  27. Mangelsdorf, D J, Borgmeyer, U., Heyman, R A., Zhou, J Y., Ong, E S., Oro, A. E, Kaktzuka, A, and Evans, R. (1992) Characterization of three RXR genes that mediate the action of 9-cis retinoic acid. Genes Dev. 6, 329–344

    Article  PubMed  CAS  Google Scholar 

  28. Rowe, A., Richman, J M., and Brtckell, P. M (1991) Retinoic acid treatment alters the distrtbutton of retinotc acid receptor-β transcripts in the embryonic chick face. Development 111, 1007–1016.

    PubMed  CAS  Google Scholar 

  29. Michaille, J. J., Blanche, S., Kanzler, B., Garnier, J M, and Dhouailly, D. (1994) Characterization of cDNAs encoding the chick retinoic acid receptor γ2 and preferential distribution of retmic acid receptor γ transcripts durmg chick skin development. Dev. Dynamics 201, 334–343

    Article  CAS  Google Scholar 

  30. Rowe, A., Eager, N. S. C., and Brickell, P. (1991) a member of the RXR nuclear receptor family is expressed in neural-crest-dertved cells of the developing chick peripheral nervous system. Development 111, 771–778

    PubMed  CAS  Google Scholar 

  31. White, J A., Boffa, M B., Jones, B, and Petkovich, M. (1994) a zebrafish retinoic acid receptor expressed in the regenerating caudal fin. Development 120, 1861–1872

    PubMed  CAS  Google Scholar 

  32. Blumberg, B., Mangelsdorf, D J., Dyck, J. A, Bittner, D A, Evans, R M, and De Robertis, E M (1992) Multiple retinoid-responsive receptors in a single cell families of retinoid “X” receptors and retinoic acid receptors in the Xenopus egg. Proc Natl. Acad. Sci USA 89, 2321–2325.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGMBC), CNRS/INSERM/ULP/Collége de France, CU de Strasbourg, France

    Karen Niederreither & Pascal Dollé

Authors
  1. Karen Niederreither
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pascal Dollé
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. University of Newcastle Upon Tyne, UK

    Christopher P. F. Redfern

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc, Totowa, NJ

About this protocol

Cite this protocol

Niederreither, K., Dollé, P. (1998). In Situ Hybridization with 35S-Labeled Probes for Retinoid Receptors. In: Redfern, C.P.F. (eds) Retinoid Protocols. Methods in Molecular Biology, vol 89. Humana Press. https://doi.org/10.1385/0-89603-438-0:247

Download citation

  • DOI: https://doi.org/10.1385/0-89603-438-0:247

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-438-9

  • Online ISBN: 978-1-59259-573-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation