Skip to main content

Localization and Distribution of Cyclooxygenase-2 in Brain Tissue by Immunohistochemistry

  • Protocol

Part of the book series: Methods in Molecular Biology ((MIMB,volume 120))

Abstract

Prostaglandin production is dependent on the presence of an enzyme called prostaglandin G/H synthase, better known as cyclooxygenase, that catalyzes the first steps in the conversion from arachidonic acid. A cDNA for cyclooxygenase was first cloned from sheep seminal vesicles in 1988 (1,2). In 1991, we and others reported the cloning of a second isoform of cyclooxygenase, now known as COX-2, that is rapidly induced in response to growth factors and proinflammatory cytokines and downregulated by glucocorticoid hormones (36).

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   109.99
Price excludes VAT (USA)
  • Compact, lightweight 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. Merlie, J. P., Fagan, D., Mudd, J., and Needlman, P. (1988) Isolation and characterization of the complementary DNA for sheep seminal vesicle prostaglandin endoperoxide synthase (cyclooxygenase). J. Biol. Chem. 263, 3550–3553.

    PubMed  CAS  Google Scholar 

  2. DeWitt, D. L. and Smith, W. L. (1988) Primary structure of prostaglandin G/H synthase from sheep vesicular gland determined from the complementary DNA sequence. Proc. Natl. Acad. Sci. USA 85, 1412–1416.

    Article  PubMed  CAS  Google Scholar 

  3. Kujubu, D. A., Fletcher, B. S., Varnum, B. C., Lim, R. W., and Herschmann, H. R. (1991) TIS10, a phorbol ester tumor promoter-inducible mRNA from Swiss 3T3 cells, encodes a novel prostaglandin synthase/cyclooxygenase homologue. J. Biol. Chem. 266, 12,866–12,872.

    PubMed  CAS  Google Scholar 

  4. O’Banion, M. K., Sadowski, H. B., Winn, V., and Young, D. A. (1991) A serumand glucocorticoid-regulated 4 kb mRNA encodes a cyclooxygenase related protein. J. Biol. Chem. 266, 23,261–23,267.

    PubMed  Google Scholar 

  5. Xie, W., Chipman, J. G., Robertson, D. L., Erikson, R. L., and Simmons, D. L. (1991) Expression of a mitogen-responsive gene encoding prostaglandin synthase is regulated by mRNA splicing. Proc. Natl. Acad. Sci. USA 88, 2692–2696.

    Article  PubMed  CAS  Google Scholar 

  6. O’Banion, M. K., Winn, V. D., and Young, D. A. (1992) cDNA cloning and functional activity of a glucocorticoid-regulated inflammatory cyclooxygenase. Proc. Natl. Acad. Sci. USA 89, 4888–4892.

    Article  PubMed  Google Scholar 

  7. Smith, W. L., Garavito, R. M., and DeWitt, D. L. (1996) Prostaglandin endoperoxidase H synthases (cyclooxygenases) −1 and −2. J. Biol. Chem. 271, 33,157–33,160.

    Article  PubMed  CAS  Google Scholar 

  8. Williams, C. S. and DuBois, R. N. (1996) Prostaglandin endoperoxidase synthase: why two isoforms? Am. J. Physiol. 270, G393–400.

    PubMed  CAS  Google Scholar 

  9. Harris, R. C., McKanna, J. A., Akai, Y., Jacobson, H. R., DuBois, R. N., and Breyer, M. D. (1994) Cyclooxygenase-2 is associated with the macula densa of rat kidney and increases with salt restriction. J. Clin. Invest. 94, 2504–2510.

    Article  PubMed  CAS  Google Scholar 

  10. Breder, C. D., DeWitt, D., and Kraig, R. P. (1995) Characterization of inducible cyclooxygenase in rat brain. J. Comp. Neurol. 355, 296–315.

    Article  PubMed  CAS  Google Scholar 

  11. Yamagata, K., Andreasson, K. I., Kaufmann, W. I., Barnes, C. A., and Worley, P. F. (1993) Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 11, 371–386.

    Article  PubMed  CAS  Google Scholar 

  12. Kaufmann, W. E., Worley, P. F., Pegg, J., Bremer, M., and Isakson, P. (1996) Cox-2, a synaptically-induced enzyme, is expressed by excitatory neurons at postsynaptic sites in rat cerebral cortex. Proc. Natl. Acad. Sci. USA 93, 2317–2321.

    Article  PubMed  CAS  Google Scholar 

  13. O’Banion, M. K., Dusel, J. C., Chang, J. W., Kaplan, M. D., and Coleman, P. D. (1996) Interleukin-1β induces prostaglandin G/H synthase-2 (cyclooxygenase-2) in primary murine astrocyte cultures. J. Neurochem. 66, 2532–2540.

    Article  PubMed  Google Scholar 

  14. Thore, C. R., Nam, M. J., and Busija, D. W. (1996) Immunofluorescent localization of constitutive and inducible prostaglandin H synthase in ovine astroglia. J. Comp. Neurol. 367, 1–9.

    Article  PubMed  CAS  Google Scholar 

  15. Minghetti, L., Polazzi, E., Nicolini, A., Créminon, C., and Levi, G. (1996) Interferon gamma and nitric oxide down-regulate lipopolysaccharide-induced prostanoid production in cultured rat microglial cells by inhibiting cyclooxygenase-2 expression. J. Neurochem. 66, 1963–1970.

    Article  PubMed  CAS  Google Scholar 

  16. Bauer, M. K. A., Lieb, K., Schulzeostohoff, K., Berger, M., Gebicke-Haerter, P. J., Bauer, J., and Fiebich, B. L. (1997) Expression and regulation of cyclo-oxygenase-2 in rat microglia. Eur. J. Biochem. 243, 726–731.

    Article  PubMed  CAS  Google Scholar 

  17. Breder, C. D. and Saper, C. B. (1996) Expression of inducible cyclooxygenase mRNA in the mouse brain after systemic administration of bacterial lipopolysac-charide. Brain Res. 713, 64–69.

    Article  PubMed  CAS  Google Scholar 

  18. Breitner, J. C. S. (1996) Inflammatory processes and antiinflammatory drugs in Alzheimer’s disease: a current appraisal. Neurobiol. Aging 17, 789–794.

    Article  PubMed  CAS  Google Scholar 

  19. Rich, J. B., Rasmusson, D. X., Folstein, M. F., Carson, K. A., Kawas, C., and Brandt, J. (1995) Nonsteroidal anti-inflammatory drugs in Alzheimer’s disease. Neurology 45, 51–55.

    PubMed  CAS  Google Scholar 

  20. Stewart, W. F., Kawas, C., Corrada, M., and Metter, E. J. (1997) Risk of Alzheimer’s disease and duration of NSAID use. Neurology 48, 626–632.

    PubMed  CAS  Google Scholar 

  21. Nogawa, S., Zhang, F., Ross, M. E., and Iadecola, C. (1997) Cyclooxygenase-2 gene expression in neurons contributes to ischemic brain damage. J. Neurosci. 17, 2746–2755.

    PubMed  CAS  Google Scholar 

  22. McKanna, J. A. and Zhang, M. Z. (1997) Immunohistochemical localization of lipocortin 1 in rat brain is sensitive to pH, freezing, and dehydration. J. Histochem. Cytochem. 45, 527–538.

    PubMed  CAS  Google Scholar 

  23. Watson, R. E., Jr., Wiegnad, S. J., Clough, R. W., and Hoffmann, G. E. (1986) Use of cryoprotectant to maintain long-term peptide immunoreactivity and tissue morphology. Peptides 7, 155–159.

    Article  PubMed  CAS  Google Scholar 

  24. Pritchard, K. A., Jr., O’Banion, M. K., Miano, J. M., Vlasic, N., Bhatia, U. G., Young, D. A., and Stemerman, M. B. (1994) Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. J. Biol. Chem. 269, 8504–8509.

    PubMed  CAS  Google Scholar 

Download references

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Humana Press Inc.

About this protocol

Cite this protocol

Kerry O’Banion, M., Olschowka, J.A. (1999). Localization and Distribution of Cyclooxygenase-2 in Brain Tissue by Immunohistochemistry. In: Lianos, E.A. (eds) Eicosanoid Protocols. Methods in Molecular Biology, vol 120. Humana Press. https://doi.org/10.1385/1-59259-263-5:55

Download citation

  • DOI: https://doi.org/10.1385/1-59259-263-5:55

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-667-3

  • Online ISBN: 978-1-59259-263-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics