Skip to main content

Imaging In Vivo Astrocyte/Endothelial Cell Interactions at the Blood–Brain Barrier

  • Protocol
  • First Online:
Astrocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 814))

Abstract

The goal of this chapter is to highlight methods used to demonstrate in vivo changes in astrocyte expression at the blood–brain barrier (BBB). Loss of BBB integrity is seen in many acute and chronic disease conditions. However, despite the importance of the BBB to homeostasis and correct functioning of the CNS, the nature of factors responsible for the induction and maintenance of BBB properties in development and the adult remains unclear. We have studied the role of astrocytes in modulating BBB integrity in two in vivo models using a gliotoxin (3-chloropropanediol), and under hypoxic stress. 3-chloropropanediol-induced astrocytic loss within the inferior colliculus leads to loss of endothelial tight junction protein expression and loss of BBB integrity. As glial fibrillary acidic protein (GFAP)-immunopositive astrocytes repopulated the lesion, tight junction protein expression returned to paracellular domains and BBB integrity was restored. Under hypoxic conditions, increased GFAP expression was seen with changes in tight junction protein expression and loss of BBB integrity. These studies suggest a critical role for glial/endothelial interactions in regulating BBB integrity in health and disease.

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

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
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

Similar content being viewed by others

References

  1. Wolburg, H., and Lippoldt, A. (2002) Tight junctions of the blood-brain barrier: development, composition and regulation. Vasc. Pharmacol. 38, 323–337

    Article  CAS  Google Scholar 

  2. Abbott, N.J. (2002) Astrocyte-endothelial interactions and blood-brain barrier permeability. J. Anat. 200, 629-638.

    Article  PubMed  CAS  Google Scholar 

  3. Furuse, M., Hirase, T., Itoh, M., Nagafuchi, A., Yonemura, S., Tsukita, S., and Tsukita, S. (1993) Occludin—a novel integral membrane-protein localizing at tight junctions. J. Cell Biol. 123, 1777–1788.

    Article  PubMed  CAS  Google Scholar 

  4. Kniesel, U., and Wolburg, H. (2000) Tight junctions of the blood-brain barrier. Cell Mol. Neurobiol. 20, 57–76.

    Article  PubMed  CAS  Google Scholar 

  5. Plumb, J., McQuaid, S., Mirakhur, M., and Kirk, J. (2002) Abnormal endothelial tight junctions in active lesions and normal-appearing white matter in multiple sclerosis. Brain Pathol. 12, 154–169.

    Article  PubMed  Google Scholar 

  6. Abbott, N.J., Ronnback, L., Hansson, E. (2006) Astrocyte-endothelial interactions at the blood-brain barrier. Nature Rev. 7, 41–53.

    Article  CAS  Google Scholar 

  7. Rubin, L.L., Barbu, K., Bard, F., Cannon, C., Hall, D.E., Horner, H., Janatpour, M., Liaw, C., Manning, K., Morales, J., Porter, S., Tanner, L., Tomaselli, K., and Yednock, T. (1991) Differentiation of brain endothelial-cells in cell culture. Ann. NY Acad. Sci. 633, 420–425.

    Article  PubMed  CAS  Google Scholar 

  8. Sobue, K., Yamamoto, N., Yoneda, K., Hodgson, M.E., Yamashiro, K., Tsuruoka, N., Tsuda, T., Katsuya, H., Miura, Y., Asai, K., and Kato, T. (1999) Induction of blood-brain barrier properties in immortalized bovine brain endothelial cells by astrocytic factors. Neurosci. Res. 35, 155– 164.

    Article  PubMed  CAS  Google Scholar 

  9. Prat, A., Biernacki, K., Wosik, K., Antel, J.P. (2001) Glial cell influence on the human blood-brain barrier. Glia 36, 145–155.

    Article  PubMed  CAS  Google Scholar 

  10. Willis, C.L., Leach, L., Clarke, G.J., Nolan, C.C., and Ray, D.E. (2004) Reversible disruption of tight junction complexes in the rat blood-brain barrier, following transitory focal astrocyte loss. Glia 48, 1–13.

    Article  PubMed  Google Scholar 

  11. Willis, C.L., Nolan, C.C., Reith, S.N., Lister, T., Guerin, C.J., Mavroudis, G., Prior M.J.W., and Ray, D.E. (2004) Focal astrocyte loss is followed by microvascular damage, with subsequent repair of the blood-brain barrier in the apparent absence of direct astrocytic contact. Glia 45, 325–337.

    Article  PubMed  Google Scholar 

  12. Willis, C.L., Meske, D.S., and Davis, T.P. (2010) Protein kinase C activation modulates reversible increase in cortical blood-brain barrier permeability and tight junction protein expression during hypoxia and posthypoxic reoxygenation. JCBFM 30:1847–1859.

    Google Scholar 

  13. Witt, K.A., Mark, K.S., Hom, S., Davis, T.P. (2003) Effects of hypoxia-reoxygenation on rat blood-brain barrier permeability and junctional protein expression. Am. J. Physiol. Heart Circ. Physiol. 285:H2820–2831.

    PubMed  CAS  Google Scholar 

  14. Rigor, R.R., Hawkins, B.T., Miller, D.S. (2010) Activation of PKC isoform βI at the blood–brain barrier rapidly decreases P-glycoprotein activity and enhances drug delivery to the brain. JCBFM 30:1373–1383.

    Google Scholar 

Download references

Acknowledgments

The work was supported by a Medical Research Council U.K. Program grant and American Heart Association grant (AHA) SDG2170105 (CLW).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Colin L. Willis .

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

Willis, C.L. (2012). Imaging In Vivo Astrocyte/Endothelial Cell Interactions at the Blood–Brain Barrier. In: Milner, R. (eds) Astrocytes. Methods in Molecular Biology, vol 814. Humana Press. https://doi.org/10.1007/978-1-61779-452-0_34

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-452-0_34

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-451-3

  • Online ISBN: 978-1-61779-452-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics