Neurochemical Research

, Volume 23, Issue 12, pp 1545–1551

Preliminary Characterization of Glial-Secreted Factors Responsible for the Induction of High Electrical Resistances Across Endothelial Monolayers in a Blood-Brain Barrier Model

  • P.V. Ramsohoye
  • I.B. Fritz


Factors secreted by C6 glioma cells which induce electrical resistances across endothelial monolayers in an in vitro blood-brain barrier model have been partially characterised for the first time. These transendothelial electrical resistances (TEERs) were only evident when cell-free conditioned medium derived from C6 glioma cells was applied to the basolateral surfaces of confluent ECV304 or ECV304-9 cells which are both human umbilical vein endothelial cell lines (HUVEC). Electrical resistance values as high as 600 ohm. sq cm were obtained with this blood-brain barrier model and ultrafiltration techniques suggest that any factor(s) in the conditioned medium responsible for these TEERs have molecular masses of less than 1000 Da. Enzymic proteolysis and heat treatment carried out on the conditioned medium failed to inhibit its effect on the HUVEC monolayers suggesting that these C6 cell-secreted factors are unlikely to be proteins.

Glial blood-brain barrier 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Crone, C., and Olsen, S.-P. 1982. Electrical resistance of brain microvascular endothelium. Brain Res. 241:49-55.CrossRefPubMedGoogle Scholar
  2. 2.
    Stewart, P.A. and Wiley, M.J. 1981. Developing nervous tissue induces formation of blood-brain barrier characteristics in individual cells. Dev. Biol. 84:183-192.CrossRefPubMedGoogle Scholar
  3. 3.
    Goldstein, G.W. and Betz, A.L. 1986. The blood-brain barrier. Sci. Am. 255:70-79.CrossRefGoogle Scholar
  4. 4.
    Dermietzel, R. and Krause, D. 1991. Molecular anatomy of the blood-brain barrier as defined by immunocytochemistry. Int. Rev. Cytol. 127:57-109.CrossRefPubMedGoogle Scholar
  5. 5.
    Arthur, F.E., Shivers, R.R. and Bowman, P.D. 1987. Astrocytemediated induction of tight junctions in brain capillary endothelium: an efficient in vitro model. Dev. Brain Res. 36:155-159.CrossRefGoogle Scholar
  6. 6.
    Rubin, L.L., Hall, D.E., Porters, S., Barbu, K., Cannon, C., Horner, H.C., Janatpour, M., Liaw, C.W., Manning, K., Morales, J., Tanner, L.I., Tomaselli, K.J., and Bard, F. 1991. A cell culture model of the blood-brain barrier. J. Cell Biol. 115:1725-1735.CrossRefPubMedGoogle Scholar
  7. 7.
    Miller, D.W., Audus, K.L. and Borchardt, R.T. 1992. Application of cultured endothelial cells of the brain micro-vasculature in the study of the blood-brain barrier. J. Tiss. Cult. Meth. 14:217-224CrossRefGoogle Scholar
  8. 8.
    Raub, T.J., Kuentzel, S.L. and Sawada, G.A. 1992. Permeability of bovine brain microvessel endothelial cells in vitro: Barrier-tightening factor released from astroglioma cells. Exptal. Cell Res. 199:330-340.CrossRefGoogle Scholar
  9. 9.
    Hurst, R.D. and Fritz, I.B. 1996. Properties of an immortalised vascular endothelial/glioma cell co-culture model of the blood-brain barrier. J. Cell Physiol. 167:81-88.CrossRefPubMedGoogle Scholar
  10. 10.
    Dolman, D.E., Anderson P., Rollinson C. and Abott N.J. 1997. Characterisation of a new in vitro model of the blood-brain barrier. J. Physiol. (In press).Google Scholar
  11. 11.
    Janzer, R.C. and Raff, M.C. 1987. Astrocytes induce blood-brain barrier properties in endothelial cells. Nature. 325:253-257.CrossRefPubMedGoogle Scholar
  12. 12.
    Bradbury, M.W. 1993. The blood-brain barrier. Exptal. Physiol. 78:453-472.Google Scholar
  13. 13.
    Tao-cheng, J.H., Nagy, Z., Brightman, M.W. 1987. Tight junctions of brain endothelium in vitro are enhanced by astrocytes. J. Neurosci. 7:3293-3299.PubMedGoogle Scholar
  14. 14.
    Meyer, J., Rauh, J. and Galla, H.J. 1991. The susceptibility of cerebral endothelial cells to astroglial induction of blood-brain barrier enzymes depends on their proliferative state. J. Neurochem. 57:1971-1977.CrossRefPubMedGoogle Scholar
  15. 15.
    Tontsch, U. and Bauer, H.C. 1991. Glial cells and neurons induce blood-brain barrier related enzymes in cultured cerebral endothelial cells. Brain Res. 539:247-253.CrossRefPubMedGoogle Scholar
  16. 16.
    Dehouck, B., Dehouck, M.P., Fruchard, J.C., and Cecchelli, R. 1994. Up-regulation of the LDL receptor at the blood-brain barrier: Intercommunications between brain capillary endothelial cells and astrocytes. J. Cell Biol. 126:465-473.CrossRefPubMedGoogle Scholar
  17. 17.
    Federici, C., Camoin, L., Creminon, C., Chaverot, A.D., Strosberg, A.D., and Couraud, P.O. 1995. Cultured astrocytes release a factor that decreases endotheilin-1 secretion by brain microvessel endothelial cells. J. Neurochem. 64:1008-1015.PubMedCrossRefGoogle Scholar
  18. 18.
    Takakura, Y., Trammel, A.M., Kuentzel, S.L., Raub, T.J., Davies, A., Baldwin, S.A. and Borchardt, R.T. 1991. Biochim. Biophys. Acta. 1070:11-19.CrossRefPubMedGoogle Scholar
  19. 19.
    McMillan, M.K., Thai, L., Hong, J.S., O'Callagan, J.P. and Pennypacker, K.R. 1994. Brain injury in a dish: a model for reactive gliosis. TINS. 17:138-141.Google Scholar
  20. 20.
    Guillot, F.L. and Andus, K.L. 1991. Angiotensin peptide regulation of bovine brain microvessel endothelial cell monolayer permeability. J. Cardiovasc. Pharmacol. 18:212-218.CrossRefPubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1998

Authors and Affiliations

  • P.V. Ramsohoye
    • 1
  • I.B. Fritz
    • 1
  1. 1.Dept. of Cell PhysiologyBabraham Institute, BabrahamCambridgeUK

Personalised recommendations