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The Effect of Conditioned Medium of Porcine Astrocytes and Different Cell Lines on the Activity of the Marker Enzymes Alp and γ-GT in Endothelial Cells of Porcine Brain Microvessels

  • Barbara Ahlemeyer
  • Sabine Matys
  • Peter Brust
Part of the Advances in Behavioral Biology book series (ABBI, volume 46)

Summary

The conditioned media of subconfluent and confluent monolayers of porcine astrocytes and different cell lines were used to reinduce in primary cultured endothelial cells of porcine brain microvessels in vivo properties like high activities of the marker enzymes alkaline phosphatase (ALP) and γ-glutamyltranspeptidase (γ-GT). After 7 days in culture, the activity of ALP increased only by incubation of cerebral endothelial cells with conditioned media from confluent adult porcine astrocytes and CHO-cells (fibroblasts), whereas the activity of γ-GT was significantly increased by conditioned media of astrocytes, CHOcells, chondrocytes and adenocarcinoma cells. However, the increase in γ-GT seems to be the result of an increase in the number of cells on the top of the monolayer with high γ-GT activities as revealed by histochemistry. The mechanism of the inductive effect on γ-GT seems to be not astrocyte specific and has to be clarified.

Keywords

Conditioned Medium Porcine Brain Cerebral Endothelial Cell Brain Microvessel Endothelial Cell Adult Porcine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Résumé

Des milieux de culture conditionnés par des monocouches confluentes et subconfluentes d'astrocytes de porc et de differentes lignées cellulaires ont été utilisées pour réinduire, dans des cultures primaires de microvaisseaux de porc, des propriétés de la barrière hémato-encéphalique in vivo telles les fortes activités enzymatiques de la phosphatase alcaline (PA) ou de la γ-glutamytranspeptidase (γ-GT). Après sept jours de culture, l’activité de la PA a pu être réinduite seulement par les milieux conditionnés par les cultures confluentes d’astrocytes de porc adulte et d’une lignée de fibroblastes (CHO). Une augmentation de l’activité de la γ-GT a pu, dans les mêmes conditions, être observée avec des milieux conditionnés d’astrocytes, de cellules CHO, de chondrocytes et de cellules d’adénocarcinome. Toutefois, l’augmentation de l’activité de la γ-GT semble être le resultat de la multiplication au dessus de la monocouche de cellules possédant une forte activité γ-GT, comme le montrent des expériences d’histochimie. L’effet inducteur des milieux de cultures sur l’activité de la γ-GT ne semble pas spécifique des astrocytes et reste à élucider.

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References

  1. 1.
    Joó, F., The cerebral microvessels in culture, an update, J. Neurochem. 58: 1, 1992.PubMedCrossRefGoogle Scholar
  2. 2.
    Fukushima, H., Fujimoto, M., and Ide, M., Quantitative detection of blood-brain barrier-associated enzymes in cultured endothelial cells of porcine brain microvessels, In Vitro Cell. Del). Biol. 26: 612, 1990.CrossRefGoogle Scholar
  3. 3.
    Meyer, J., Mischek, U., Veyhl, M., Henze], K., and Galla, H.J., Blood-brain barrier characteristics enzymatic properties in cultured brain capillary endothelial cells, Brain Res. 514: 305, 1990.PubMedCrossRefGoogle Scholar
  4. 4.
    DeBault, L.E., and Cancilla P.A., y-Glutamyltranspeptidase in isolated brain endothelial cells: induction by glial cells in vitro, Science 207: 653, 1980.PubMedCrossRefGoogle Scholar
  5. 5.
    Tao-Cheng, J.H., Nagy, Z., and Brightman, M.W., Tight juctions of brain endothelium in vitro are enhanced by astroglia, J. Neurosci. 7: 3293, 1987.PubMedGoogle Scholar
  6. 6.
    Laterra, J., Guerin, C., and Goldstein, G.W., Astrocytes induce neural microvascular endothelial cells to form capillary-like structures in vitro, J. Cell. Physiol. 144: 204, 1990.PubMedCrossRefGoogle Scholar
  7. 7.
    Meyer, J., Rauh, J., and Galla, H.J., The susceptibility of cerebral endothelial cells to astroglial induction of blood-brain barrier enzymes depends on their proliferative state, J. Neurochem. 57:1971, 1991.Google Scholar
  8. 8.
    Tontsch, U., and Bauer, H.C., Glial cells and neurons induce blood-brain barrier related enzymes in cultured cerebral endothelial cells, Brain Res. 539: 247, 1991.PubMedCrossRefGoogle Scholar
  9. 9.
    Hurwitz, A.A., Berman, J.W., Rashbaum, W.K., and Lyman, W.D., Human fetal astrocytes induce the expression of blood brain barrier specific proteins by autologous endothelial cells, Brain Res. 625: 238, 1993.PubMedCrossRefGoogle Scholar
  10. 10.
    Arthur, F.E., Shivers, R.R., and Bowman P.D., Astrocyte-mediated induction of tight junctions in brain capillary endothelium: an efficient in vitro model, Dev. Brain Res. 36: 155, 1987.CrossRefGoogle Scholar
  11. 11.
    Maxwell, K., Berliner, J.A., and Cancilla, P.A., Induction ofy-glutamyltranspeptidase in cultured cerebral endothelial cells by a product released by astrocytes, Brain Res. 410: 309, 1987.PubMedCrossRefGoogle Scholar
  12. 12.
    Rubin, L.L., Hall, D.E., Porter, 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., A culture model of the blood-brain barrier, J. Cell Biol. 115: 1725, 1991.PubMedCrossRefGoogle Scholar
  13. 13.
    Raub, T.J., Kuentzel, S.L., and Sawada, G.A., Permeability of bovine brain microvessel endothelial cells in vitro: barrier tightening by a factor released from astroglioma cells, Exp. Cell. Res. 199: 330, 1992.PubMedCrossRefGoogle Scholar
  14. 14.
    Wolburg, H., Neuhaus, J., Kniesel, U., Krauß, B., Schmid, E.M., Ocalan, M., Farrell, C., and Risau, W., Modulation of tight junction structure in blood-brain barrier endothelial cells, J Cell Sci. 107: 1347, 1994.PubMedGoogle Scholar
  15. 15.
    Takemoto, H., Kaneda, K., Hosokawa, M., Ide, M., and Fukushima, H., Conditioned media of glial cell lines induce alkaline phosphatase activity in cultured artery endothelial cells, FEBSLett_ 350: 99, 1994.CrossRefGoogle Scholar
  16. 16.
    Mizuguchi, H., Hashioka, Y., Fujii,A., Utoguchi, N., Kubo, K., Nakagawa, S., Baba, A., and Mayumi, T., G1ial extracellular matrix modulates y-glutamyltranspeptidase activity in cultured bovine brain capillary and bovine aortic endothelial cells, Brain Res. 651: 155, 1994.PubMedCrossRefGoogle Scholar
  17. 17.
    Mischek, U., Meyer, J., and Galla, H.J., Characterization of y-glutamyl transpeptidase activity of cultured endothelial cells from porcine brain capillaries, Cell. Tissue Res. 256: 221, 1989.Google Scholar
  18. 18.
    Meyer J., Mischek, U., Vehyl, M., Henzel, K., and Galla, H.J., Blood-brain barrier characteristic enzymatic properties in cultured brain capillary endothelial cells. Cell Tissue Res. 256: 221, 1989.PubMedCrossRefGoogle Scholar
  19. 19.
    Roux, F., Durieu-Trautmann O., Chaverot, N., Claire, M., Mailly, P., Bourre, J.M., Strosberg, A.D., and Couraud, P.O., Regulation of gamma-glutamyltranspeptidase and alkaline phosphatase activities in immortalized rat brain microvessel endothelial cells, J Cell. Physiol. 159: 101. 1994.PubMedCrossRefGoogle Scholar
  20. 20.
    Wang. B.L., Grammas, P.. and DeBault, L.E., Characterization of a y-glutamyltranspeptidase positive subpopulation of endothelial cells in a spontaneous tube-forming clone of rat cerebral resistance-vessel endothelium, J. Cell. Phvsiol. 156: 531, 1993.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • Barbara Ahlemeyer
    • 1
  • Sabine Matys
    • 1
  • Peter Brust
    • 1
  1. 1.Research Center RossendorfInstitute of Bioinorganic and Radiopharmaceutic ChemistryDresdenGermany

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