Abstract
The P-glycoproteinmdr is expressed not only in tumoral cells, but also in nontransformed cells, including the specialized endothelial cells of brain capillaries which build up the blood-brain barrier. Since all previously identified blood-brain barrier markers are rapidly lost when cerebral capillary endothelial cells are maintained in primary culture, we have investigated whether P-glycoprotein (P-gp) would follow the same rule, in order to address the influence of the cerebral environment on the specific P-gp expression in the brain endothelium. As compared to freshly isolated purified cerebral capillaries, P-glycoprotein was detected by immunochemistry at a high level in 5–7 day primary cultures. In our culture conditions, P-glycoprotein was immunodetected at a lower molecular weight than that found in freshly isolated capillaries. Enzymatic deglycosylation led to the same 130 kDa protein for both fresh and cultured samples, suggesting that P-gp post-translational modifications were altered in primary cultures. However, studies on the uptake and efflux of the P-gp substrate [3H]vinblastine, and on the effect of variousmdr reversing agents on the uptake and efflux, clearly indicated that the efflux pump function of the P-glycoprotein was maintained in primary cultures of bovine cerebral capillary endothelial cells. P-Glycoprotein may thus represent the first blood-brain barrier marker which is maintained in cerebral endothelial cells cultured in the absence of factors originating from the brain parenchyma.
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Abbreviations
- BBB:
-
blood-brain barrier
- BCEC:
-
brain capillary endothelial cells
- γ-GT:
-
γ-glutamyltranspeptidase
- HBSS:
-
Hank's balanced salt solution
- Mab:
-
monoclonal antibody
- mdr :
-
multidrug resistance
- P-gp:
-
P-glycoprotein
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Lechardeur, D., Scherman, D. Functional expression of the P-glycoproteinmdr in primary cultures of bovine cerebral capillary endothelial cells. Cell Biol Toxicol 11, 283–293 (1995). https://doi.org/10.1007/BF00757626
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DOI: https://doi.org/10.1007/BF00757626