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Functional expression of the P-glycoproteinmdr in primary cultures of bovine cerebral capillary endothelial cells

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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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