Cellular localization and functional expression of P-glycoprotein in rat astrocyte cultures

被引:73
作者
Ronaldson, PT
Bendayan, M
Gingras, D
Piquette-Miller, M
Bendayan, R
机构
[1] Univ Toronto, Leslie Dan Facil Fac Pharm, Dept Pharmaceut Sci, Toronto, ON M5S 2S2, Canada
[2] Univ Montreal, Fac Med, Dept Pathol & Cell Biol, Montreal, PQ H3C 3J7, Canada
关键词
astrocytes; brain parenchyma; caveolin-1; drug transport; multidrug resistance; P-glycoprotein;
D O I
10.1111/j.1471-4159.2004.02417.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We investigated the cellular/subcellular localization and functional expression of P-glycoprotein, an ATP-dependent membrane-associated efflux transporter, in astrocytes, a brain parenchyma compartment that is poorly characterized for the expression of membrane drug transporters. Analyses were carried out on primary cultures of astrocytes isolated from the cerebral cortex of neonatal Wistar rats and CTX TNA2, an immortalized rat astrocyte cell line. Both cell cultures display morphological features typical of type I astrocytes. RT-PCR analysis revealed mdr1a and mdr1b mRNA in primary cultures of astrocytes and in CTX TNA2 cells. Western blot analysis using the P-glycoprotein monoclonal C219 antibody detected a single band of appropriate size in both cell systems. Immunocytochemical analysis using the monoclonal antibodies C219 and MRK16 labeled P-glycoprotein along the plasma membrane, caveolae, coated vesicles and nuclear envelope. Immunoprecipitation studies using the caveolin-1 polyclonal H-97 antibody demonstrated that P-glycoprotein is physically associated with caveolin-1 in both cell culture systems. The accumulation of [H-3]digoxin (an established P-glycoprotein substrate) by the astrocyte cultures was significantly enhanced in the presence of standard P-glycoprotein inhibitors and an ATP depleting agent. These results demonstrate the cellular/subcellular location and functional expression of P-glycoprotein in rat astrocytes and suggest that this glial compartment may play an important role in the regulation of drug transport in the CNS.
引用
收藏
页码:788 / 800
页数:13
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