title: Functional test of multidrug transporter activity in hippocampal-neocortical brain slices from epileptic patients.
abstract: About 70% of the patients suffering from temporal lobe epilepsy (TLE) are resistant to currently available antiepileptic drugs (AEDs). For them one therapeutic option to achieve seizure control is to undergo epilepsy surgery. Expression of multidrug transporters is upregulated in resected tissue specimens from TLE patients, as well as in animal models of chronic epilepsy, which might lead to altered tissue availability of AEDs and therefore contribute to drug refractoriness. Here we describe a functional test of multidrug transporter activity in brain slices from TLE patients based on intracellular accumulation of the fluorescent multidrug transporter substrate calcein and compare functional data to the expression pattern of multidrug transporters. The rate of cytosolic calcein fluorescence increase was altered by inhibitors of multidrug transport such as probenecid (400 μM) and verapamil (40 μM) in a subset of slices, indicating the presence of functional multidrug transport proteins in human epileptic tissue. Interestingly, there were differences between the expression pattern of multidrug transporters and their ability to remove calcein-AM. Consequently, in vitro studies on multidrug transporters should always include functional tests of their activity as expression alone is not necessarily conclusive.
authors.affiliation: Institute for Neurophysiology, Charité-Universitätsmedizin Berlin, 13347 Berlin, Germany. richard.kovacs@charite.de
authors.forename: Richard
authors.initials: R
authors.lastname: Kovács
authors.forename: Claudia
authors.initials: C
authors.lastname: Raue
authors.forename: Siegrun
authors.initials: S
authors.lastname: Gabriel
authors.forename: Uwe
authors.initials: U
authors.lastname: Heinemann
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000277:Adjuvants, Pharmaceutic / Q000494:pharmacology; D000927:Anticonvulsants / Q000494:pharmacology; D004827:Epilepsy / Q000473:pathology*; D005452:Fluoresceins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D006801:Humans; D066298:In Vitro Techniques; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism*; D019579:Neocortex / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011339:Probenecid / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000277:Adjuvants, Pharmaceutic; D000927:Anticonvulsants; D005452:Fluoresceins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D009419:Nerve Tissue Proteins; D011339:Probenecid; D014700:Verapamil; C085925:calcein AM; C007740:fluorexon