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Functional characterization of rat plasma membrane monoamine transporter in the blood–brain and blood–cerebrospinal fluid barriers

✍ Scribed by Takashi Okura; Sayaka Kato; Yusuke Takano; Takenori Sato; Atsushi Yamashita; Riyo Morimoto; Sumio Ohtsuki; Tetsuya Terasaki; Yoshiharu Deguchi

Publisher
John Wiley and Sons
Year
2011
Tongue
English
Weight
606 KB
Volume
100
Category
Article
ISSN
0022-3549

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✦ Synopsis

This study investigated the expression and functional roles of rat plasma membrane monoamine transporter (rPMAT) in the blood-brain barrier (BBB) and the bloodcerebrospinal fluid barrier by using in vitro brain barrier model cells (TR-BBB13 and TR-CSFB3 cells) and multiple in vivo experimental techniques. Quantitative reverse transcriptionpolymerase chain reaction analysis showed relatively high expression of rPMAT mRNA in TR-BBB13 and TR-CSFB3 cells. 1-Methyl-4-phenylpyridinium (MPP + ) was transported into rPMAT-expressing cells in a sodium-independent manner. [ 3 H]MPP + was taken up concentration dependently by TR-BBB13 and TR-CSFB3 cells with K m values similar to that of rPMATexpressing cells. [ 3 H]MPP + transports into these cells were markedly inhibited by serotonin, dopamine, and cationic drugs. rPMAT small interfering RNA (siRNA) significantly suppressed the [ 3 H]MPP + uptake by TR-BBB13 cells. Intracerebrally injected [ 3 H]MPP + was eliminated from the brain parenchymal region, whereas brain [ 3 H]MPP + uptake did not increase with time during in situ brain perfusion, suggesting that the brain-to-blood transport across the BBB predominates over the blood-to-brain transport. Brain microdialysis studies revealed that the elimination across the BBB was significantly decreased by coperfusion of unlabelled MPP + , serotonin, or dopamine. [ 3 H]MPP + was also eliminated from the CSF. These findings suggest that PMAT in brain barriers functions as the brain-to-blood transporter to regulate brain concentrations of organic cations including monoamines and cationic neurotoxins.

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