Expression and Functional Characterization of Rat Organic Anion Transporter 3 (rOat3) in the Choroid Plexus

Nagata, Yoshinori; Kusuhara, Hiroyuki; Endou, Hitoshi; Sugiyama, Yuichi

doi:10.1124/mol.61.5.982

Cited by 138 publications

(132 citation statements)

References 30 publications

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“…Again, this finding is consistent with the reversal of functional polarity between kidney and choroid plexus as discussed previously. Finally, localization of Oat3 on the apical membrane in choroid plexus (and the basolateral membrane of brain capillaries) was confirmed by immunostaining [70,71].…”

Section: Oat2 (Slc22a7)mentioning

confidence: 85%

Organic anion transporters: discovery, pharmacology, regulation and roles in pathophysiology

VanWert

Gionfriddo

Sweet

2009

Biopharm & Drug Disp

222

238

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Our understanding of the mechanisms behind inter-and intra-patient variability in drug response is inadequate. Advances in the cytochrome P450 drug metabolizing enzyme field have been remarkable, but those in the drug transporter field have trailed behind. Currently, however, interest in carrier-mediated disposition of pharmacotherapeutics is on a substantial uprise. This is exemplified by the 2006 FDA guidance statement directed to the pharmaceutical industry. The guidance recommended that industry ascertain whether novel drug entities interact with transporters. This suggestion likely stems from the observation that several novel cloned transporters contribute significantly to the disposition of various approved drugs. Many drugs bear anionic functional groups, and thus interact with organic anion transporters (OATs). Collectively, these transporters are nearly ubiquitously expressed in barrier epithelia. Moreover, several reports indicate that OATs are subject to diverse forms of regulation, much like drug metabolizing enzymes and receptors. Thus, critical to furthering our understanding of patient-and condition-specific responses to pharmacotherapy is the complete characterization of OAT interactions with drugs and regulatory factors. This review provides the reader with a comprehensive account of the function and substrate profile of cloned OATs. In addition, a major focus of this review is on the regulation of OATs including the impact of transcriptional and epigenetic factors, phosphorylation, hormones and gender.

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Section: Oat2 (Slc22a7)mentioning

confidence: 85%

Organic anion transporters: discovery, pharmacology, regulation and roles in pathophysiology

VanWert

Gionfriddo

Sweet

2009

Biopharm & Drug Disp

222

238

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“…The expression of Zn-transporter-1 (ZnT1) in rat choroidal epithelium has been verified by immunohistochemistry (Wang et al, 2004), which regulates intracellular Zn 2+ by acting as a zinc ion efflux pump (Sekler et al, 2002). The CP also possesses the transporters responsible for transport of organic chemicals, such as multi-drug resistance protein-1 (MRP1) (Rao et al, 1999;Wijnholds et al, 2000), organic anion transporter (Oat3) (Nagata et al, 2002), and organic cation transporter-1 (OCT1) (Sweet et al, 2001). The current study clearly demonstrated the presence of all these transporters in Z310 cells.…”

Section: Discussionmentioning

confidence: 99%

Use of Z310 cells as an in vitro blood–cerebrospinal fluid barrier model: Tight junction proteins and transport properties

Shi

Wang

et al. 2008

Toxicology in Vitro

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Immortalized rat choroidal epithelial Z310 cells have the potential to become an in vitro model for studying transport of materials at blood-cerebrospinal fluid barrier (BCB) (Shi and Zheng, Brain Research 1057:37-48, 2005). This study was designed to demonstrate the presence of tight junction properties in Z310 cells and the functionality of Z310 monolayer in transport of selected model compounds. Western blot analyses revealed the presence of claudin-1, ZO-1, and occludin in Z310 cells. Transmission electron microscopy showed a "tight junction" type of structure in the sub-apical lateral membranes between adjacent Z310 cells. Real-time RT-PCR revealed that Z310 cells expressed representative transporters such as DMT1, MTP1, TfR, p-glycoprotein, ATP7A, ZnT1, ABCC1, Oat3, OCT1 and OB-Ra. Moreover, Z310 cells cultured in a two-chamber Transwell device possessed the ability to transport zidovudine (anionic drug), thyroxine (hormone), thymidine (nucleoside), and leptin (large polypeptide) with kinetic properties similar to those obtained from the in vitro model based on primary culture of choroidal epithelial cells. Taken together, these data indicate that the Z310 BCB model expresses major tight junction proteins and forms a tight barrier in vitro. The model also exhibits the ability to transport substances of various categories across the barrier.

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“…rOat3 accepts amphipathic organic anions such as E 2 17␤G and E 1 S as well as PAH as its substrates (Kusuhara et al, 1999;Sugiyama et al, 2001). In the brain, rOat3 has been shown to be expressed in the choroid plexus (Nagata et al, 2002) as well as in the brain capillaries (Ohtsuki et al, 2002). In the choroid plexus, rOat3 is responsible for the uptake of PAH and PCG from the cerebrospinal fluid (Nagata et al, 2002).…”

mentioning

confidence: 99%

Contribution of Organic Anion Transporter 3 (Slc22a8) to the Elimination of p-Aminohippuric Acid and Benzylpenicillin across the Blood-Brain Barrier

Kikuchi

Kusuhara²,

Sugiyama³

et al. 2003

The Journal of Pharmacology and Experimental Therapeutics

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104

118

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The role of rat organic anion transporter 3 (rOat3; Slc22a8) in the efflux transport at the blood-brain barrier (BBB) was characterized. The expression of rOat1, rOat2, and rOat3 in the brain capillary endothelial cells (BCEC) was examined using reverse transcription-polymerase chain reaction analysis, which showed that there was no expression of rOat1 or rOat2, but moderate expression of rOat3. The expression of rOat3 in the BCEC was further confirmed by Western blotting. Immunohistochemical staining showed that rOat3 is located on the abluminal and, possibly, luminal membrane of the BCEC. The contribution of rOat3 to the efflux of para-aminohippuric acid (PAH) and benzylpenicillin (PCG), substrates of rOat3, from the cerebrum into the blood circulation across the BBB was evaluated using the Brain Efflux Index method. PAH and PCG were eliminated from the cerebrum with rate constants of 0.039 and 0.043 min Ϫ1 , respectively, and the elimination was saturated at high substrate concentrations. Taking account of the dilution in the brain, the K m values for the elimination of PAH and PCG were estimated to be 168 and 29 M, respectively. The efflux of PAH and PCG across the BBB was inhibited in a dose-dependent manner by unlabeled PCG and PAH, respectively. The K i value of PAH for the efflux of PCG was 106 M and that of PCG for the efflux of PAH was 58 M. These values were comparable with their K m values, suggesting that they share the same efflux mechanism at the BBB. Furthermore, cimetidine and pravastatin, which are also substrates and inhibitors of rOat3, significantly inhibited the efflux of PAH and PCG from the cerebrum. These results suggest that rOat3 is responsible for the elimination of PAH and PCG from the brain across the BBB.

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Expression and Functional Characterization of Rat Organic Anion Transporter 3 (rOat3) in the Choroid Plexus

Cited by 138 publications

References 30 publications

Organic anion transporters: discovery, pharmacology, regulation and roles in pathophysiology

Organic anion transporters: discovery, pharmacology, regulation and roles in pathophysiology

Use of Z310 cells as an in vitro blood–cerebrospinal fluid barrier model: Tight junction proteins and transport properties

Contribution of Organic Anion Transporter 3 (Slc22a8) to the Elimination of p-Aminohippuric Acid and Benzylpenicillin across the Blood-Brain Barrier

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