Roles of inner blood-retinal barrier organic anion transporter 3 in the vitreous/retina-to-blood efflux transport of p-aminohippuric acid, benzylpenicillin, and 6-mercaptopurine

Ken Ichi Hosoya, Akihide Makihara, Yuki Tsujikawa, Daisuke Yoneyama, Shinobu Mori, Tetsuya Terasaki, Shin Ichi Akanuma, Masatoshi Tomi, Masanori Tachikawa

Research output: Contribution to journalArticle

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Abstract

The purpose of the present study was to characterize rat organic anion transporter (Oat) 3 (Oat3, Slc22a8) in the efflux transport at the inner blood-retinal barrier (BRB). Reverse transcription-polymerase chain reaction analysis showed that rat (r) Oat3 mRNA is expressed in retinal vascular endothelial cells (RVECs), but not rOat1 and rOat2 mRNA. The expression of Oat3 in the retina and human cultured retinal endothelial cells was further confirmed by Western blot analysis. Immunohisto-chemical staining in RVECs showed that rOat3 is colocalized with glucose transporter 1, but not P-glycoprotein, suggesting that rOat3 is possibly located at the abluminal membrane of the RVEC. The contribution of rOat3 to the efflux of [3H]p-aminohippuric acid ([3H]PAH), [3H]benzylpenicillin ([3H]PCG), and [14C]6-mercaptopurine ([14C]6-MP), substrates of rOat3, from the vitreous humor/retina to the circulating blood across the inner BRB was evaluated using the microdialysis method. [3H]PAH, [ 3H]PCG, [14C]6-MP, and [14C] or [ 3H]D-mannitol, a bulk flow marker, were biexponentially eliminated from the vitreous humor after vitreous bolus injection. The elimination rate constant of [3H]PAH, [3H]PCG, and [14C]6-MP during the terminal phase was approximately 2-fold greater than that of D-mannitol. This efflux transport was reduced in the retinal presence of probenecid, PAH, and PCG, whereas it was not inhibited by digoxin. In conclusion, rOat3 is expressed at the inner BRB and involved in the vitreous humor/retina-to-blood transport of PAH, PCG, and 6-MP. This transport system is one mechanism to limit the retinal distribution of PAH, PCG, and 6-MP.

Original languageEnglish
Pages (from-to)87-93
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume329
Issue number1
DOIs
Publication statusPublished - 2009 Apr
Externally publishedYes

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Blood-Retinal Barrier
Organic Anion Transporters
p-Aminohippuric Acid
6-Mercaptopurine
Penicillin G
Vitreous Body
Retinal Vessels
Retina
Endothelial Cells
Mannitol
Far-Western Blotting
Probenecid
Messenger RNA
Facilitative Glucose Transport Proteins
Digoxin
Microdialysis
Reverse Transcription
Staining and Labeling
Polymerase Chain Reaction
Injections

ASJC Scopus subject areas

  • Pharmacology
  • Molecular Medicine

Cite this

Roles of inner blood-retinal barrier organic anion transporter 3 in the vitreous/retina-to-blood efflux transport of p-aminohippuric acid, benzylpenicillin, and 6-mercaptopurine. / Hosoya, Ken Ichi; Makihara, Akihide; Tsujikawa, Yuki; Yoneyama, Daisuke; Mori, Shinobu; Terasaki, Tetsuya; Akanuma, Shin Ichi; Tomi, Masatoshi; Tachikawa, Masanori.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 329, No. 1, 04.2009, p. 87-93.

Research output: Contribution to journalArticle

Hosoya, Ken Ichi ; Makihara, Akihide ; Tsujikawa, Yuki ; Yoneyama, Daisuke ; Mori, Shinobu ; Terasaki, Tetsuya ; Akanuma, Shin Ichi ; Tomi, Masatoshi ; Tachikawa, Masanori. / Roles of inner blood-retinal barrier organic anion transporter 3 in the vitreous/retina-to-blood efflux transport of p-aminohippuric acid, benzylpenicillin, and 6-mercaptopurine. In: Journal of Pharmacology and Experimental Therapeutics. 2009 ; Vol. 329, No. 1. pp. 87-93.
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abstract = "The purpose of the present study was to characterize rat organic anion transporter (Oat) 3 (Oat3, Slc22a8) in the efflux transport at the inner blood-retinal barrier (BRB). Reverse transcription-polymerase chain reaction analysis showed that rat (r) Oat3 mRNA is expressed in retinal vascular endothelial cells (RVECs), but not rOat1 and rOat2 mRNA. The expression of Oat3 in the retina and human cultured retinal endothelial cells was further confirmed by Western blot analysis. Immunohisto-chemical staining in RVECs showed that rOat3 is colocalized with glucose transporter 1, but not P-glycoprotein, suggesting that rOat3 is possibly located at the abluminal membrane of the RVEC. The contribution of rOat3 to the efflux of [3H]p-aminohippuric acid ([3H]PAH), [3H]benzylpenicillin ([3H]PCG), and [14C]6-mercaptopurine ([14C]6-MP), substrates of rOat3, from the vitreous humor/retina to the circulating blood across the inner BRB was evaluated using the microdialysis method. [3H]PAH, [ 3H]PCG, [14C]6-MP, and [14C] or [ 3H]D-mannitol, a bulk flow marker, were biexponentially eliminated from the vitreous humor after vitreous bolus injection. The elimination rate constant of [3H]PAH, [3H]PCG, and [14C]6-MP during the terminal phase was approximately 2-fold greater than that of D-mannitol. This efflux transport was reduced in the retinal presence of probenecid, PAH, and PCG, whereas it was not inhibited by digoxin. In conclusion, rOat3 is expressed at the inner BRB and involved in the vitreous humor/retina-to-blood transport of PAH, PCG, and 6-MP. This transport system is one mechanism to limit the retinal distribution of PAH, PCG, and 6-MP.",
author = "Hosoya, {Ken Ichi} and Akihide Makihara and Yuki Tsujikawa and Daisuke Yoneyama and Shinobu Mori and Tetsuya Terasaki and Akanuma, {Shin Ichi} and Masatoshi Tomi and Masanori Tachikawa",
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AU - Makihara, Akihide

AU - Tsujikawa, Yuki

AU - Yoneyama, Daisuke

AU - Mori, Shinobu

AU - Terasaki, Tetsuya

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