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ATP-Binding Cassette Transporter G2 Mediates the Efflux of Phototoxins on the Luminal Membrane of Retinal Capillary Endothelial Cells

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Purpose

The purpose of this study was to clarify the localization and function of the ATP-binding cassette transporter G2 (ABCG2; BCRP/MXR/ABCP) in retinal capillary endothelial cells, which form the inner blood–retinal barrier, as an efflux transport system.

Methods

The expression was determined by reverse transcriptase polymerase chain reaction and Western blotting. The localization was identified by immunostaining. The transport function of ABCG2 was measured by flow cytometry.

Results

Western blotting indicated that ABCG2 was expressed as a glycosylated disulfide-linked complex in the mouse retina and in peripheral tissues, including liver, kidney, and small intestine. Double immunolabeling of ABCG2 and glucose transporter 1 suggested that ABCG2 was localized on the luminal membrane of mouse retinal capillary endothelial cells. ABCG2 mRNA and protein were found to be expressed in a conditionally immortalized rat retinal capillary endothelial cell line, TR-iBRB, and rat retina. Treatment with Ko143, an ABCG2 inhibitor, restored the accumulation of pheophorbide a and protoporphyrin IX in TR-iBRB cells.

Conclusion

ABCG2 is expressed on the luminal membrane of retinal capillary endothelial cells, where ABCG2 acts as the efflux transporter for photosensitive toxins such as pheophorbide a and protoporphyrin IX. ABCG2 could play an important role at the inner blood–retinal barrier in restricting the distribution of phototoxins and xenobiotics in retinal tissue.

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Abbreviations

ABC:

ATP-binding cassette

GLUT1:

glucose transporter 1

P-gp:

P-glycoprotein

PhA:

pheophorbide a

PPIX:

protoporphyrin IX

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Acknowledgments

We would like to thank Mr. N. Kimura for technical assistance and Ms. N. Funayama for secretarial assistance. We would like to thank Drs. Ken-ichi Hosoya and Masatoshi Tomi of University of Toyama for valuable suggestions. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) and Japan Society for the Promotion of Science (JSPS) and a 21st Century Center of Excellence (COE) Program from JSPS. It was also supported in part by the Industrial Technology Research Grant Program from the New Energy and Industrial Technology Development Organization (NEDO) of Japan.

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Authors and Affiliations

  1. Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan

    Tomoko Asashima, Satoko Hori, Sumio Ohtsuki, Masanori Tachikawa & Tetsuya Terasaki

  2. New Industry Creation Hatchery Center, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8579, Japan

    Tomoko Asashima, Satoko Hori, Sumio Ohtsuki & Tetsuya Terasaki

  3. CREST and SORST of Japan Science and Technology Agency (JST),  , Japan

    Tomoko Asashima, Satoko Hori, Sumio Ohtsuki & Tetsuya Terasaki

  4. Department of Anatomy, Hokkaido University School of Medicine, Sapporo, 060-8638, Japan

    Masahiko Watanabe

  5. Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan

    Chisato Mukai, Shinji Kitagaki & Naoki Miyakoshi

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  1. Tomoko Asashima
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Correspondence to Tetsuya Terasaki.

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Asashima, T., Hori, S., Ohtsuki, S. et al. ATP-Binding Cassette Transporter G2 Mediates the Efflux of Phototoxins on the Luminal Membrane of Retinal Capillary Endothelial Cells. Pharm Res 23, 1235–1242 (2006). https://doi.org/10.1007/s11095-006-0067-2

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