Functional analysis of organic cation transporter 3 expressed in human placenta

Ryoko Sata, Hisakazu Ohtani, Masayuki Tsujimoto, Hideyasu Murakami, Noriko Koyabu, Takanori Nakamura, Takeshi Uchiumi, Michihiko Kuwano, Hideaki Nagata, Kiyomi Tsukimori, Hitoo Nakano, Yasufumi Sawada

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Abstract

The aim of this study is to investigate the placental transport mechanism of cationic compounds by comparison of the uptake of an organic cation into human placental basal membrane vesicles (BLMVs) with that into organic cation transporter 3 (OCT3)-expressing cells. Reverse transcription-polymerase chain reaction analysis demonstrated that OCT3 is the only OCT isoform expressed in the human placenta. The function of OCT3 was investigated by measuring the uptake of 1-methyl-4-phenylpyridinium (MPP+) into human embryonic kidney (HEK)293 cells stably expressing OCT3 (HEK/OCT3 cells). The OCT3-mediated uptake of MPP+ was sodium- and chloride-independent and saturable, with a Michaelis constant (Km) of 82.5 μM. The OCT3-mediated uptake was inhibited by various cationic drugs in a concentration-dependent manner but not by anionic compounds, such as p-aminohippuric acid and captopril, or a zwitterion, carnitine. Western blotting analysis of membrane vesicles prepared from human term placenta revealed that OCT3 is expressed only in BLMVs but not in microvillous membrane vesicles. The uptake of MPP+ into BLMVs was membrane potential-dependent and saturable, with a Km value of 51.8 μM, which is similar to that in HEK293/OCT3 cells. The inhibitory spectrum of various compounds on MPP+ uptake by BLMVs was also similar to that in HEK293/OCT3 cells. These results suggest that OCT3 is expressed on the basal membrane of human trophoblast cells and plays an important role in the placental transport of cationic compounds.

Original languageEnglish
Pages (from-to)888-895
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume315
Issue number2
DOIs
Publication statusPublished - Nov 1 2005

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Placenta
Cations
Membranes
1-Methyl-4-phenylpyridinium
p-Aminohippuric Acid
Kidney
Carnitine
Captopril
Trophoblasts
Sodium Chloride
Membrane Potentials
Reverse Transcription
Protein Isoforms
Western Blotting
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

Sata, R., Ohtani, H., Tsujimoto, M., Murakami, H., Koyabu, N., Nakamura, T., ... Sawada, Y. (2005). Functional analysis of organic cation transporter 3 expressed in human placenta. Journal of Pharmacology and Experimental Therapeutics, 315(2), 888-895. https://doi.org/10.1124/jpet.105.086827

Functional analysis of organic cation transporter 3 expressed in human placenta. / Sata, Ryoko; Ohtani, Hisakazu; Tsujimoto, Masayuki; Murakami, Hideyasu; Koyabu, Noriko; Nakamura, Takanori; Uchiumi, Takeshi; Kuwano, Michihiko; Nagata, Hideaki; Tsukimori, Kiyomi; Nakano, Hitoo; Sawada, Yasufumi.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 315, No. 2, 01.11.2005, p. 888-895.

Research output: Contribution to journalArticle

Sata, R, Ohtani, H, Tsujimoto, M, Murakami, H, Koyabu, N, Nakamura, T, Uchiumi, T, Kuwano, M, Nagata, H, Tsukimori, K, Nakano, H & Sawada, Y 2005, 'Functional analysis of organic cation transporter 3 expressed in human placenta', Journal of Pharmacology and Experimental Therapeutics, vol. 315, no. 2, pp. 888-895. https://doi.org/10.1124/jpet.105.086827
Sata, Ryoko ; Ohtani, Hisakazu ; Tsujimoto, Masayuki ; Murakami, Hideyasu ; Koyabu, Noriko ; Nakamura, Takanori ; Uchiumi, Takeshi ; Kuwano, Michihiko ; Nagata, Hideaki ; Tsukimori, Kiyomi ; Nakano, Hitoo ; Sawada, Yasufumi. / Functional analysis of organic cation transporter 3 expressed in human placenta. In: Journal of Pharmacology and Experimental Therapeutics. 2005 ; Vol. 315, No. 2. pp. 888-895.
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