Localization and functional characterization of the rat Oatp4c1 transporter in an in vitro cell system and rat tissues

Kuei-Ling Kuo; Haining Zhu; Patrick J McNamara; Markos Leggas

doi:10.1371/journal.pone.0039641

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Localization and functional characterization of the rat Oatp4c1 transporter in an in vitro cell system and rat tissues

Journal article

Open access

Peer reviewed

Localization and functional characterization of the rat Oatp4c1 transporter in an in vitro cell system and rat tissues

Kuei-Ling Kuo, Haining Zhu, Patrick J McNamara and Markos Leggas

PloS one, Vol.7(6), pp.e39641-e39641

2012

DOI: 10.1371/journal.pone.0039641

PMCID: PMC3387246

PMID: 22768102

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Abstract

The organic anion transporting polypeptide 4c1 (Oatp4c1) was previously identified as a novel uptake transporter predominantly expressed at the basolateral membrane in the rat kidney proximal tubules. Its functional role was suggested to be a vectorial transport partner of an apically-expressed efflux transporter for the efficient translocation of physiological substrates into urine, some of which were suggested to be uremic toxins. However, our in vitro studies with MDCKII cells showed that upon transfection rat Oatp4c1 polarizes to the apical membrane. In this report, we validated the trafficking and function of Oatp4c1 in polarized cell systems as well as its subcellular localization in rat kidney. Using several complementary biochemical, molecular and proteomic methods as well as antibodies amenable to immunohistochemistry, immunofluorescence, and immunobloting we investigated the expression pattern of Oatp4c1 in polarized cell systems and in the rat kidney. Collectively, these data demonstrate that rat Oatp4c1 traffics to the apical cell surface of polarized epithelium and localizes primarily in the proximal straight tubules, the S3 fraction of the nephron. Drug uptake studies in Oatp4c1-overexpressing cells demonstrated that Oatp4c1-mediated estrone-3-sulfate (E3S) uptake was pH-dependent and ATP-independent. These data definitively demonstrate the subcellular localization and histological location of Oatp4c1 and provide additional functional evidence that reconciles expression-function reports found in the literature.

Cell Line

Cell Polarity

Reproducibility of Results

Rats

Uremia - metabolism

Organ Specificity

Rats, Sprague-Dawley

Subcellular Fractions - metabolism

Organic Anion Transporters - metabolism

Protein Transport

Paraffin Embedding

Kidney - metabolism

Animals

Estrone - analogs & derivatives

Microvilli - metabolism

Estrone - metabolism

Dogs

Kinetics

Tissue Fixation

Hydrogen-Ion Concentration

Details

Title: Subtitle: Localization and functional characterization of the rat Oatp4c1 transporter in an in vitro cell system and rat tissues
Creators: Kuei-Ling Kuo - Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky
Haining Zhu
Patrick J McNamara
Markos Leggas
Resource Type: Journal article
Publication Details: PloS one, Vol.7(6), pp.e39641-e39641
DOI: 10.1371/journal.pone.0039641
PMID: 22768102
PMCID: PMC3387246
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science
Grant note: 8 P20 GM103486-09 / NIGMS NIH HHS P20 GM103486 / NIGMS NIH HHS 5P20RR020171-09 / NCRR NIH HHS P20 RR020171 / NCRR NIH HHS
Language: English
Date published: 2012
Academic Unit: Stead Family Department of Pediatrics; Neonatology; Internal Medicine
Record Identifier: 9984093312702771

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