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In Vitro Polarized Transport of L-Phenylalanine in Human Nasal Epithelium and Partial Characterization of the Amino Acid Transporters Involved

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Abstract

Purpose. The purpose of this study was to provide functional and molecular evidence to support the existence of large neutral amino acid transporters in human nasal epithelium using nasal primary cell culture model.

Methods. L-Phenylalanine was used as a model substrate to characterize carrier-mediated permeation of amino acids across human nasal epithelium. The influence of temperature, concentration, other amino acids, metabolic/transport inhibitors, and polarity/stereo-selectivity on transport of the model compound was investigated. Reverse transcriptase polymerase chain reaction was used for molecular characterization of the existence of the transporters.

Results. The transport of L-phenylalanine across the human nasal epithelium was polarized (apical → basolateral >> basolateral → apical), saturable (K m = 1.23 mM; V max = 805.1 nmol/mg protein/min) and stereo-selective (permeation of L-phenylalanine >> D-Phenyl- alanine). Its permeation was significantly (<0.05) reduced by cationic, small and large neutral amino acids, oubain, amiloride, sodium-free medium, and temperature lowering. Reverse transcriptase polymerase chain reaction revealed the presence of the broad-scope cationic-dependent amino acid transporter gene (y+LAT-2) in the human nasal epithelium.

Conclusions. Based on the results of this study, one may postulate that the human nasal epithelium expresses L-amino acid transporters. More studies are necessary for detailed characterization of the transporters.

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Authors and Affiliations

  1. Laboratory for Pharmacotechnology and Biopharmacy, K.U. Leuven, Campus Gasthuisberg O&N, Herestraat 49, B-3000, Leuven, Belgium

    Remigius Agu, Hoang Vu Dang, Renaat Kinget & Norbert Verbeke

  2. Department of Pharmaceutical Sciences, College of Pharmacy 518, University of Kentucky, Rose Street Lexington, Kentucky, 40536-0082

    Remigius Agu

  3. Department of Pharmaceutical Sciences, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, 27 Taylor Street, Glasgow, G40NR

    Hoang Vu Dang

  4. Laboratory for Experimental Otorhinolaryngology, K.U. Leuven, Campus Gasthuisberg O&N, Herestraat 49, B-3000, Leuven, Belgium

    Mark Jorissen & Tom Willems

  5. Laboratory for Medical Biochemistry, K.U. Leuven, Campus Gasthuisberg O&N, Herestraat 49, B-3000, Leuven, Belgium

    Sandy Vandoninck, Johan Van Lint & Jackie V. Vandenheede

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  1. Remigius Agu
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  2. Hoang Vu Dang
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  8. Renaat Kinget
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  9. Norbert Verbeke
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Correspondence to Norbert Verbeke.

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Agu, R., Dang, H.V., Jorissen, M. et al. In Vitro Polarized Transport of L-Phenylalanine in Human Nasal Epithelium and Partial Characterization of the Amino Acid Transporters Involved. Pharm Res 20, 1125–1132 (2003). https://doi.org/10.1023/A:1025028410131

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