title: In vitro polarized transport of L-phenylalanine in human nasal epithelium and partial characterization of the amino acid transporters involved.
abstract: OBJECTIVE
  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 (Km = 1.23 mM; Vmax = 805.1 nmol/mg protein/min) and stereo-selective (permeation of L-phenylalanine >> D-Phenylalanine). 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.
authors.affiliation: Laboratory for Pharmacotechnology and Biopharmacy, K.U. Leuven, Campus Gasthuisberg O&N, Herestraat 49, B-3000 Leuven, Belgium.
authors.forename: Remigius
authors.initials: R
authors.lastname: Agu
authors.forename: Hoang Vu
authors.initials: HV
authors.lastname: Dang
authors.forename: Mark
authors.initials: M
authors.lastname: Jorissen
authors.forename: Tom
authors.initials: T
authors.lastname: Willems
authors.forename: Sandy
authors.initials: S
authors.lastname: Vandoninck
authors.forename: Johan
authors.initials: J
authors.lastname: Van Lint
authors.forename: Jackie V
authors.initials: JV
authors.lastname: Vandenheede
authors.forename: Renaat
authors.initials: R
authors.lastname: Kinget
authors.forename: Norbert
authors.initials: N
authors.lastname: Verbeke
mesh_terms: D026905:Amino Acid Transport Systems / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active; D002478:Cells, Cultured; D006801:Humans; D009297:Nasal Mucosa / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature
chemical_list: D026905:Amino Acid Transport Systems; D012333:RNA, Messenger; D010649:Phenylalanine