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1 Laboratorio di Genetica Molecolare, 2 Clinica Pediatrica, 3 Divisione di Pneumologia, and 4 Laboratorio di Biologia Molecolare, Istituto Giannina Gaslini, 16148 Genoa, Italy
We performed Ussing chamber experiments on cultured human bronchial epithelial cells to look for the presence of electrogenic dibasic amino acid transport. Apical but not basolateral L-arginine (10-1,000 µM) increased the short-circuit current. Maximal effect and EC50 were ~3.5 µA/cm2 and 80 µM, respectively, in cells from normal subjects and cystic fibrosis patients. The involvement of nitric oxide was ruled out because a nitric oxide synthase inhibitor (NG-nitro-L-arginine methyl ester) did not decrease the arginine-dependent current. Apical L-lysine, L-alanine, and L-proline, but not aspartic acid, were also effective in increasing the short-circuit current, with EC50 values ranging from 26 to 971 µM. Experiments performed with radiolabeled arginine demonstrated the presence of an Na+-dependent concentrative transporter on the apical membrane of bronchial cells. This transporter could be important in vivo to maintain a low amino acid concentration in the fluid covering the airway surface.
airway epithelium; arginine transport; nitric oxide; pulmonary surfactant; cystic fibrosis
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