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Am J Physiol Cell Physiol 288: C290-C303, 2005. First published October 13, 2004; doi:10.1152/ajpcell.00053.2004
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Multiple pathways for cationic amino acid transport in rat thyroid epithelial cell line PC Cl3

Tiziano Verri,1 Cinzia Dimitri,1 Sonia Treglia,2 Fabio Storelli,2 Stefania De Micheli,2 Luca Ulianich,3,4 Pasquale Vito,3,5,6 Santo Marsigliante,1 Carlo Storelli,1 and Bruno Di Jeso2

1Laboratory of General Physiology and 2Laboratory of General Pathology, Department of Biological and Environmental Sciences and Technologies, University of Lecce, Lecce; 3Department of Cellular and Molecular Biology and Pathology "L. Califano" and 5BioGeM Consortium, Federico II University of Naples, Naples; 4Institute of Endocrinology and Experimental Oncology "G. Salvatore," Italian National Research Council, Naples; and 6Laboratory of Molecular Genetics, Department of Biological and Environmental Sciences, University of Sannio, Benevento, Italy

Submitted 27 January 2004 ; accepted in final form 10 October 2004

Information regarding cationic amino acid transport systems in thyroid is limited to Northern blot detection of y+LAT1 mRNA in the mouse. This study investigated cationic amino acid transport in PC cell line clone 3 (PC Cl3 cells), a thyroid follicular cell line derived from a normal Fisher rat retaining many features of normal differentiated follicular thyroid cells. We provide evidence that in PC Cl3 cells plasmalemmal transport of cationic amino acids is Na+ independent and occurs, besides diffusion, with the contribution of high-affinity, carrier-mediated processes. Carrier-mediated transport is via y+, y+L, and b0,+ systems, as assessed by L-arginine uptake and kinetics, inhibition of L-arginine transport by N-ethylmaleimide and neutral amino acids, and L-cystine transport studies. y+L and y+ systems account for the highest transport rate (with y+L > y+) and b0,+ for a residual fraction of the transport. Uptake data correlate to expression of the genes encoding for CAT-1, CAT-2B, 4F2hc, y+LAT1, y+LAT2, rBAT, and b0,+AT, an expression profile that is also shown by the rat thyroid gland. In PC Cl3 cells cationic amino acid uptake is under TSH and/or cAMP control (with transport increasing with increasing TSH concentration), and upregulation of CAT-1, CAT-2B, 4F2hc/y+LAT1, and rBAT/b0,+AT occurs at the mRNA level under TSH stimulation. Our results provide the first description of an expression pattern of cationic amino acid transport systems in thyroid cells. Furthermore, we provide evidence that extracellular L-arginine is a crucial requirement for normal PC Cl3 cell growth and that long-term L-arginine deprivation negatively influences CAT-2B expression, as it correlates to reduction of CAT-2B mRNA levels.

cationic amino acid transporters; heteromeric amino acid transporters; system y+; system y+L; system b0,+; thyrotropin; L-arginine



Address for reprint requests and other correspondence: T. Verri, Laboratory of General Physiology, Dept. of Biological and Environmental Sciences and Technologies, Univ. of Lecce, Strada Provinciale Lecce-Monteroni, I-73100 Lecce, Italy (E-mail: physiol{at}ultra5.unile.it)




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