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Figure 7. Functional and pharmacological characterization of human nicotinate transporters, hSMCT1, SMCT2, and OAT10. The uptake of [14C]‐nicotinate (40 μmol/L) by SMCT1‐, SMCT2‐ or OAT10‐expressing oocytes was performed in ND96 medium (pH 7.4) at ~25°C. [14C]‐nicotinate transport properties of nicotinate transporters: voltage sensitivity and Na+‐dependence was examined by replacing extracellular NaCl by KCl or LiCl. Chloride ion‐dependence was also assessed; in 0 Cl− bath, NaCl was replaced by Na‐gluconate (Na‐Gluc), KCl by K‐gluconate (K‐Gluc), MgCl2 by Mg‐gluconate, and CaCl2 by Ca‐gluconate. (A) [14C]‐nicotinate transport properties of human SMCT1 (B) [14C]nicotinate transport properties of human SMCT2. *P < 0.001 compared with NaCl (ND96). (C) Inhibition of [14C]‐nicotinate uptake via SMCT1, and SMCT2 in the presence of uricosuric drugs added into the extracellular medium (pH 7.4) at the indicated concentrations. P < 0.01 compared with NaCl (ND96); NS, not significant. (D) [14C]‐nicotinate transport properties of human OAT10 in oocytes preloaded with PZA. *P < 0.001 compared with uptake in the absence of inhibitor; NS, not significant. (E) Inhibition of [14C]‐nicotinate uptake via OAT10 (preloaded with PZA) in the presence of uricosuric drugs added into the extracellular medium (pH 7.4) at the indicated concentrations. P < 0.01 compared with NaCl (ND96); NS, not significant. Tran, Benz, Prob, DMSO. Data are mean ± S.E. with n = 12–15. PZA, pyrazine carboxylate; Tran, Tranilast; Benz, Benzbromarone; Prob, probenecid; DMSO, dimethylsulfoxide. Image published in: Mandal AK et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |