| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Laboratory of Pharmacology and Chemistry, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709
The organic anion transporter, rROAT1, is a dicarboxylate/organic anion exchanger, a function associated with the basolateral membrane in rat proximal tubule. To directly establish the subcellular localization of rROAT1 in renal epithelia, we made a rROAT1-green fluorescent protein (GFP) fusion construct (rROAT1-GFP). Plasma membrane-associated fluorescence was observed in rROAT1-GFP-expressing Xenopus oocytes examined by confocal microscopy. Uptake of 3H-labeled p-aminohippurate (PAH) increased 2.5-fold in rROAT1-GFP-expressing Xenopus oocytes, and this increase was abolished by 1 mM probenecid. Thus the construct was capable of specific organic anion transport. Cultured renal epithelial cell lines (MDCK and LLC-PK1) transfected with the vector pEGFP-C3 showed a diffuse, evenly distributed cytoplasmic signal. However, when transfected with pEGFP-C3/rROAT1 (vector coding for rROAT1-GFP), both cell lines showed predominantly plasma membrane fluorescence. The expression and distribution of rROAT1-GFP in intact renal proximal tubules was also investigated. Isolated killifish (Fundulus heteroclitus) renal tubules transfected with pEGFP-C3/rROAT1 showed marked basal and lateral membrane-associated fluorescence, but no detectable signal in the nucleus or the apical pole of tubule cells. Tubules transfected with pEGFP-C3 showed diffuse cytoplasmic fluorescence. Function of the rROAT1-GFP construct was demonstrated in transfected killifish tubules by fluorescein transport assay. These results demonstrate the basolateral subcellular localization of rROAT1 in polarized renal epithelia and validate a new technique for localizing cloned transporters within intact renal tubules.
kidney; killifish; Xenopus; transfection; Madin-Darby canine kidney cells; LLC-PK1 cells; green flourescent protein
This article has been cited by other articles:
|
|
 |
|
  G. W. Schnabolk, G. L. Youngblood, and D. H. Sweet Transport of estrone sulfate by the novel organic anion transporter Oat6 (Slc22a20) Am J Physiol Renal Physiol, August 1, 2006; 291(2): F314 - F321. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. L. Youngblood and D. H. Sweet Identification and functional assessment of the novel murine organic anion transporter Oat5 (Slc22a19) expressed in kidney Am J Physiol Renal Physiol, August 1, 2004; 287(2): F236 - F244. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. H. Wright and W. H. Dantzler Molecular and Cellular Physiology of Renal Organic Cation and Anion Transport Physiol Rev, July 1, 2004; 84(3): 987 - 1049. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. W. Beyenbach Kidneys sans glomeruli Am J Physiol Renal Physiol, May 1, 2004; 286(5): F811 - F827. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Aslamkhan, Y.-H. Han, R. Walden, D. H. Sweet, and J. B. Pritchard Stoichiometry of organic anion/dicarboxylate exchange in membrane vesicles from rat renal cortex and hOAT1-expressing cells Am J Physiol Renal Physiol, October 1, 2003; 285(4): F775 - F783. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Mangravite, G. Xiao, and K. M. Giacomini Localization of human equilibrative nucleoside transporters, hENT1 and hENT2, in renal epithelial cells Am J Physiol Renal Physiol, May 1, 2003; 284(5): F902 - F910. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Sweet, L. M. S. Chan, R. Walden, X.-P. Yang, D. S. Miller, and J. B. Pritchard Organic anion transporter 3 (Slc22a8) is a dicarboxylate exchanger indirectly coupled to the Na+ gradient Am J Physiol Renal Physiol, April 1, 2003; 284(4): F763 - F769. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. G. Aslamkhan, Y.-H. Han, X.-P. Yang, R. K. Zalups, and J. B. Pritchard Human Renal Organic Anion Transporter 1-Dependent Uptake and Toxicity of Mercuric-Thiol Conjugates in Madin-Darby Canine Kidney Cells Mol. Pharmacol., March 1, 2003; 63(3): 590 - 596. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. H. Sweet, D. S. Miller, J. B. Pritchard, Y. Fujiwara, D. R. Beier, and S. K. Nigam Impaired Organic Anion Transport in Kidney and Choroid Plexus of Organic Anion Transporter 3 (Oat3 (Slc22a8)) Knockout Mice J. Biol. Chem., July 19, 2002; 277(30): 26934 - 26943. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Nagata, H. Kusuhara, H. Endou, and Y. Sugiyama Expression and Functional Characterization of Rat Organic Anion Transporter 3 (rOat3) in the Choroid Plexus Mol. Pharmacol., May 1, 2002; 61(5): 982 - 988. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Lee, S. Dallas, M. Hong, and R. Bendayan Drug Transporters in the Central Nervous System: Brain Barriers and Brain Parenchyma Considerations Pharmacol. Rev., December 1, 2001; 53(4): 569 - 596. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Sweet, D. S. Miller, and J. B. Pritchard Ventricular Choline Transport. A ROLE FOR ORGANIC CATION TRANSPORTER 2 EXPRESSED IN CHOROID PLEXUS J. Biol. Chem., November 2, 2001; 276(45): 41611 - 41619. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. S. Miller Nucleoside Phosphonate Interactions with Multiple Organic Anion Transporters in Renal Proximal Tubule J. Pharmacol. Exp. Ther., November 1, 2001; 299(2): 567 - 574. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Pombrio, A. Giangreco, L. Li, M. F. Wempe, M. W. Anders, D. H. Sweet, J. B. Pritchard, and N. Ballatori Mercapturic Acids (N-Acetylcysteine S-Conjugates) as Endogenous Substrates for the Renal Organic Anion Transporter-1 Mol. Pharmacol., November 1, 2001; 60(5): 1091 - 1099. [Abstract] [Full Text]
|
|
|
|
|
|
D. H. Sweet, K. T. Bush, and S. K. Nigam The organic anion transporter family: from physiology to ontogeny and the clinic Am J Physiol Renal Physiol, August 1, 2001; 281(2): F197 - F205. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. M. Mangravite, J. H. Lipschutz, K. E. Mostov, and K. M. Giacomini Localization of GFP-tagged concentrative nucleoside transporters in a renal polarized epithelial cell line Am J Physiol Renal Physiol, May 1, 2001; 280(5): F879 - F885. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Sweet, D. S. Miller, and J. B. Pritchard Basolateral localization of organic cation transporter 2 in intact renal proximal tubules Am J Physiol Renal Physiol, November 1, 2000; 279(5): F826 - F834. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Shuprisha, S. H. Wright, and W. H. Dantzler Method for measuring luminal efflux of fluorescent organic compounds in isolated, perfused renal tubules Am J Physiol Renal Physiol, November 1, 2000; 279(5): F960 - F964. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. You, K. Kuze, R. A. Kohanski, K. Amsler, and S. Henderson Regulation of mOAT-mediated Organic Anion Transport by Okadaic Acid and Protein Kinase C in LLC-PK1 Cells J. Biol. Chem., March 31, 2000; 275(14): 10278 - 10284. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. B. Pritchard, D. H. Sweet, D. S. Miller, and R. Walden Mechanism of Organic Anion Transport across the Apical Membrane of Choroid Plexus J. Biol. Chem., November 19, 1999; 274(47): 33382 - 33387. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Visit Other APS Journals Online |
