Abstract
Cadmium (Cd)-intoxicated experimental animals exhibit impaired renal secretion of organic anions (OA) and cations (OC), indicating their transporters (Oats and Octs) in the proximal tubule (PT) basolateral membrane as possible targets of Cd. To correlate transport data from the literature with the expression of relevant transporters, we performed immunochemical and RT-PCR studies of renal Oats and Octs in the subchronic (treatment with CdCl2; 2 mg Cd/kg b.m./day, for 2 weeks) and acute (treatment with Cd-metallothionein (CdMT); 0.4 mg Cd/kg b.m., 6 or 12 h before killing) models of Cd nephrotoxicity. In the subchronic model, PT exhibited a minor loss of basolateral invaginations and overall unchanged expression of Na+/K+-ATPase and GAPDH proteins and mRNAs, while the expression of Oat and Oct proteins and their mRNAs was strongly downregulated. In the acute model, a time-related redistribution of basolateral transporters to the intracellular vesicular compartment was a major finding. However, 6 h following CdMT treatment, the total abundance of Oat and Oct proteins in the renal tissue remained unchanged, the expression of mRNAs decreased only for Oats, while a limited Oat1 and Na+/K+-ATPase immunoreactivity in the PT apical membrane indicated loss of cell polarity. As tested in rats treated with colchicine, the observed loss/redistribution of basolateral transporters in both models may be independent on microtubules. Therefore, the diminished renal secretion of OA and OC via PT in Cd nephrotoxicity may result from (a) limited loss of secretory surface (basolateral invaginations), (b) selective loss of Oats and Octs, and (c) loss of cell polarity.
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Abbreviations
- BBM:
-
Brush border membrane
- BLM:
-
Basolateral membrane
- Cd:
-
Cadmium
- Cd-NTX:
-
Cadmium-induced nephrotoxicity
- CdMT:
-
Cadmium-metallothionein complex
- Na+/K+-ATPase:
-
Sodium-potassium ATPase
- OA:
-
Organic anion
- OAT/Oat:
-
Organic anion transporter
- OC:
-
Organic cation
- OCT/Oct:
-
Organic cation transporter
- PAH:
-
p-Aminohippurate
- PT:
-
Proximal tubule
- Sglt1 and Sglt2:
-
Sodium-d-glucose cotransporters 1 and 2
- TALH:
-
Thick ascending limb of Henle
- TCM:
-
Total cell membranes
- TEA:
-
Tetraethylammonium
- WB:
-
Western blotting
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Acknowledgments
The authors thank Mrs. Eva Heršak for technical assistance, and Prof. Dr. Hermann Koepsell, Institute of Anatomy and Cell Biology, University of Würzburg, Germany, for generous donation of the Oct1, Oct2 and Sglt1 and Sglt2 antibodies. This work was supported by Grant 022-0222148-2146 from Ministry for Science, Education and Sports, Republic of Croatia (I.S.).
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The authors declare no conflicts of interest in this study.
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The manuscript does not contain clinical studies or patient data. All experiments were performed on experimental animals (rats) in conformity with the highest standards of international recommendations and rules on animal rights and were approved by the Institutional Ethic Committee.
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Ljubojević, M., Breljak, D., Herak-Kramberger, C.M. et al. Expression of basolateral organic anion and cation transporters in experimental cadmium nephrotoxicity in rat kidney. Arch Toxicol 90, 525–541 (2016). https://doi.org/10.1007/s00204-015-1450-8
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DOI: https://doi.org/10.1007/s00204-015-1450-8