Abstract
Uncoupling protein (UCP)-2 and -3 are ubiquitously expressed throughout the body but there is currently no information regarding the expression and distribution of the different UCP isoforms in the kidney. Due to the known cross-reactivity of the antibodies presently available for detection of UCP-2 and -3 proteins, we measured the mRNA expression of UCP-1, -2 and -3 in the rat kidney in order to detect the kidney-specific UCP isoforms. Thereafter, we determined the intrarenal distribution of the detected UCP isoforms using immunohistochemistry. Thereafter, we compared the protein levels in control and streptozotocin-induced diabetic rats using Western blot. Expressions of the UCP isoforms were also performed in brown adipose tissue and heart as positive controls for UCP-1 and 3, respectively.
UCP-2 mRNA was the only isoform detected in the kidney. UCP-2 protein expression in the kidney cortex was localized to proximal tubular cells, but not glomerulus or distal nephron. In the medulla, UCP-2 was localized to cells of the medullary thick ascending loop of Henle, but not to the vasculature or parts of the nephron located in the inner medulla. Western blot showed that diabetic kidneys have about 2.5-fold higher UCP-2 levels compared to controls.
In conclusion, UCP-2 is the only isoform detectable in the kidney and UCP-2 protein can be detected in proximal tubular cells and cells of the medullary thick ascending loop of Henle. Furthermore, diabetic rats have increased UCP-2 levels compared to controls, but the mechanisms underlying this increase and its consequences warrants further studies.
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References
T. Nishikawa, D. Edelstein, and M. Brownlee, The missing link: a single unifying mechanism for diabetic complications, Kidney Int Suppl 77(S26-30 (2000).
R. P. Robertson, Chronic oxidative stress as a central mechanism for glucose toxicity in pancreatic islet beta cells in diabetes, J Biol Chem 279(41), 42351-4 (2004).
M. Brownlee, Biochemistry and molecular cell biology of diabetic complications, Nature 414(6865), 813-20 (2001).
B. B. Lowell, and G. I. Shulman, Mitochondrial dysfunction and type 2 diabetes, Science 307(5708), 384-7 (2005).
S. S. Korshunov, V. P. Skulachev, and A. A. Starkov, High protonic potential actuates a mechanism of production of reactive oxygen species in mitochondria, FEBS Lett 416(1), 15-8 (1997).
S. S. Liu, Generating, partitioning, targeting and functioning of superoxide in mitochondria, Biosci Rep 17(3), 259-72 (1997).
M. D. Brand, Uncoupling to survive? The role of mitochondrial inefficiency in ageing, Exp Gerontol 35(6-7), 811-20 (2000).
T. Nishikawa, D. Edelstein, X. L. Du, S. Yamagishi, T. Matsumura, Y. Kaneda, M. A. Yorek, D. Beebe, P. J. Oates, H. P. Hammes, I. Giardino, and M. Brownlee, Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage, Nature 404(6779), 787-90 (2000).
P. Jezek, Possible physiological roles of mitochondrial uncoupling proteins–UCPn, Int J Biochem Cell Biol 34(10), 1190-206 (2002).
A. G. Dulloo, and S. Samec, Uncoupling proteins: their roles in adaptive thermogenesis and substrate metabolism reconsidered, Br J Nutr 86(2), 123-39 (2001).
A. Negre-Salvayre, C. Hirtz, G. Carrera, R. Cazenave, M. Troly, R. Salvayre, L. Penicaud, and L. Casteilla, A role for uncoupling protein-2 as a regulator of mitochondrial hydrogen peroxide generation, Faseb J 11(10), 809-15 (1997).
C. Duval, A. Negre-Salvayre, A. Dogilo, R. Salvayre, L. Penicaud, and L. Casteilla, Increased reactive oxygen species production with antisense oligonucleotides directed against uncoupling protein 2 in murine endothelial cells, Biochem Cell Biol 80(6), 757-64 (2002).
K. S. Echtay, E. Winkler, K. Frischmuth, and M. Klingenberg, Uncoupling proteins 2 and 3 are highly active H(+) transporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone), Proc Natl Acad Sci U S A 98(4), 1416-21 (2001).
A. M. Vincent, J. A. Olzmann, M. Brownlee, W. I. Sivitz, and J. W. Russell, Uncoupling proteins prevent glucose-induced neuronal oxidative stress and programmed cell death, Diabetes 53(3), 726-34 (2004).
F. Palm, J. Cederberg, P. Hansell, P. Liss, and P. O. Carlsson, Reactive oxygen species cause diabetesinduced decrease in renal oxygen tension, Diabetologia 46(8), 1153-60 (2003).
M. Friederich, J. Olerud, A. Fasching, P. Liss, P. Hansell, and F. Palm, Uncoupling protein 2 in diabetic kidneys: Increased protein expression correlates to increased non-transport related oxygen consumption, Adv Exp Med Biol In press((2007).
A. Tojo, M. Kimoto, and C. S. Wilcox, Renal expression of constitutive NOS and DDAH: separate effects of salt intake and angiotensin, Kidney Int 58(5), 2075-83 (2000).
F. Palm, and P. O. Carlsson, Thick ascending tubular cells in the loop of Henle: regulation of electrolyte homeostasis, Int J Biochem Cell Biol 37(8), 1554-9 (2005).
F. Palm, P. Hansell, G. Ronquist, A. Waldenstrom, P. Liss, and P. O. Carlsson, Polyol-pathway-dependent disturbances in renal medullary metabolism in experimental insulin-deficient diabetes mellitus in rats, Diabetologia 47(7), 1223-31 (2004).
A. Korner, A. C. Eklof, G. Celsi, and A. Aperia, Increased renal metabolism in diabetes. Mechanism and functional implications, Diabetes 43(5), 629-33 (1994).
F. Palm, L. Nordquist, and D. G. Buerk, Nitric oxide in the kidney; direct measurements of bioavailable renal nitric oxide, Adv Exp Med Biol 599(117-23 (2007).
K. S. Echtay, D. Roussel, J. St-Pierre, M. B. Jekabsons, S. Cadenas, J. A. Stuart, J. A. Harper, S. J. Roebuck, A. Morrison, S. Pickering, J. C. Clapham, and M. D. Brand, Superoxide activates mitochondrial uncoupling proteins, Nature 415(6867), 96-9 (2002).
K. S. Echtay, T. C. Esteves, J. L. Pakay, M. B. Jekabsons, A. J. Lambert, M. Portero-Otin, R. Pamplona, A. J. Vidal-Puig, S. Wang, S. J. Roebuck, and M. D. Brand, A signalling role for 4-hydroxy-2-nonenal in regulation of mitochondrial uncoupling, Embo J 22(16), 4103-10 (2003).
F. Palm, D. G. Buerk, P. O. Carlsson, P. Hansell, and P. Liss, Reduced nitric oxide concentration in the renal cortex of streptozotocin-induced diabetic rats: effects on renal oxygenation and microcirculation, Diabetes 54(11), 3282-7 (2005).
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Friederich, M., Nordquist, L., Olerud, J., Johansson, M., Hansell, P., Palm, F. (2009). Identification and Distribution of Uncoupling Protein Isoforms in the Normal and Diabetic Rat Kidney. In: Liss, P., Hansell, P., Bruley, D.F., Harrison, D.K. (eds) Oxygen Transport to Tissue XXX. Advances in Experimental Medicine and Biology, vol 645. Springer, Boston, MA. https://doi.org/10.1007/978-0-387-85998-9_32
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DOI: https://doi.org/10.1007/978-0-387-85998-9_32
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