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AJP - Renal Physiology, Vol 271, Issue 3 619-F628, Copyright © 1996 by American Physiological Society
ARTICLES |
C. A. Ecelbarger, J. Terris, J. R. Hoyer, S. Nielsen, J. B. Wade and M. A. Knepper
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda 20892, USA.
To investigate the role of the thick ascending limb (TAL) Na(+)-K(+)-2Cl- cotransporter in regulation of water excretion, we have prepared a peptide-derived polyclonal antibody based on the cloned cDNA sequence of the rat type 1 bumetanide-sensitive cotransporter, BSC-1 (also termed "NKCC-2"). Immunoblots revealed a single broad 161-kDa band in membrane fractions of rat renal outer medulla and cortex but not from rat colon or parotid gland. A similar protein was labeled in mouse kidney. Immunoperoxidase immunohistochemistry in rat kidney revealed labeling restricted to the medullary and cortical TAL segments. Because long-term regulation of urinary concentrating ability may depend on regulation of Na(+)-K(+)-2Cl- cotransporter abundance, we used immunoblotting to evaluate the effects of several in vivo factors on expression levels of BSC-1 protein in rat kidney outer medulla. Chronic oral saline loading with 0.16 M NaCl markedly increased BSC-1 abundance. However, long-term vasopressin infusion or thirsting of rats did not affect BSC-1 abundance. Chronic furosemide infusion caused a 9-kDa upward shift in apparent molecular mass and an apparent increase in expression level. These results support the previous identification of BSC-1 as the TAL Na(+)-K(+)-2Cl- transporter and demonstrate that the expression of this transporter is regulated.
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 S. M. Wall and M. P. Fischer Contribution of the Na+-K+-2Cl- Cotransporter (NKCC1) to Transepithelial Transport of H+, NH4+, K+, and Na+ in Rat Outer Medullary Collecting Duct J. Am. Soc. Nephrol., April 1, 2002; 13(4): 827 - 835. [Abstract] [Full Text] [PDF]
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C. A. Bertuccio, F. R. Ibarra, J. E. Toledo, E. E. Arrizurieta, and R. S. Martin Endogenous vasopressin regulates Na-K-ATPase and Na+-K+-Cl- cotransporter rbsc-1 in rat outer medulla Am J Physiol Renal Physiol, February 1, 2002; 282(2): F265 - F270. [Abstract] [Full Text] [PDF]
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A. D. Bradford, J. M. Terris, C. A. Ecelbarger, J. D. Klein, J. M. Sands, C.-L. Chou, and M. A. Knepper 97- and 117-kDa forms of collecting duct urea transporter UT-A1 are due to different states of glycosylation Am J Physiol Renal Physiol, July 1, 2001; 281(1): F133 - F143. [Abstract] [Full Text] [PDF]
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