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Am J Physiol Renal Physiol 273: F289-F299, 1997;
0363-6127/97 $5.00
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AJP - Renal Physiology, Vol 273, Issue 2 289-F299, Copyright © 1997 by American Physiological Society

ARTICLES

Monoclonal antibodies for high-resolution localization of NHE3 in adult and neonatal rat kidney

D. Biemesderfer, P. A. Rutherford, T. Nagy, J. H. Pizzonia, A. K. Abu-Alfa and P. S. Aronson
Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut 06520, USA.

Previous immunochemical studies have shown that NHE3 is an apical Na+/H+ exchanger in some renal epithelia. The purpose of the present study was to develop high-affinity, isoform-specific monoclonal antibodies (MAbs) that would be useful for carrying out high-resolution immunocytochemical studies of NHE3 in the adult and neonatal mammalian kidney. Three MAbs were developed to a fusion protein containing amino acids 702-832 of rabbit NHE3. Specificity was established by immunoblotting membranes from NHE-deficient LAP cells that had been transfected with either NHE1,-2, -3, or -4. With the use of high-resolution immunocytochemical techniques, NHE3 was found in vesicles in the apical cytoplasm of proximal tubule cells, as well as in the apical plasma membrane of the proximal tubule, and in both the thin and thick limbs of the loop of Henle. When localized in the 1-day-old rat kidney, NHE3 was first detected in the late stages of the S-shaped body. In later stages of nephron development, the pattern of NHE3 staining was similar to that seen in the adult. This study demonstrates 1) the specificity of three MAbs for Na+/H+ exchanger isoform NHE3; 2) NHE3 is present in an intracellular vesicular compartment in cells of the proximal tubule, consistent with possible regulation by membrane recycling; and 3) NHE3 is expressed on the apical membrane in early stages of the developing nephron.


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 J. Am. Soc. Nephrol.Home page
G.-H. KIM, C. ECELBARGER, M. A. KNEPPER, and R. K. PACKER
Regulation of Thick Ascending Limb Ion Transporter Abundance in Response to Altered Acid/Base Intake
J. Am. Soc. Nephrol., May 1, 1999; 10(5): 935 - 942.
[Abstract] [Full Text]

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 Am. J. Physiol. Renal Physiol.Home page
Y. Zhang, J. M. Norian, C. E. Magyar, N.-H. Holstein-Rathlou, A. K. Mircheff, and A. A. McDonough
In vivo PTH provokes apical NHE3 and NaPi2 redistribution and Na-K-ATPase inhibition
Am J Physiol Renal Physiol, May 1, 1999; 276(5): F711 - F719.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Renal Physiol.Home page
D. W. Good, T. George, and D. H. Wang
Angiotensin II inhibits HCO-3 absorption via a cytochrome P-450-dependent pathway in MTAL
Am J Physiol Renal Physiol, May 1, 1999; 276(5): F726 - F736.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
L. Fan, M. R. Wiederkehr, R. Collazo, H. Wang, L. A. Crowder, and O. W. Moe
Dual Mechanisms of Regulation of Na/H Exchanger NHE-3 by Parathyroid Hormone in Rat Kidney
J. Biol. Chem., April 16, 1999; 274(16): 11289 - 11295.
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 J. Biol. Chem.Home page
B. A. Watts III, T. George, and D. W. Good
Nerve Growth Factor Inhibits HCO3- Absorption in Renal Thick Ascending Limb through Inhibition of Basolateral Membrane Na+/H+ Exchange
J. Biol. Chem., March 19, 1999; 274(12): 7841 - 7847.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
H. Zhao, M. R. Wiederkehr, L. Fan, R. L. Collazo, L. A. Crowder, and O. W. Moe
Acute Inhibition of Na/H Exchanger NHE-3 by cAMP. ROLE OF PROTEIN KINASE A AND NHE-3 PHOSPHOSERINES 552 AND 605
J. Biol. Chem., February 12, 1999; 274(7): 3978 - 3987.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
M. Ikuma, M. Kashgarian, H. J. Binder, and V. M. Rajendran
Differential regulation of NHE isoforms by sodium depletion in proximal and distal segments of rat colon
Am J Physiol Gastrointest Liver Physiol, February 1, 1999; 276(2): G539 - G549.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Renal Physiol.Home page
B. M. Schmitt, D. Biemesderfer, M. F. Romero, E. L. Boulpaep, and W. F. Boron
Immunolocalization of the electrogenic Na+-HCO-3 cotransporter in mammalian and amphibian kidney
Am J Physiol Renal Physiol, January 1, 1999; 276(1): F27 - F38.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Renal Physiol.Home page
K.-P. Yip, C.-M. Tse, A. A. McDonough, and D. J. Marsh
Redistribution of Na+/H+ exchanger isoform NHE3 in proximal tubules induced by acute and chronic hypertension
Am J Physiol Renal Physiol, October 1, 1998; 275(4): F565 - F575.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Kurashima, E. Z. Szabo, G. Lukacs, J. Orlowski, and S. Grinstein
Endosomal Recycling of the Na+/H+ Exchanger NHE3 Isoform Is Regulated by the Phosphatidylinositol 3-Kinase Pathway
J. Biol. Chem., August 14, 1998; 273(33): 20828 - 20836.
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 J. Biol. Chem.Home page
D. Biemesderfer, B. DeGray, and P. S. Aronson
Membrane Topology of NHE3. EPITOPES WITHIN THE CARBOXYL-TERMINAL HYDROPHILIC DOMAIN ARE EXOPLASMIC
J. Biol. Chem., May 15, 1998; 273(20): 12391 - 12396.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Cell Physiol.Home page
Y. Zhang, C. E. Magyar, J. M. Norian, N.-H. Holstein-Rathlou, A. K. Mircheff, and A. A. McDonough
Reversible effects of acute hypertension on proximal tubule sodium transporters
Am J Physiol Cell Physiol, April 1, 1998; 274(4): C1090 - C1100.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
R. Collazo, L. Fan, M. C. Hu, H. Zhao, M. R. Wiederkehr, and O. W. Moe
Acute Regulation of Na+/H+ Exchanger NHE3 by Parathyroid Hormone via NHE3 Phosphorylation and Dynamin-dependent Endocytosis
J. Biol. Chem., October 6, 2000; 275(41): 31601 - 31608.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Szaszi, K. Kurashima, A. Kapus, A. Paulsen, K. Kaibuchi, S. Grinstein, and J. Orlowski
RhoA and Rho Kinase Regulate the Epithelial Na+/H+ Exchanger NHE3. ROLE OF MYOSIN LIGHT CHAIN PHOSPHORYLATION
J. Biol. Chem., September 8, 2000; 275(37): 28599 - 28606.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
D. Biemesderfer, B. DeGray, and P. S. Aronson
Active (9.6 S) and Inactive (21 S) Oligomers of NHE3 in Microdomains of the Renal Brush Border
J. Biol. Chem., March 23, 2001; 276(13): 10161 - 10167.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
M. C. Hu, L. Fan, L. A. Crowder, Z. Karim-Jimenez, H. Murer, and O. W. Moe
Dopamine Acutely Stimulates Na+/H+ Exchanger (NHE3) Endocytosis via Clathrin-coated Vesicles. DEPENDENCE ON PROTEIN KINASE A-MEDIATED NHE3 PHOSPHORYLATION
J. Biol. Chem., July 13, 2001; 276(29): 26906 - 26915.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Szaszi, K. Kurashima, K. Kaibuchi, S. Grinstein, and J. Orlowski
Role of the Cytoskeleton in Mediating cAMP-dependent Protein Kinase Inhibition of the Epithelial Na+/H+ Exchanger NHE3
J. Biol. Chem., October 26, 2001; 276(44): 40761 - 40768.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S. Breton, T. Wiederhold, V. Marshansky, N. N. Nsumu, V. Ramesh, and D. Brown
The B1 Subunit of the H+ATPase Is a PDZ Domain-binding Protein. COLOCALIZATION WITH NHE-RF IN RENAL B-INTERCALATED CELLS
J. Biol. Chem., June 9, 2000; 275(24): 18219 - 18224.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Renal Physiol.Home page
I. Sabolic, M. Ljubojevic, C. M. Herak-Kramberger, and D. Brown
Cd-MT causes endocytosis of brush-border transporters in rat renal proximal tubules
Am J Physiol Renal Physiol, December 1, 2002; 283(6): F1389 - F1402.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Renal Physiol.Home page
D. Biemesderfer, S. A. Mentone, M. Mooseker, and T. Hasson
Expression of myosin VI within the early endocytic pathway in adult and developing proximal tubules
Am J Physiol Renal Physiol, May 1, 2002; 282(5): F785 - F794.
[Abstract] [Full Text] [PDF]


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