title: Effects of dietary salt on renal Na+ transporter subcellular distribution, abundance, and phosphorylation status.
abstract: During high-salt (HS) diet the kidney increases urinary Na+ and volume excretion to match intake. We recently reported that HS provokes a redistribution of distal convoluted tubule Na+-Cl- cotransporter (NCC) from apical to subapical vesicles and decreases NCC abundance. This study aimed to test the hypothesis that the other renal Na+ transporters' abundance and or subcellular distribution is decreased by HS diet. Six-week-old Sprague-Dawley rats were fed a normal (NS) 0.4% NaCl diet or a HS 4% NaCl diet for 3 wk or overnight. Kidneys excised from anesthetized rats were fractionated on density gradients or analyzed by microscopy; transporters and associated regulators were detected with specific antibodies. Three-week HS doubled Na+/H+ exchanger (NHE)3 phosphorylation at serine 552 and provoked a redistribution of NHE3, dipeptidyl peptidase IV (DPPIV), myosin VI, Na+-Pi cotransporter (NaPi)-2, ANG II type 2 receptor (AT2R), aminopeptidase N (APN), Na+-K+-2Cl- cotransporter (NKCC2), epithelial Na+ channel (ENaC) beta-subunit, and Na+-K+-ATPase (NKA) alpha1- and beta1-subunits from low-density plasma membrane-enriched fractions to higher-density intracellular membrane-enriched fractions. NHE3, myosin VI, and AT2R retraction to the base of the microvilli (MV) during HS was evident by confocal microscopy. HS did not change abundance of NHE3, NKCC, or NKA alpha1- or beta1-subunits but increased ENaC-beta in high-density intracellular enriched membranes. Responses to HS were fully apparent after just 18 h. We propose that retraction of NHE3 to the base of the MV, driven by myosin VI and NHE3 phosphorylation and accompanied by redistribution of the NHE3 regulator DPPIV, contributes to a decrease in proximal tubule Na+ reabsorption during HS and that redistribution of transporters out of low-density plasma membrane-enriched fractions in the thick ascending limb of the loop of Henle and distal nephron may also contribute to the homeostatic natriuretic response to HS diet.
authors.affiliation: Department of Physiology and Biophysics, University of Southern California Keck School of Medicine, 1333 San Pablo St., MMR 626, Los Angeles, CA 90089-9142, USA.
authors.forename: Li E
authors.initials: LE
authors.lastname: Yang
authors.forename: Monica B
authors.initials: MB
authors.lastname: Sandberg
authors.forename: Argun D
authors.initials: AD
authors.lastname: Can
authors.forename: Kaarina
authors.initials: K
authors.lastname: Pihakaski-Maunsbach
authors.forename: Alicia A
authors.initials: AA
authors.lastname: McDonough
mesh_terms: D000818:Animals; D018826:CD13 Antigens / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism; D053503:Epithelial Sodium Channels / Q000378:metabolism; D007672:Kidney Cortex / Q000378:metabolism*; D007679:Kidney Medulla / Q000378:metabolism*; D008297:Male; D018995:Myosin Heavy Chains / Q000378:metabolism; D009318:Natriuresis / Q000502:physiology*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D044139:Receptor, Angiotensin, Type 2 / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D017673:Sodium Chloride, Dietary / Q000494:pharmacology*; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D064466:Solute Carrier Family 12, Member 1
chemical_list: D002352:Carrier Proteins; D053503:Epithelial Sodium Channels; D010750:Phosphoproteins; D044139:Receptor, Angiotensin, Type 2; C578716:Slc12a1 protein, rat; C000613387:Slc9a3 protein, rat; D017673:Sodium Chloride, Dietary; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; D028021:Sodium-Potassium-Chloride Symporters; D064466:Solute Carrier Family 12, Member 1; C089785:myosin VI; C093376:sodium-hydrogen exchanger regulatory factor; D012964:Sodium; D018826:CD13 Antigens; D018819:Dipeptidyl Peptidase 4; D018995:Myosin Heavy Chains; D000254:Sodium-Potassium-Exchanging ATPase