title: Direct evidence for apical Na+:2Cl-:K+ cotransport in macula densa cells.
abstract: Previous studies by our laboratory indicate that increases in apical NaCl concentration ([NaCl]) depolarize macula densa (MD) cells, although the mechanism for apical NaCl transport was not identified. To determine the pathway for MD apical NaCl transport, we utilized microdissected cortical thick ascending limbs (CTAL) with attached glomeruli and conventional microelectrode techniques. Addition of 50 microM furosemide in the presence of 150 mM NaCl produced a variable hyperpolarization of basolateral membrane voltage (delta Vbl, -14 +/- 8.2 mV, NS P = 0.15, n = 6) and completely blocked the expected repolarization on reducing luminal [NaCl] from 150 to 25 mM. Addition of furosemide in the presence of 25 mM NaCl depolarized Vbl by 22 +/- 6.8 mV (P less than 0.05, n = 6) indicating that the direction of the NaCl transport can be reversed in low luminal [NaCl]. In other studies, luminal concentration of Na or Cl was increased from 25 to 150 mM. Increased [Na] produced a 6.9 +/- 1.2 mV (n = 9) depolarization, whereas Cl addition depolarized Vbl by 8.2 +/- 1.7 mV (n = 5), suggesting that both ions are involved in the NaCl-induced MD depolarization. Removal of K from the luminal perfusate elicited a hyperpolarization of -14 +/- 2.9 mV (n = 9). These results are all consistent with the existence of an apical Na+:2Cl-:K+ transporter that would result in NaCl reabsorption in the presence of 150 mM luminal NaCl but would produce NaCl secretion at low luminal NaCl concentrations.
authors.affiliation: Membrane Transport Research Group, University of Montreal, Quebec, Canada.
authors.forename: J Y
authors.initials: JY
authors.lastname: Lapointe
authors.forename: P D
authors.initials: PD
authors.lastname: Bell
authors.forename: J
authors.initials: J
authors.lastname: Cardinal
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D005260:Female; D005665:Furosemide / Q000494:pharmacology; D007425:Intracellular Membranes / Q000502:physiology; D007684:Kidney Tubules / Q000378:metabolism*; D008138:Loop of Henle / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters
chemical_list: D002352:Carrier Proteins; D002712:Chlorides; D028021:Sodium-Potassium-Chloride Symporters; D012965:Sodium Chloride; D005665:Furosemide; D012964:Sodium; D011188:Potassium