Regulation of basolateral membrane potential after stimulation of Na+ transport in proximal tubules.
We have previously shown that stimulation of apical Na-coupled glucose and alanine transport produces a transient depolarization of basolateral membrane potential (Vbl) in rabbit proximal convoluted tubule (PCT, S1 segment). The present study is aimed at understanding the origin of the membrane repolarization following the initial effect of addition of luminal cotransported solutes. Luminal addition of 10-15 mM L-alanine produced a rapid and highly significant depolarization of Vbl (20.3 +/- 1.1 mV, n = 15) which was transient and associated with an increase in the fractional K+ conductance of the basolateral membrane (tK) from 8 to 29% (P less than 0.01, n = 6). Despite the significant increase in tK, the repolarization was only slightly reduced by the presence of basolateral Ba2+ (2 mM, n = 6) or quinine (0.5 mM, n = 5). The repolarization was greatly reduced in the presence of 0.1 mM 4-acetamino-4'isothiocyamostilbene-2,2'-disulfonic acid (SITS) and blunted by bicarbonate-free solutions. Intracellular pH (pHi) determined with the fluorescent dye 2',7'-bis-2-carboxyethyl-5(and -6)-carboxyfluorescein (BCECF), averaged 7.39 +/- 0.02 in control solution (n = 9) and increased to 7.50 +/- 0.03 in the first 15 sec after the luminal application of alanine. This was followed by a significant acidification averaging 0.16 +/- 0.01 pH unit in the next 3 min. In conclusion, we believe that, contrary to other leaky epithelia, rabbit PCT can regulate its basolateral membrane potential not only through an increase in K+ conductance but also through a cellular acidification reducing the basolateral HCO3- exit through the electrogenic Na-3(HCO3) cotransport mechanism.
Lapointe
J Y
JY
Membrane Transport Research Group, University of Montreal, Quebec, Canada.
Duplain
M
M
D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000494:pharmacology; D000818:Animals; D001464:Barium / Q000494:pharmacology; D001485:Basement Membrane / Q000378:metabolism; D001639:Bicarbonates / Q000494:pharmacology; D001693:Biological Transport, Active; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D010477:Perfusion; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D011803:Quinine / Q000494:pharmacology; D011817:Rabbits; D012964:Sodium / Q000378:metabolism*
D001639:Bicarbonates; D015221:Potassium Channels; D001464:Barium; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D012964:Sodium; D011803:Quinine