Effects of plasmalemmal V-ATPase activity on plasma membrane potential of resident alveolar macrophages.
The acid-base status and functional responses of alveolar macrophages (mphi) are influenced by the activity of plasmalemmal V-type H+-pump (V-ATPase), an electrogenic H+ extruder that provides a possible link between intracellular pH (pHi) and plasma membrane potential (Em). This study examined the relationships among Em, pHi, and plasmalemmal V-ATPase activity in resident alveolar mphi from rabbits. Em and pHi were measured using fluorescent probes. Em was -46 mV and pHi was 7.14 at an extracellular pH (pHo) of 7.4. The pHi declined progressively at lower pHo values. Decrements in pHo, also caused depolarization of the plasma membrane, independent of V-ATPase activity. The pH effects on Em were sensitive to external K+, and hence, probably involved pH-sensitive K+ conductance. H+ were not distributed at equilibrium across the plasma membrane. V-ATPase activity was a major determinant of the transmembrane H+ disequilibrium. Pump inhibition with bafilomycin A1 caused cytosolic acidification, due most likely to the retention of metabolically generated H+. V-ATPase inhibition also caused depolarization of the plasma membrane, but the effects were mediated indirectly via the accompanying pHi changes. V-ATPase activity was sensitive to Em. Em hyperpolarization (valinomycin-clamp) reduced V-ATPase activity, causing an acidic shift in baseline pHi under steady-state conditions and slowing pHi recovery from NH4Cl prepulse acid-loads. The findings indicate that a complex relationship exists among Em, pHi, and pHo that was partially mediated by plasmalemmal V-ATPase activity. This relationship could have important consequences for the expression of pH- and/or voltage-sensitive functions in alveolar mphi.
Heming
T A
TA
Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Texas Health Science Center, Houston, Texas 77030, USA. Thomas.Heming@uth.tmc.edu
Bidani
A
A
D000136:Acid-Base Equilibrium / Q000502:physiology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D007424:Intracellular Fluid / Q000737:chemistry; D016676:Macrophages, Alveolar / Q000201:enzymology / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology; D011817:Rabbits; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism / Q000502:physiology*
D025262:Vacuolar Proton-Translocating ATPases