title: The plasma membrane calcium ATPase (PMCA) of neurones is electroneutral and exchanges 2 H+ for each Ca2+ or Ba2+ ion extruded.
abstract: The coupling between Ca2+ extrusion and H+ uptake by the ubiquitous plasma membrane calcium ATPase (PMCA) has not been measured in any neurone. I have investigated this with Ca2+- and pH-sensitive microelectrodes in large voltage-clamped snail neurones, which have no Na+-Ca2+ exchangers. The recovery of [Ca2+]i and surface pH after a brief depolarization or Ca2+ injection was not slowed by hyperpolarization to -90 mV from a holding potential of -50 mV, consistent with a 1 Ca2+ : 2 H+ coupling ratio. Since Ca2+ injections proved difficult to quantify, and Ca2+ currents through channels were obscured by K+ currents, Ba2+ was used as a substitute. When the cell was bathed in Ca2+-free Ba2+ Ringer solution, the K+ currents were blocked and large inward currents were revealed on depolarization. The Ca2+-sensitive microelectrodes were sensitive to intracellular Ba2+ as well as Ca2+. With equal depolarizations Ba2+ entry appeared larger than Ca2+ entry and generated similar but slower pH changes. Ba2+ extrusion was insensitive to hyperpolarization, blocked by eosin or high pH, and about 5 times slower than Ca2+ extrusion. The ratio of the pH change caused by the extrusion of unit charge of Ba2+ influx to that caused by unit charge of H+ injection was 0.85 +/- 0.08 (s.e.m., n = 8), corresponding to a Ba2+ : H+ ratio of 1 : 1.7. Both this ratio and the electroneutrality of the PMCA suggest that the Ca2+ : H+ ratio is 1 : 2, ensuring that after a Ca2+ influx [Ca2+]i recovery is not influenced by the membrane potential and maximizes the conversion of Ca2+ influxes into possible pH signals.
authors.affiliation: Physiological Laboratory, University of Cambridge, Downing Site, Cambridge, UK. rct26@cam.ac.uk
authors.forename: Roger C
authors.initials: RC
authors.lastname: Thomas
mesh_terms: D000818:Animals; D001464:Barium / Q000378:metabolism* / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004560:Electricity; D004801:Eosine Yellowish-(YS) / Q000494:pharmacology; D005724:Ganglia / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006372:Helix, Snails; D006851:Hydrochloric Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D017136:Ion Transport / Q000187:drug effects / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011522:Protons; D000254:Sodium-Potassium-Exchanging ATPase / Q000502:physiology
chemical_list: D011522:Protons; D001464:Barium; D004533:Egtazic Acid; D053499:Plasma Membrane Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase; D006851:Hydrochloric Acid; D002118:Calcium; D004801:Eosine Yellowish-(YS)