title: Frog striated muscle is permeable to hydroxide and buffer anions.
abstract: Hydroxide, bicarbonate and buffer anion permeabilities in semitendinosus muscle fibers of Rana pipiens were measured. In all experiments, the fibers were initially equilibrated in isotonic, high K2SO4 solutions at pHo = 7.2 buffered with phosphate. Two different methods were used to estimate permeabilities: (i) membrane potential changes were recorded in response to changes in external ion concentrations, and (ii) intracellular pH changes were recorded in response to changes in external concentrations of ions that alter intracellular pH. Constant field equations were used to calculate relative or absolute permeabilities. In the first method, to increase the size of the membrane potential change produced by a sudden change in anion entry, external K+ was replaced by Cs+ prior to changes of the anion under study. At constant external Cs+ activity, a hyperpolarization results from increasing external pH from 7.2 to 10.0 or higher, using either CAPS (3-[cyclohexylamino]-1-propanesulfonic acid) or CHES (2-[N-cyclohexylamino]-ethanesulfonic acid) as buffer. For each buffer, the protonated form is a zwitterion of zero net charge and the nonprotonated form is an anion. Using reported values of H+ permeability, calculations show that the reduction in [H+]o cannot account for the hyperpolarizations produced by alkaline solutions. Membrane hyperpolarization increases with increasing total external buffer concentration at constant external pH, and with increasing external pH at constant external buffer anion concentration. Taken together, these observations indicate that both OH- and buffer anions permeate the surface membrane. The following relative permeabilities were obtained at pHo = 10.0 +/- 0.3: (POH/PK) = 890 +/- 150, (PCAPS/PK) = 12 +/- 2, (PCHES/PK) = 5.3 +/- 0.9, and (PNO3/PK) = 4.7 +/- 0.5. PNO3/PK was independent of pHo up to 10.75. At pHo = 9.6, (PHCO3/PK) = 0.49 +/- 0.03; at pHo = 8.9, (PCl/PK) = 18 +/- 2 and at pHo = 7.1, (PHEPES/PK) = 20 +/- 2. In the second method, on increasing external pH from 7.2 to 10.0, using 2.5 mM CAPS (total buffer concentration), the internal pH increases linearly with time over the next 10 min. This alkalinization is due to the entry of OH- and the absorption of internal H+ by entering CAPS- anion. The rate of CAPS- entry was determined in experiments in which the external CAPS concentration was increased at constant external pH. Such increases invariably produced an increase in the rate of internal alkalinization, which was reversed when the CAPS concentration was reduced to its initial value.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Physiology, School of Medicine and Dentistry, University of Rochester, New York 14642.
authors.forename: R A
authors.initials: RA
authors.lastname: Venosa
authors.forename: B A
authors.initials: BA
authors.lastname: Kotsias
authors.forename: P
authors.initials: P
authors.lastname: Horowicz
mesh_terms: D000818:Animals; D000838:Anions / Q000493:pharmacokinetics*; D001639:Bicarbonates / Q000493:pharmacokinetics; D002021:Buffers; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D002586:Cesium / Q000493:pharmacokinetics; D006531:HEPES / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006878:Hydroxides / Q000493:pharmacokinetics*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009132:Muscles / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D009566:Nitrates / Q000493:pharmacokinetics; D010539:Permeability / Q000187:drug effects; D011188:Potassium / Q000493:pharmacokinetics; D011894:Rana pipiens / Q000502:physiology*
chemical_list: D000838:Anions; D001639:Bicarbonates; D002021:Buffers; D006878:Hydroxides; D009566:Nitrates; D002586:Cesium; C031356:hydroxide ion; D011188:Potassium; D006531:HEPES