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It is often assumed that pH(i) is spatially uniform within cells. A double-barreled microperfusion system was used to apply solutions of weak acid (acetic acid, CO(2)) or base (ammonia) to localized regions of an isolated ventricular myocyte (guinea pig). A stable, longitudinal pH(i) gradient (up to 1 pH(i) unit) was observed (using confocal imaging of SNARF-1 fluorescence). Changing the fractional exposure of the cell to weak acid/base altered the gradient, as did changing the concentration and type of weak acid/base applied. A diffusion-reaction computational model accurately simulated this behavior of pH(i). The model assumes that H(i)(+) movement occurs via diffusive shuttling on mobile buffers, with little free H(+) diffusion. The average diffusion constant for mobile buffer was estimated as 33 x 10(-7) cm(2)/s, consistent with an apparent H(i)(+) diffusion coefficient, D(H)(app), of 14.4 x 10(-7) cm(2)/s (at pH(i) 7.07), a value two orders of magnitude lower than for H(+) ions in water but similar to that estimated recently from local acid injection via a cell-attached glass micropipette. We conclude that, because H(i)(+) mobility is so low, an extracellular concentration gradient of permeant weak acid readily induces pH(i) nonuniformity. Similar concentration gradients for weak acid (e.g., CO(2)) occur across border zones during regional myocardial ischemia, raising the possibility of steep pH(i) gradients within the heart under some pathophysiological conditions.

Original publication

DOI

10.1529/biophysj.104.051391

Type

Journal article

Journal

Biophys J

Publication Date

04/2005

Volume

88

Pages

3018 - 3037

Keywords

Acetates, Ammonia, Animals, Biophysics, Buffers, Carbon, Carbon Dioxide, Cell Size, Cytoplasm, Diffusion, Guinea Pigs, HEPES, Heart Ventricles, Hydrogen-Ion Concentration, Microscopy, Confocal, Microscopy, Fluorescence, Models, Biological, Models, Cardiovascular, Models, Molecular, Models, Statistical, Myocytes, Cardiac, Perfusion, Proton-Motive Force, Protons, Quaternary Ammonium Compounds, Sarcolemma