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J Physiol Volume 541, Number 1, 139-158, May 15, 2002 DOI: 10.1113/jphysiol.2001.013267
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Journal of Physiology (2002), 541.1, pp. 139-158
© Copyright 2002 The Physiological Society
DOI: 10.1113/jphysiol.2001.013267

Intrinsic H+ ion mobility in the rabbit ventricular myocyte

R. D. Vaughan-Jones *, B. E. Peercy †, J. P. Keener † and K. W. Spitzer ‡

* University Laboratory of Physiology, Parks Road, Oxford OX1 3PT, UK, ‡ Nora Eccles Harrison Cardiovascular Research and Training Institute and † Department of Mathematics, University of Utah, Salt Lake City, UT, USA

The intrinsic mobility of intracellular H+ ions was investigated by confocally imaging the longitudinal movement of acid inside rabbit ventricular myocytes loaded with the acetoxymethyl ester (AM) form of carboxy-seminaphthorhodafluor-1 (carboxy-SNARF-1). Acid was diffused into one end of the cell through a patch pipette filled with an isotonic KCl solution of pH 3.0. Intracellular H+ mobility was low, acid taking 20-30 s to move 40 µm down the cell. Inhibiting sarcolemmal Na+-H+ exchange with 1 mM amiloride had no effect on this time delay. Net H+i movement was associated with a longitudinal intracellular pH (pHi) gradient of up to 0.4 pH units. H+i movement could be modelled using the equations for diffusion, assuming an apparent diffusion coefficient for H+ ions (DHapp) of 3.78 times 10-7 cm2 s-1, a value more than 300-fold lower than the H+ diffusion coefficient in a dilute, unbuffered solution. Measurement of the intracellular concentration of SNARF (~400 µM) and its intracellular diffusion coefficient (0.9 times 10-7 cm2 s-1) indicated that the fluorophore itself exerted an insignificant effect (between 0.6 and 3.3 %) on the longitudinal movement of H+ equivalents inside the cell. The longitudinal movement of intracellular H+ is discussed in terms of a diffusive shuttling of H+ equivalents on high capacity mobile buffers which comprise about half (~11 mM) of the total intrinsic buffering capacity within the myocyte (the other half being fixed buffer sites on low mobility, intracellular proteins). Intrinsic H+i mobility is consistent with an average diffusion coefficient for the intracellular mobile buffers (Dmob) of ~9 times 10-7 cm2 s-1.



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