We previously demonstrated a transmural gradient in Na/K pump current (I P) and [Na+] i , with the highest maximum I P and lowest [Na+] i in epicardium. The present study examines the relationship between the transmural gradient in I P and Na/Ca exchange (NCX). Myocytes were isolated from canine left ventricle. Whole-cell patch clamp was used to measure current generated by NCX (I NCX) and inward background calcium current (I ibCa), defined as inward current through Ca2+ channels less outward current through Ca2+-ATPase. When resting myocytes from endocardium (Endo), midmyocardium (Mid) or epicardium (Epi) were studied in the same conditions, I NCX was the same and I ibCa was zero. Moreover, Western blots were consistent with NCX protein being uniform across the wall. However, the gradient in [Na+] i , with I ibCa = 0, should create a gradient in [Ca2+] i . To test this hypothesis, we measured resting [Ca2+] i using two methods, based on either transport or the Ca2+-sensitive dye Fura2. Both methods demonstrated a significant transmural gradient in resting [Ca2+] i , with Endo > Mid > Epi. This gradient was eliminated by exposing Epi to sufficient ouabain to partially inhibit Na/K pumps, thus increasing [Na+] i to values similar to those in Endo. These data support the existence of a transmural gradient for Ca2+ removal by NCX. This gradient is not due to differences in expression of NCX; rather, it is generated by a transmural gradient in [Na+] i , which is due to a transmural gradient in plasma membrane expression of the Na/K pump.
The Journal of Membrane Biology – Springer Journals
Published: Feb 4, 2010
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