Effects of 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate (TMB8) on rat atrial muscle.

Rat atria loaded in vitro with the dye INDO-1 produced fluorescence signals indicative of changes in cytoplasmic calcium ion concentration ([Ca2+]c). Such atria showed systolic/diastolic fluctuations in fluorescence indicative of a systolic rise and a diastolic fall in [Ca2+] while being superfused with a solution containing a normal Ca2+ concentration. Some atria were then exposed to a low [Ca2+] in the superfusate. This caused negative inotropism and fluorescence changes indicative of a decline in [Ca2+]c. Both of these responses were reversed by adding 8-(N,N-diethylamino)octyl 3,4,5-trimethoxybenzoate (TMB8; 2 microM) to the superfusate. Some atria were exposed instead either to a low [K+] in the superfusate or to an ouabain-containing superfusate. These atria developed a contracture, associated with fluorescence changes indicative of a rise in [Ca2+]c. The addition of TMB8 (2 microM) now relaxed the contracture, and this was associated with fluorescence changes indicative of a decline in [Ca2+]c. Atria that were exposed for 15 min to a low [Na+] in the superfusate developed a period of positive inotropism, followed by a brief period of negative inotropism on return to the normal superfusate. The period of positive inotropism was associated with fluorescence changes indicative of a rise in [Ca2+]c and the period of negative inotropism with a decline in [Ca2+]c to below baseline levels. All of these responses were less marked in atria exposed throughout to superfusates containing TMB8 (2 microM). Some atria were loaded with the dye SNARF-1. This emits fluorescence signals indicative of changes in cytoplasmic pH (pHc). These atria showed no systolic/diastolic fluctuation of fluorescence, but when superfused with a bicarbonate-free solution they displayed a change in fluorescence indicative of a decline in pHc in response to the addition of either ouabain or TMB8. Similarities were found between the effects produced by TMB8 and those produced by amiloride or dichlorobenzyl amiloride, suggesting that all three agents inhibit plasmalemmal Na+/Ca2+ and Na+/H+ exchange.