2-Aminoethoxydiphenyl borate (2-APB) antagonises inositol 1,4,5-trisphosphate-induced calcium release, inhibits calcium pumps and has a use-dependent and slowly reversible action on store-operated calcium entry channels

The action of 2-aminoethoxydiphenyl borate (2-APB) on Ca2+ signalling in HeLa cells and cardiac myocytes was investigated. Consistent with other studies, we found that superfusion of cells with 2-APB rapidly inhibited inositol 1,4,5-trisphosphate (InsP(3))-mediated Ca2+ release and store-operated Ca2+ entry (SOC). In addition to abrogating hormone-evoked Ca2+ responses, 2-APB could antagonise Ca2+ signals evoked by a membrane permeant InsP(3) ester. 2-APB also slowed the recovery of intracellular Ca2+ signals consistent with an effect on Ca2+ ATPases. The inhibitory action of 2-APB on InsP(3) receptors (InsP(3)Rs), SOC channels and Ca2+ pumps persisted for several minutes after washout of the compound. Application of 2-APB to unstimulated cells had no effect on subsequent Ca2+ responses suggesting that it has a use-dependent action. Mitochondria in cells treated with 2-APB showed a rapid and slowly reversible swelling. 2-APB did not cause the mitochondria to depolarise, but it reduced the extent of mitochondrial calcium uptake. Although 2-APB has been demonstrated not to affect voltage-operated Ca2+ channels or ryanodine receptors, we found that it gave a concentration-dependent long-lasting inhibition of Ca2+ signalling in electrically-stimulated cardiac myocytes, where InsP(3)Rs and SOC channels do not play a significant role. Our data suggest that 2-APB has multiple cellular targets, a use-dependent action, is difficult to reverse and may affect Ca2+ signalling in cell types where InsP(3) and SOC are not active. (C) 2003 Elsevier Science Ltd. All rights reserved.