Abstract

The mechanism of stimulation of Ca2+ entry into hepatocytes by adenosine was investigated. When Fura-2-loaded hepatocytes were suspended in a nominally Ca2+-free buffer, adenosine produced only a small transient increase in the cytosolic free Ca2+ concentration ([Ca2+](i)). However, on restoration of an extracellular Ca2+ concentration of 1.3 mM, a rapid increase in [Ca2+](i), occurred, which indicates activation of a Ca2+-influx pathway. Adenosine augmented the rare of Ca2+ influx triggered by maximally effective concentrations of thapsigargin or cAMP, but was without effect on the rate of Ca2+ entry that resulted from phospholipase-C-linked-receptor activation by maximally effective concentrations of vasopressin or ATP. However, in contrast to vasopression and ATP, adenosine did not stimulate Mn2+ entry. The rate of Mn2+ influx after stimulation of the hepatocytes with vasopressin was not increased by adenosine treatment. The stimulation of hepatocytes with adenosine did not result in significant accumulation of inositol phosphates or cAMP. Furthermore, the rate of adenosine-induced Ca2+ entry in hepatocytes was only slightly reduced in the presence of the P-1 purinoceptor antagonist 8-phenyltheophylline. In contrast, the receptor-mediated-Ca2+-entry antagonist SK 96365 nearly completely blocked the Ca2+-entry response without any effect on internal-Ca2+-pool mobilisation by adenosine. it is concluded that adenosine activates the internal-pool-regulated pathway of Ca2+ entry and an additional pathway that appears comparable to the previously reported receptor-dependent pathway, except that Mn2+ entry is nor stimulated.