Ca2+and Mn2+Influx Through Receptor-Mediated Activation of Nonspecific Cation Channels in Mast Cells

Whole-cell patch-clamp recordings of membrane currents and Fura-2 measurements of free intracellular calcium concentration ([Ca2+]i) were used to study calcium influx through receptor-activated cation channels in rat peritoneal mast cells. Cation channels were activated by the secretagogue compound 48/80, whereas a possible concomitant Ca2+entry through pathways activated by depletion of calcium stores was blocked by dialyzing cells with heparin. Heparin effectively suppressed the transient Ca2+release induced by 48/80 and abrogated inositol 1,4,5-trisphosphate-induced calcium influx without affecting activation of 50-pS cation channels. There was a clear correlation between changes in [Ca2+]iand the activity of 50-pS channels. The changes in [Ca2+]iincreased with elevation of extracellular Ca2+. At the same time, inward currents through 50-pS channels were diminished as more Ca2+permeated. This effect was due to a decrease in slope conductance and a reduction in the open probability of the cation channels. In physiological solutions, 3.6% of the total current was carried by Ca2+. The cation channels were not only permeable to Ca2+but also to Mn2+, as evidenced by the quench of Fura-2 fluorescence. Mn2+current through 50-pS channels could not be resolved at the single-channel level. Our results suggest that 50-pS cation channels partially contribute to sustained increases of [Ca2+]iin mast cells following receptor activation.