The cytosolic free calcium in anti-μ-stimulated human B cells is derived partly from extracellular medium and partly from intracellular stores

The inositol phospholipid metabolism and the increase in cytosolic free Ca²⁺ concentration ([Ca²⁺]_i) into the cell are recognized as two important events in the anti-μ-induced B cell activation. The anti-μ stimulation caused the [³H]inositol incorporation and also a rapid increase in [Ca²⁺]_i from 85 nM to 285 nM. This signal returned to baseline a few minutes after stimulation. By using the fluorescent indicator quin-2 we demonstrated that this [Ca²⁺]_i uptake was derived part from extracellular medium and part from intracellular stores. Both EGTA (a calcium chelator) and TMB.8 (a drug which interferes with Ca²⁺ sequestration by smooth endoplasmic retiulum) partially suppressed the intracellular Ca²⁺ uptake and were fully inhibitory when added together. The role of Ca²⁺ from intracellular stores may also be evidenced in calcium-free experiments, or in permeabilized experiments using exogenous inositol 1,4,5-trisphosphate (IP₃, the putative mobilizer of intracellular Ca²⁺). Preventing the increase in [Ca²⁺]_i also prevents the apparition of early activation markers. These results are consistent with the hypothesis that the Ca²⁺ increase in B cells stimulated by anti-μ is caused by the generation of IP₃ during the phosphatidyl-inositol metabolism and also by the entry of extracellular Ca²⁺ through the plasma membrane.