Characterization of a large conductance non-selective anion channel in B lymphocytes

Studies on ion channel currents in freshly isolated murine B lymphocytes with the patch clamp technique revealed the presence of a non-selective anion channel of large conductance in inside-out (i/o) patches. This channel is characterized here according to its unitary conductance, ion selectivity, regulatory factors, distribution and kinetic behaviour. With a unitary conductance of 348±4.4 pS in a normal physiological ion gradient, it exhibited an indiscriminate selectivity to cations (Na + and K +). Selectivity to chloride over sodium was established by substitution of high concentrations of NaCl (450 mM) in the bath (i/o patches), resulting in a selectivity ratio (P Cl/P Na) of 33. Selectivity to chloride over potassium was confirmed in a similar manner by substitution of TEA-Cl for KCl, yielding a selectivity ratio (P Cl/P K)>80. Conductance of aspartate through the channel demonstrated the non-selective nature of this anion channel. Voltage proved to be a regulatory factor but other influences on channel activity were also present, including the configuration of the patch (channel is inactive in cell attached patches), and the enhancement of activity at negative membrane voltages by previous pulsing. Intracellular levels of calcium (i/o patches) did not appear to control channel conductances or regulate kinetic activity. Kinetic behaviour of this channel was complex, with periods of bursting and flickering activity interspersed with prolonged closed/open intervals. Multiple subconductance states were also present. The complex properties and behaviour of this channel suggest a possible role in signal transduction in B cell activation.