Characterisation of calcium signalling in DT40 chicken B-cells

The chicken DT40 pre-B-cell line is becoming a potent experimental tool in the elucidation of higher organism cellular functions due to its unique genetic tractability. While several publications have described the effects of disruption of a range of genes in DT40 cells on calcium signalling, there has been no general overview of Ca2+ responses in wild-type cells. Here, we present experimental data comparing and contrasting the calcium responses to a range of agonists, such as alphaIgM, H2O2 and thapsigargin, applied singly or consecutively in the presence or absence of extracellular calcium. Briefly, we show that calcium release is from thapsigargin-sensitive and also -insensitive stores. This release results in, or is concomitant with, calcium entry across the plasma membrane through store-operated, receptor-operated and possibly L-type like Ca2+ channels. The agonists activate these pathways differentially producing a wide range of different sized and shaped Ca2+ signals. Furthermore, we report that Ca2+ responses in DT40 cells are dependent on the growth conditions. The presence of 1% chicken serum in the growth medium increased amplitudes of calcium responses and enhanced the sustained phase of the alphaIgM response, while 10 microM beta-mercaptoethanol in the medium (not, however, present during calcium measurements) resulted in more transient H2O2 responses and larger amplitude alphaIgM responses while failing to affect thapsigargin responses. The possible causes of these effects and their importance in comparing data from different studies on DT40 cells is discussed.