Bumann J, Wurster B, Malchow D
J Cell Biol. 1984 Jan;98(1):173-8. doi: 10.1083/jcb.98.1.173.
We used a Ca++-sensitive electrode to measure changes in extracellular Ca++ concentration in cell suspensions of Dictyostelium discoideum during differentiation and attractant stimulation. The cells maintained an external level of 3-8 microM Ca++ until the beginning of aggregation and then started to take up Ca++. The attractants, folic acid, cyclic AMP, and cyclic GMP, induced a transient uptake of Ca++ by the cells. The response was detectable within 6 s and peaked at 30 s. Half-maximal uptake occurred at 5 nM cyclic AMP or 0.2 microM folic acid, respectively. The apparent rate of uptake amounted to 2 X 10(7) Ca++ per cell per min. Following uptake, Ca++ was released by the cells with a rate of 5 X 10(6) ions per cell per min. Specificity studies indicated that the induced uptake of Ca++ was mediated by cell surface receptors. The amount of accumulated Ca++ remained constant as long as a constant stimulus was provided. No apparent adaptation occurred. The cyclic AMP-induced uptake of Ca++ increased during differentiation and was dependent on the external Ca++ concentration. Saturation was found above 10 microM external Ca++. The time course and magnitude of the attractant-induced uptake of external Ca++ agree with a role of Ca++ during contraction. During development the extracellular Ca++ level oscillated with a period of 6-11 min. The change of the extracellular Ca++ concentration during one cycle would correspond to a 30-fold change of the cellular free Ca++ concentration.
我们使用钙离子敏感电极来测量盘基网柄菌细胞悬浮液在分化和趋化因子刺激过程中细胞外钙离子浓度的变化。细胞在聚集开始前维持细胞外钙离子水平为3 - 8微摩尔,然后开始摄取钙离子。趋化因子叶酸、环磷酸腺苷(cAMP)和环磷酸鸟苷(cGMP)可诱导细胞短暂摄取钙离子。该反应在6秒内可检测到,并在30秒时达到峰值。半最大摄取量分别出现在5纳摩尔环磷酸腺苷或0.2微摩尔叶酸时。摄取的表观速率为每分钟每个细胞2×10⁷个钙离子。摄取后,细胞以每分钟每个细胞5×10⁶个离子的速率释放钙离子。特异性研究表明,诱导的钙离子摄取是由细胞表面受体介导的。只要提供恒定刺激,积累的钙离子量就保持恒定。未出现明显的适应性变化。环磷酸腺苷诱导的钙离子摄取在分化过程中增加,并且依赖于细胞外钙离子浓度。在细胞外钙离子浓度高于10微摩尔时出现饱和。趋化因子诱导的细胞外钙离子摄取的时间进程和幅度与钙离子在收缩过程中的作用一致。在发育过程中,细胞外钙离子水平以6 - 11分钟的周期振荡。一个周期内细胞外钙离子浓度的变化相当于细胞内游离钙离子浓度30倍的变化。