Tolbutamide stimulation of pancreatic beta-cells involves both cell recruitment and increase in the individual Ca(2+)...

Individual pancreatic beta-cells are functionally heterogeneous. Their sensitivity to glucose is variable, so that the proportion of active cells increases with the glucose concentration (recruitment). We have investigated whether sulphonylureas also recruit beta-cells, by measuring cytoplasmic Ca(2+) ([Ca(2+)](i)) - the triggering signal of insulin secretion - in single cells and clusters of cells prepared from mouse islets. In 4 mM glucose, the threshold concentration of tolbutamide inducing a [Ca(2+)](i) rise was variable (5 - 50 microM). The proportion of responsive cells and clusters therefore increased with the tolbutamide concentration, to reach a maximum of 90% of the cells and 100% of the clusters. This recruitment occurred faster when the glucose concentration was increased from 4 to 5 mM (EC(50) of approximately 14 and approximately 4 microM tolbutamide respectively). Within responsive clusters little recruitment was observed; when a cluster was active, all or nearly all cells were active probably because of cell coupling. Thus, tolbutamide-induced [Ca(2+)](i) oscillations were synchronous in all cells of each cluster, whereas there was no synchrony between clusters or individual cells. Independently of cell recruitment, tolbutamide gradually augmented the magnitude of the [Ca(2+)](i) rise in single cells and clusters. This increase occurred over a broader range of concentrations than did recruitment (EC(50) of approximately 50 and 25 microM tolbutamide at 4 and 5 mM glucose respectively). Tolbutamide (10 microM) accelerated the recruitment of single cells and clusters brought about by increasing glucose concentrations (range of 3 - 7 mM instead of 4 - 10 mM glucose), and potentiated the amplification of the individual responses that glucose also produced. In conclusion, both metabolic (glucose) and pharmacologic (sulphonylurea) inhibition of K(+)-ATP channels recruits beta-cells to generate a [Ca(2+)](i) response. However, the response is not of an all-or-none type; it increases in amplitude with the concentration of either glucose or tolbutamide.