Secretagogue-induced oscillations of cytoplasmic Ca2+ in single β and α-cells obtained from pancreatic islets by fluorescence-activated cell sorting

Abstract Pancreatic islet cells from normal adult rats were purified by autofluorescence-activated cell sorting (FACS). A purification of islet β-cells (>90%) and α-cells (>70%) was obtained after FACS separation. Dual-wavelength microfluorimetry was employed for measurements of cytoplasmic Ca 2+ concentration ( Ca 2+ i ) in single β- and α-cells. In FACS-purified β-cells, 11.8 mM glucose induced oscillations of Ca 2+ i with a frequency of 1 cycle per 2–4 min, whereas 20 mM glucose induced a rapid rise in Ca 2+ i followed by a sustained elevation in Ca 2+ i . In contrast, α-cells exhibited a decrease in Ca 2+ i following exposure to 20 mM glucose, and subsequent exposure to 20 mM arginine induced a large increase in Ca 2+ i with periodic fluctuations. FACS purification of a heterogeneous islet-cell population provides a unique model for characterizing secretagogue-induced changes in Ca 2+ i in single β- and α-cells.

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