A novel approach to in situ characterization of pancreatic beta-cells

The tissue-slice technique has enabled major insights into neural and neuroendocrine physiology. Our aim was to adapt this technique to study the function of the endocrine pancreas. The preparation combines an in situ approach, as in gland perfusion, with a resolution characteristic of electrophysiological studies on single cells. The membrane potential in beta-cells in the slices recorded using the whole-cell patch-clamp was close to the calculated reversal potential for K+. With sufficient ATP in the recording pipette the beta-cells depolarized rapidly on exposure to an increased glucose concentration or stimulation with tolbutamide. The cells preserved bursting and spiking capacity for tens of minutes despite the whole-cell dialysis. In addition, the voltage clamp was used to monitor the changes in the membrane capacitance and to allow correlation of the electrical activity and the cytosolic calcium changes. The pancreatic tissue slice preparation is a novel method for studying the function of the beta- and other pancreatic endocrine and exocrine cells under near-physiological conditions.