Glucose-induced [Ca2+]i oscillations in β cells are composed of trains of spikes within a subplasmalemmal microdomain

Abstract

Electrical activity and oscillations of cytosolic Ca²⁺ concentrations ([Ca²⁺]_i) that trigger insulin release in response to glucose are key functions of pancreatic β cells. Although oscillatory Ca²⁺ signals have been intensively studied in β cells, their lower frequency did not match that of electrical activity. In addition, the measured peak [Ca²⁺]_i did not reach levels that are typically required by synaptotagmins to elicit the release of insulin-containing vesicles in live-cell experiments. We therefore sought to resolve the Ca²⁺ dynamics in the subplasmalemmal microdomain that is critical for triggering fast exocytosis. Applying total internal reflection fluorescence (TIRF) microscopy in insulin-producing INS-1E and primary mouse β cells, we resolved extraordinary fast trains of Ca²⁺ spiking (frequency > 3 s^-1) in response to glucose exposure. Using a low-affinity [Ca²⁺]_i indicator dye, we provide experimental evidence that Ca²⁺ spikes reach low micromolar apparent concentrations in the vicinity of the plasma membrane_. Analysis of Ca²⁺ spikes evoked by repeated depolarization for 10 ms closely matched the Ca²⁺ dynamics observed upon glucose application. To our knowledge, this is the first study that experimentally demonstrates Ca²⁺ spikes in β cells with velocities that resemble those of bursting or continuously appearing trains of action potentials (APs) in non-patched cells.

Keywords: Calcium oscillation; Microdomains; Pancreatic beta cells; Total internal reflection microscopy.