20.500.12556/DKUM-91652 Ultrafast multicellular calcium imaging of calcium spikes in mouse beta cells in tissue slices Background: The crucial steps in beta cell stimulus-secretion coupling upon stimulation with glucose are oscillatory changes in metabolism, membrane potential, intracellular calcium concentration, and exocytosis. The changes in membrane potential consist of bursts of spikes, with silent phases between them being dominated by membrane repolarization and absence of spikes. Assessing intra- and intercellular coupling at the multicellular level is possible with ever-increasing detail, but our current ability to simultaneously resolve spikes from many beta cells remains limited to double-impalement electrophysiological recordings. Methods: Since multicellular calcium imaging of spikes would enable a better understanding of coupling between changes in membrane potential and calcium concentration in beta cell collectives, we set out to design an appropriate methodological approach. Results: Combining the acute tissue slice method with ultrafast calcium imaging, we were able to resolve and quantify individual spikes within bursts at a temporal resolution of >150 Hz over prolonged periods, as well as describe their glucose-dependent properties. In addition, by simultaneous patch-clamp recordings we were able to show that calcium spikes closely follow membrane potential changes. Both bursts and spikes coordinate across islets in the form of intercellular waves, with bursts typically displaying global and spikes more local patterns. Conclusions: This method and the associated findings provide additional insight into the complex signaling within beta cell networks. Once extended to tissue from diabetic animals and human donors, this approach could help us better understand the mechanistic basis of diabetes and find new molecular targets. beta cell calcium imaging calcium oscillations calcium spikes physiology true false true John Wiley & Sons Ltd Angleški jezik Angleški jezik Znanstveno delo 2025-01-24 09:02:39 2025-01-24 09:16:32 2025-01-25 03:11:16 0000-00-00 00:00:00 2025 0 0 Str. 1-20 Št. 2 Letn. 241 Feb. 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 2024-11-29 2024-01-16 2025-02-01 612 222131203 10.1111/apha.14261 1748-1716 RAZ_Dolensek_Jurij_2025.pdf RAZ_Dolensek_Jurij_2025.pdf 1 DF3AE70739E4403A5B976D6665FE8394 174f8ff56b43e8e0f566af445fdcbe944583ae49279294f65bcc272649dbc248 c2ddfdcf-da2a-11ef-9445-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=188108 https://onlinelibrary.wiley.com/doi/10.1111/apha.14261 1 969325e8-da29-11ef-9445-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=188107 Medicinska fakulteta Fakulteta za naravoslovje in matematiko 0 0 0