Calcium imaging in intact mouse acinar cells in acute pancreas tissue slices

• Published in: PloS one 2022-06, Vol.17 (6), p.e0268644-e0268644
• Main Authors: Marolt, Urška, Paradiž Leitgeb, Eva, Pohorec, Viljem, Lipovšek, Saška, Venglovecz, Viktória, Gál, Eleonóra, Ébert, Attila, Menyhárt, István, Potrč, Stojan, Gosak, Marko, Dolenšek, Jurij, Stožer, Andraž
• Format: Article
• Language: English
• Subjects: Acetylcholine; Acinar cells; Biology and Life Sciences; Calcium; Calcium imaging; Calcium ions; Calcium signalling; Confocal microscopy; Development and progression; Electron microscopy; Enzymes; Glucose; Health aspects; Heterogeneity; Imaging; Immunofluorescence; Kinases; Medicine and Health Sciences; Methods; Microscopy; Morphology; Pancreas; Pancreatic diseases; Pancreatitis; Pathophysiology; Physical characteristics; Physiological aspects; Physiology; Research and Analysis Methods; Stains & staining; Stimulation; Tissues; Transmission electron microscopy
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0268644

The physiology and pathophysiology of the exocrine pancreas are in close connection to changes in intra-cellular Ca2+ concentration. Most of our knowledge is based on in vitro experiments on acinar cells or acini enzymatically isolated from their surroundings, which can alter their structure, physiology, and limit our understanding. Due to these limitations, the acute pancreas tissue slice technique was introduced almost two decades ago as a complementary approach to assess the morphology and physiology of both the endocrine and exocrine pancreas in a more conserved in situ setting. In this study, we extend previous work to functional multicellular calcium imaging on acinar cells in tissue slices. The viability and morphological characteristics of acinar cells within the tissue slice were assessed using the LIVE/DEAD assay, transmission electron microscopy, and immunofluorescence imaging. The main aim of our study was to characterize the responses of acinar cells to stimulation with acetylcholine and compare them with responses to cerulein in pancreatic tissue slices, with special emphasis on inter-cellular and inter-acinar heterogeneity and coupling. To this end, calcium imaging was performed employing confocal microscopy during stimulation with a wide range of acetylcholine concentrations and selected concentrations of cerulein. We show that various calcium oscillation parameters depend monotonically on the stimulus concentration and that the activity is rather well synchronized within acini, but not between acini. The acute pancreas tissue slice represents a viable and reliable experimental approach for the evaluation of both intra- and inter-cellular signaling characteristics of acinar cell calcium dynamics. It can be utilized to assess many cells simultaneously with a high spatiotemporal resolution, thus providing an efficient and high-yield platform for future studies of normal acinar cell biology, pathophysiology, and screening pharmacological substances.