Ca2+ signals in pancreatic acinar cells in response to physiological stimulation in vivo

The exocrine pancreas secretes fluid and digestive Enzymes in response to parasympathetic release of acetylcholine (ACh) via the vagus nerve and the gut hormone cholecystokinin (CCK). Both secretion of fluid and exocytosis of secretory granules containing Enzymes and zymogens are dependent on an increase in the cytosolic [CA²⁺ ] in acinar cells. It is thought that the specific spatiotemporal characteristics of the CA²⁺ signals are fundamental for appropriate secretion and that these properties are disrupted in disease states in the pancreas. While extensive research has been performed to characterize CA²⁺ signalling in acinar cells, this has exclusively been achieved in ex vivo preparations of exocrine cells, where it is difficult to mimic physiological conditions. Here we have developed a method to optically observe pancreatic acinar CA²⁺ signals in vivo using a genetically expressed CA²⁺ indicator and imaged with multi-photon microscopy in live Animals. In vivo, acinar cells exhibited baseline activity in fasted Animals, which was dependent on CCK1 receptors (CCK1Rs). Both stimulation of intrinsic nervous input and administration of systemic CCK induced oscillatory activity in a proportion of the cells, but the maximum frequencies were vastly different. Upon feeding, oscillatory activity was also observed, which was dependent on CCK1Rs. No evidence of a vago-vagal reflex mediating the effects of CCK was observed. Our in vivo method revealed the spatial and temporal profile of physiologically evoked CA²⁺ signals, which will provide new insights into future studies of the mechanisms underlying exocrine physiology and that are disrupted in pathological conditions. KEY POINTS: In the exocrine pancreas, the spatiotemporal properties of CA²⁺ signals are fundamentally important for the appropriate stimulation of secretion by the neurotransmitter acetylcholine and gut hormone cholecystokinin. These characteristics were previously defined in ex vivo studies. Here we report the spatiotemporal characteristics of CA²⁺ signals in vivo in response to physiological stimulation in a mouse engineered to express a CA²⁺ indicator in acinar cells. Specific CA²⁺ 'signatures' probably important for stimulating secretion are evoked in vivo in fasted Animals, by feeding, neural stimulation and cholecystokinin administration. The CA²⁺ signals are probably the result of the direct action of ACh and CCK on acinar cells and not indirectly through a vago-vagal reflex.

GCamp6F; acetylcholine; calcium signalling; cholecystokinin; exocrine secretion; intravital microscopy; multiphoton microscopy; pancreatic acinar cells.