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Sensors 2015, 15(11), 27393-27419; doi:10.3390/s151127393

Membrane Potential and Calcium Dynamics in Beta Cells from Mouse Pancreas Tissue Slices: Theory, Experimentation, and Analysis

1
Institute of Physiology, Faculty of Medicine, University of Maribor, SI-2000 Maribor, Slovenia
2
Faculty of Electrical Engineering and Computer Science, University of Maribor, SI-2000 Maribor, Slovenia
3
Center for Open Innovation and Research, Core@UM, University of Maribor, SI-2000 Maribor, Slovenia
4
Department of Physics, Faculty of Natural Sciences and Mathematics, University of Maribor, SI-2000 Maribor, Slovenia
5
Center for Physiology and Pharmacology, Medical University of Vienna, A-1090 Vienna, Austria
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Frances S. Ligler
Received: 28 August 2015 / Revised: 11 October 2015 / Accepted: 14 October 2015 / Published: 28 October 2015
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Abstract

Beta cells in the pancreatic islets of Langerhans are precise biological sensors for glucose and play a central role in balancing the organism between catabolic and anabolic needs. A hallmark of the beta cell response to glucose are oscillatory changes of membrane potential that are tightly coupled with oscillatory changes in intracellular calcium concentration which, in turn, elicit oscillations of insulin secretion. Both membrane potential and calcium changes spread from one beta cell to the other in a wave-like manner. In order to assess the properties of the abovementioned responses to physiological and pathological stimuli, the main challenge remains how to effectively measure membrane potential and calcium changes at the same time with high spatial and temporal resolution, and also in as many cells as possible. To date, the most wide-spread approach has employed the electrophysiological patch-clamp method to monitor membrane potential changes. Inherently, this technique has many advantages, such as a direct contact with the cell and a high temporal resolution. However, it allows one to assess information from a single cell only. In some instances, this technique has been used in conjunction with CCD camera-based imaging, offering the opportunity to simultaneously monitor membrane potential and calcium changes, but not in the same cells and not with a reliable cellular or subcellular spatial resolution. Recently, a novel family of highly-sensitive membrane potential reporter dyes in combination with high temporal and spatial confocal calcium imaging allows for simultaneously detecting membrane potential and calcium changes in many cells at a time. Since the signals yielded from both types of reporter dyes are inherently noisy, we have developed complex methods of data denoising that permit for visualization and pixel-wise analysis of signals. Combining the experimental approach of high-resolution imaging with the advanced analysis of noisy data enables novel physiological insights and reassessment of current concepts in unprecedented detail. View Full-Text
Keywords: calcium sensors; membrane potential sensors; calcium imaging; membrane potential imaging; beta cell; pancreas; denoising; patch-clamp calcium sensors; membrane potential sensors; calcium imaging; membrane potential imaging; beta cell; pancreas; denoising; patch-clamp
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Dolenšek, J.; Špelič, D.; Klemen, M.S.; Žalik, B.; Gosak, M.; Rupnik, M.S.; Stožer, A. Membrane Potential and Calcium Dynamics in Beta Cells from Mouse Pancreas Tissue Slices: Theory, Experimentation, and Analysis. Sensors 2015, 15, 27393-27419.

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