Journal
FASEB JOURNAL
Volume 34, Issue 1, Pages 945-959Publisher
FEDERATION AMER SOC EXP BIOL
DOI: 10.1096/fj.201901302RR
Keywords
calcium dynamics; diabetes mellitus; fluorescence microscopy; in vivo imaging; pancreatic islets
Categories
Funding
- Svenska Forskningsradet Formas (Swedish Research Council Formas)
- Erling-Persson Foundation
- Stichting af Jochnick Foundation (Jochnick Foundation)
- Novo Nordisk Foundation Center for Basic Metabolic Research (NovoNordisk Foundation Center for Basic Metabolic Research)
- Swedish Diabetes Association
- Scandia Insurance Company Ltd
- Berth von Kantzow's Foundation
- Strategic Research Program in Diabetes at Karolinska Institutet
- BetaImage ERC-2013-AdG [338936]
- Swedish Foundation for Strategic Research
- Knut och Alice Wallenbergs Stiftelse (Knut and Alice Wallenberg Foundation)
- EYELETS ERC-2018-AdG [834860]
- BETASCREEN ERC-2017-PoC [727306]
- Diabetes Research and Wellness Foundation
- European Research Council (ERC) [834860, 338936, 727306] Funding Source: European Research Council (ERC)
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The dynamics of cytoplasmic free Ca2+ concentration ([Ca2+](i)) in pancreatic beta cells is central to our understanding of beta-cell physiology and pathology. In this context, there are numerous in vitro studies available but existing in vivo data are scarce. We now critically evaluate the anterior chamber of the eye as an in vivo, non-invasive, imaging site for measuring [Ca2+](i) dynamics longitudinally in three dimensions and at single-cell resolution. By applying a fluorescently labeled glucose analogue 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose in vivo, we followed how glucose almost simultaneously distributes to all cells within the islet volume, resulting in [Ca2+](i) changes. We found that almost all beta cells in healthy mice responded to a glucose challenge, while in hyperinsulinemic, hyperglycemic mice about 80% of the beta cells could not be further stimulated from fasting basal conditions. This finding indicates that our imaging modality can resolve functional heterogeneity within the beta-cell population in terms of glucose responsiveness. Importantly, we demonstrate that glucose homeostasis is markedly affected using isoflurane compared to hypnorm/midazolam anesthetics, which has major implications for [Ca2+](i) measurements. In summary, this setup offers a powerful tool to further investigate in vivo pancreatic beta-cell [Ca2+](i) response patterns at single-cell resolution in health and disease.
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