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Janjuha, S. ; Pal Singh, S.* ; Ninov, N.

Analysis of beta-cell function using single-cell resolution calcium imaging in zebrafish islets.

J. Vis. Exp.:e57851 (2018)
Publ. Version/Full Text Research data DOI
Open Access Gold (Paid Option)
Creative Commons Lizenzvertrag
Pancreatic beta-cells respond to increasing blood glucose concentrations by secreting the hormone insulin. The dysfunction of beta-cells leads to hyperglycemia and severe, life-threatening consequences. Understanding how the beta-cells operate under physiological conditions and what genetic and environmental factors might cause their dysfunction could lead to better treatment options for diabetic patients. The ability to measure calcium levels in beta-cells serves as an important indicator of beta-cell function, as the influx of calcium ions triggers insulin release. Here we describe a protocol for monitoring the glucose-stimulated calcium influx in zebrafish beta-cells by using GCaMP6s, a genetically encoded sensor of calcium. The method allows monitoring the intracellular calcium dynamics with single-cell resolution in ex vivo mounted islets. The glucose-responsiveness of beta-cells within the same islet can be captured simultaneously under different glucose concentrations, which suggests the presence of functional heterogeneity among zebrafish beta-cells. Furthermore, the technique provides high temporal and spatial resolution, which reveals the oscillatory nature of the calcium influx upon glucose stimulation. Our approach opens the doors to use the zebrafish as a model to investigate the contribution of genetic and environmental factors to beta-cell function and dysfunction.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Biology ; Issue 137 ; Beta-cells ; Calcium ; Gcamp ; Live-imaging ; Function ; Diabetes ; Zebrafish; Insulin-release; Pancreatic-islets; Glucose; Nitroreductase; Oscillations; Langerhans; Ablation; Ca2+
ISSN (print) / ISBN 1940-087X
e-ISSN 1940-087X
Quellenangaben Volume: , Issue: 137, Pages: , Article Number: e57851 Supplement: ,
Publisher JoVE
Publishing Place 1 Alewife Center, Ste 200, Cambridge, Ma 02140 Usa
Reviewing status Peer reviewed
Institute(s) Institute for Pancreatic Beta Cell Research (IPI)