Analysis of Beta-cell Function Using Single-cell Resolution Calcium Imaging in Zebrafish Islets
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
Abstract
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.
Details
Original language | English |
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Article number | e57851 |
Number of pages | 6 |
Journal | JoVE |
Volume | 137 |
Publication status | Published - Jul 2018 |
Peer-reviewed | Yes |
External IDs
PubMed | 30035763 |
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Scopus | 85049828950 |
Keywords
Sustainable Development Goals
Keywords
- Beta-cells, Biology, GCaMP, Issue 137, Calcium, Diabetes, Function, Live-imaging, Zebrafish