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Delgadillo-Silva, L.F.* ; Akhtar, M.N.* ; Taşöz, E.* ; Ninov, N.

Simultaneous calcium imaging and glucose stimulation in living zebrafish to investigate in vivo β-cell function.

J. Vis. Exp. 2021:e62347 (2021)
DOI
The pancreatic β-cells sustain systemic glucose homeostasis by producing and secreting insulin according to the blood glucose levels. Defects in β-cell function are associated with hyperglycemia that can lead to diabetes. During the process of insulin secretion, β-cells experience an influx of Ca2+. Thus, imaging the glucose-stimulated Ca2+ influx using genetically encoded calcium indicators (GECIs) provides an avenue to studying β-cell function. Previously, studies showed that isolated zebrafish islets expressing GCaMP6s exhibit significant Ca2+ activity upon stimulation with defined glucose concentrations. However, it is paramount to study how β-cells respond to glucose not in isolation, but in their native environment, where they are systemically connected, vascularized, and densely innervated. To this end, the study leveraged the optical transparency of the zebrafish larvae at early stages of development to illuminate β-cell activity in vivo. Here, a detailed protocol for Ca2+ imaging and glucose stimulation to investigate β-cell function in vivo is presented. This technique allows to monitor the coordinated Ca2+ dynamics in β-cells with single-cell resolution. Additionally, this method can be applied to work with any injectable solution such as small molecules or peptides. Altogether, the protocol illustrates the potential of the zebrafish model to investigate islet coordination in vivo and to characterize how environmental and genetic components might affect β-cell function.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Ca2+ Dynamics; Islets
ISSN (print) / ISBN 1940-087X
e-ISSN 1940-087X
Quellenangaben Volume: 2021, Issue: 175, Pages: , Article Number: e62347 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)
Grants Immunological and Cellular Strategies in Metabolic Disease
International Research Training Group
German Research Foundation (DFG)
German Center for Diabetes Research (DZD)
Center for Regenerative Therapies Dresden at TU Dresden