In vivo monitoring of intracellular Ca2+ dynamics in the pancreatic β-cells of zebrafish embryos.
Reka LorinczChristopher H EmfingerAndrea WalcherMichael GiolaiClaudia KrautgasserMaria S RemediColin G NicholsDirk MeyerPublished in: Islets (2018)
Assessing the response of pancreatic islet cells to glucose stimulation is important for understanding β-cell function. Zebrafish are a promising model for studies of metabolism in general, including stimulus-secretion coupling in the pancreas. We used transgenic zebrafish embryos expressing a genetically-encoded Ca2+ sensor in pancreatic β-cells to monitor a key step in glucose induced insulin secretion; the elevations of intracellular [Ca2+]i. In vivo and ex vivo analyses of [Ca2+]i demonstrate that β-cell responsiveness to glucose is well established in late embryogenesis and that embryonic β-cells also respond to free fatty acid and amino acid challenges. In vivo imaging of whole embryos further shows that indirect glucose administration, for example by yolk injection, results in a slow and asynchronous induction of β-cell [Ca2+]i responses, while intravenous glucose injections cause immediate and islet-wide synchronized [Ca2+]i fluctuations. Finally, we demonstrate that embryos with disrupted mutation of the CaV1.2 channel gene cacna1c are hyperglycemic and that this phenotype is associated with glucose-independent [Ca2+]i fluctuation in β-cells. The data reveal a novel central role of cacna1c in β-cell specific stimulus-secretion coupling in zebrafish and demonstrate that the novel approach we propose - to monitor the [Ca2+]i dynamics in embryonic β-cells in vivo - will help to expand the understanding of β-cell physiological functions in healthy and diseased states.
Keyphrases
- induced apoptosis
- cell cycle arrest
- blood glucose
- cell therapy
- cell death
- protein kinase
- stem cells
- cell proliferation
- mesenchymal stem cells
- adipose tissue
- high resolution
- low dose
- skeletal muscle
- transcription factor
- photodynamic therapy
- endothelial cells
- artificial intelligence
- fluorescence imaging
- weight loss
- ultrasound guided
- electronic health record
- genome wide identification
- case control