PUBLICATION
In vivo monitoring of intracellular Ca2+ dynamics in the pancreatic β-cells of zebrafish embryos
- Authors
- Lorincz, R., Emfinger, C.H., Walcher, A., Giolai, M., Krautgasser, C., Remedi, M.S., Nichols, C.G., Meyer, D.
- ID
- ZDB-PUB-181207-28
- Date
- 2018
- Source
- Islets 10(6): 221-238 (Journal)
- Registered Authors
- Krautgasser, Claudia, Lorincz, Réka, Meyer, Dirk
- Keywords
- Cav1.2 channel, GCaMP6s, cacna1c, early zebrafish development, glucose-sensing of beta cells, in vivo imaging
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Calcium/metabolism*
- Calcium Channels, L-Type/metabolism*
- Embryo, Nonmammalian/metabolism*
- Insulin-Secreting Cells/metabolism*
- Zebrafish
- PubMed
- 30521410 Full text @ Islets
Citation
Lorincz, R., Emfinger, C.H., Walcher, A., Giolai, M., Krautgasser, C., Remedi, M.S., Nichols, C.G., Meyer, D. (2018) In vivo monitoring of intracellular Ca2+ dynamics in the pancreatic β-cells of zebrafish embryos. Islets. 10(6):221-238.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping