PUBLICATION
The glutathione degrading enzyme, Chac1, is required for calcium signaling in developing zebrafish: Redox as an upstream activator of calcium
- Authors
- Yadav, S., Chawla, B., Khursheed, M.A., Ramachandran, R., Bachhawat, A.K.
- ID
- ZDB-PUB-190614-4
- Date
- 2019
- Source
- The Biochemical journal 476(13): 1857-1873 (Journal)
- Registered Authors
- Keywords
- Chac1, Development, Glutathione, Glutathione redox potential, Zebrafish, calcium signalling
- MeSH Terms
-
- Animals
- Calcium/metabolism
- Calcium Signaling/physiology*
- Embryo, Nonmammalian/enzymology*
- Oxidation-Reduction
- Zebrafish/embryology*
- Zebrafish/genetics
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- gamma-Glutamylcyclotransferase/genetics
- gamma-Glutamylcyclotransferase/metabolism*
- PubMed
- 31189567 Full text @ Biochem. J.
Citation
Yadav, S., Chawla, B., Khursheed, M.A., Ramachandran, R., Bachhawat, A.K. (2019) The glutathione degrading enzyme, Chac1, is required for calcium signaling in developing zebrafish: Redox as an upstream activator of calcium. The Biochemical journal. 476(13):1857-1873.
Abstract
Calcium signaling is essential for embryonic development but the signals upstream of calcium are only partially understood. Here we investigate the role of the intracellular glutathione redox potential in calcium signaling using the Chac1 protein of zebrafish. A member of the Γ-glutamylcyclotransferase family of enzymes, the zebrafish Chac1 is a glutathione degrading enzyme that acts only on reduced glutathione. The zebrafish chac1 expression was seen early in development, and in the latter stages, in the developing muscles, brain and heart. The chac1 knockdown was embryonic lethal, and the developmental defects were seen primarily in the myotome, brain and heart where chac1 was maximally expressed. The phenotypes could be rescued by the WT Chac1 but not by the catalytically inactive Chac1 that was incapable of degrading glutathione. The ability of chac1 to alter the intracellular glutathione redox potential in the live animals was examined using Grx1-roGFP2. The chac1 morphants lacked the increased degree of cellular oxidation seen in the WT zebrafish. As calcium is also known to be critical for the developing myotomes, brain and heart, we further investigated if the chac1 knockdown phenotypes were a consequence of the lack of calcium signals. We observed using GCaMP6s, that calcium transients normally seen in the developing embryos were strongly attenuated in these knockdowns. The study thus identifies Chac1 and the consequent change in intracellular glutathione redox potential as important upstream activators of calcium signaling during development.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping