PUBLICATION
Reconstitution of holo-aequorin with apoaequorin mRNA and coelenterazine in zebrafish embryos
- Authors
- Chan, C.M., Miller, A.L., and Webb, S.E.
- ID
- ZDB-PUB-130605-10
- Date
- 2013
- Source
- Cold Spring Harbor protocols 2013(5): pdb.prot072975 (Journal)
- Registered Authors
- Miller, Andrew L., Webb, Sarah E.
- Keywords
- none
- MeSH Terms
-
- Aequorin/genetics
- Aequorin/metabolism*
- Animals
- Apoproteins/genetics
- Apoproteins/metabolism*
- Embryo, Nonmammalian
- Gene Expression*
- Imidazoles/metabolism*
- Microinjections/methods
- Pyrazines/metabolism*
- RNA, Messenger/genetics
- RNA, Messenger/isolation & purification
- RNA, Messenger/metabolism*
- Recombinant Proteins/genetics
- Recombinant Proteins/metabolism
- Zebrafish/embryology*
- PubMed
- 23637361 Full text @ Cold Spring Harb. Protoc.
Citation
Chan, C.M., Miller, A.L., and Webb, S.E. (2013) Reconstitution of holo-aequorin with apoaequorin mRNA and coelenterazine in zebrafish embryos. Cold Spring Harbor protocols. 2013(5):pdb.prot072975.
Abstract
When holo-aequorin is injected into zebrafish embryos at the one-cell stage, it is normally depleted by <24 h post-fertilization (hpf). In order to acquire Ca2+ signaling information from embryos older than 24 hpf, we have developed a protocol to express apoaequorin transiently in embryos, after which we reconstitute active holo-aequorin in vivo by introducing the cofactor coelenterazine into the developing embryo. This protocol describes the preparation of apoaequorin mRNA, followed by microinjection into embryos and incubation with coelenterazine to reconstitute holo-aequorin.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping