PUBLICATION

Transgenic zebrafish for ratiometric imaging of cytosolic and mitochondrial Ca2+ response in teleost embryo

Authors
Mizuno, H., Sassa, T., Higashijima, S.I., Okamoto, H., and Miyawaki, A.
ID
ZDB-PUB-130816-46
Date
2013
Source
Cell Calcium   54(3): 236-45 (Journal)
Registered Authors
Higashijima, Shin-ichi, Miyawaki, Atsushi, Okamoto, Hitoshi
Keywords
calcium, zebrafish, embryo, fertilization, mitochondrial calcium, FRET
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Calcium/metabolism*
  • Calcium Signaling
  • Calcium-Binding Proteins/metabolism
  • Cytosol/metabolism*
  • Embryo, Nonmammalian/metabolism*
  • HSC70 Heat-Shock Proteins/metabolism
  • Microscopy, Confocal
  • Mitochondria/metabolism*
  • Muscle Cells/physiology
  • Muscle Contraction/physiology
  • Zebrafish/growth & development
  • Zebrafish/metabolism*
  • Zebrafish Proteins/metabolism
PubMed
23906585 Full text @ Cell Calcium
Abstract

Intracellular Ca2+ imaging has widely been used to visualize intracellular signals, but the application in an intact animal is still limited due to difficulty of the indicator loading. In addition, the motion of the living animal produces artifacts. To investigate Ca2+ signaling at early embryonic stage, we established transgenic zebrafish line expressing a genetically encoded Ca2+ indicator, cameleon YC2.60, driven by a constitutively active promoter, hspa8. Although the embryo dynamically changes its morphology, the motion artifact could be canceled out by taking the advantage of YC2.60 as a ratiometric indicator. The transgenic zebrafish was used to visualize the propagation of cytosolic Ca2+ during the early embryonic stage upon fertilization and along cleavage furrow, and the rise in Ca2+ in the myocytes contracting spontaneously in the embryo. We also established a transgenic zebrafish line expressing YC2.60 targeted to the mitochondria. The rise in mitochondrial Ca2+ was rather sustained (H2 min), which is consistent with the requirement of ATP refilling since the mitochondrial Ca2+ upregulates rate-limiting enzymes of Krebs cycle. This is in contrast with the transient rise in the cytosol Ca2+ that directly evokes the muscle contraction. These transgenic zebrafish lines are expected to serve as useful tools further Ca2+ imaging in vivo.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping