PUBLICATION

Visualization, characterization and modulation of calcium signaling during the development of slow muscle cells in intact zebrafish embryos

Authors
Cheung, C.Y., Webb, S.E., Love, D.R., and Miller, A.L.
ID
ZDB-PUB-110523-16
Date
2011
Source
The International journal of developmental biology   55(2): 153-74 (Journal)
Registered Authors
Cheung, Yuk Kam Chris, Love, Donald R., Miller, Andrew L., Webb, Sarah E.
Keywords
Ca2+ imaging, IP3R, RyR, slow muscle cell, transgenic zebrafish
MeSH Terms
  • Actins/metabolism
  • Aequorin/biosynthesis
  • Aequorin/genetics
  • Animals
  • Animals, Genetically Modified
  • Apoproteins/biosynthesis
  • Apoproteins/genetics
  • Bungarotoxins/pharmacology
  • Calcium/metabolism
  • Calcium Signaling*
  • Colforsin/pharmacology
  • Inositol 1,4,5-Trisphosphate Receptors/metabolism
  • Muscle Development/drug effects
  • Muscle Development/physiology*
  • Muscle, Skeletal/drug effects
  • Muscle, Skeletal/embryology*
  • Muscle, Skeletal/metabolism
  • Myosins/metabolism
  • Nifedipine/pharmacology
  • Recombinant Proteins/biosynthesis
  • Recombinant Proteins/genetics
  • Ryanodine Receptor Calcium Release Channel/metabolism
  • Veratrum Alkaloids/pharmacology
  • Zebrafish/embryology*
  • Zebrafish/metabolism*
PubMed
21553383 Full text @ Int. J. Dev. Biol.
Abstract
Intact zebrafish embryos were used as an in vivo animal model to investigate the role of Ca(2+) signaling during the differentiation of slow muscle cells (SMCs) within forming skeletal muscle. Transgenic zebrafish were generated using an a-actin promoter that targeted apoaequorin expression specifically to muscle cells. Two distinct Ca(2+) signaling periods (CSPs) were visualized in the developing SMCs: between ~17.5-19.5 hours post-fertilization (hpf) and after ~23 hpf, separated by a ~3.5 h Ca(2+) signaling quiet period. Further spatial characterization of these Ca2+ signals using confocal fluorescent microscopy and calcium green-1 dextran as a reporter, indicated that the earlier CSP displayed distinct nuclear and cytoplasmic components, whereas the later CSP was predominantly cytoplasmic. Both CSPs consisted of a series of oscillating Ca(2+) waves generated at distinct frequencies, while the earlier CSP also displayed a slow rise then fall in the Ca(2+) baseline-level. Imaging of cyclopamine- and forskolin-treated wild-type, or smo(-/-) mutant embryos, where SMCs do not form, confirmed the specific cell population generating the signals. Treating embryos with antagonists indicated that both IP(3)Rs and RyRs are responsible for generating the temporal characteristics of the Ca(2+) signaling signature, and that the latter plays a necessary role in SMC differentiation and subsequent myotome patterning. Together, these data support and extend the proposition that specific spatiotemporal patterns of spontaneous Ca(2+) signals might be used for different as well as combinatorial regulation of both nuclear and cytosolic signal transduction cascades, resulting in myofibrillogenesis in SMCs as well as myotome patterning.
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