Generation of GCaMP6s-Expressing Zebrafish to Monitor Spatiotemporal Dynamics of Calcium Signaling Elicited by Heat Stress

Li, F., Long, Y., Xie, J., Ren, J., Zhou, T., Song, G., Li, Q., Cui, Z.
International Journal of Molecular Sciences   22(11): (Journal)
Registered Authors
Cui, Zongbin
GCaMP6s, calcium signaling, heat shock response, heat stress, transgenic zebrafish
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Calcium/metabolism*
  • Calcium Signaling*/drug effects
  • Calcium Signaling*/genetics
  • Calcium Signaling*/physiology
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2/antagonists & inhibitors
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism
  • Calmodulin/genetics
  • Calmodulin/metabolism*
  • Capsaicin/analogs & derivatives
  • Capsaicin/pharmacology
  • Green Fluorescent Proteins/genetics
  • Green Fluorescent Proteins/metabolism*
  • Heat Shock Transcription Factors/antagonists & inhibitors
  • Heat Shock Transcription Factors/metabolism
  • Heat-Shock Proteins/genetics
  • Heat-Shock Proteins/metabolism
  • Heat-Shock Response/genetics*
  • Heat-Shock Response/physiology
  • In Situ Hybridization
  • Lanthanum/pharmacology
  • Microscopy, Fluorescence
  • Spatio-Temporal Analysis
  • Zebrafish/genetics*
  • Zebrafish/growth & development
  • Zebrafish/metabolism
  • Zebrafish Proteins/antagonists & inhibitors
  • Zebrafish Proteins/metabolism
34074030 Full text @ Int. J. Mol. Sci.
The ability of organisms to quickly sense and transduce signals of environmental stresses is critical for their survival. Ca2+ is a versatile intracellular messenger involved in sensing a wide variety of stresses and regulating the subsequent cellular responses. So far, our understanding for calcium signaling was mostly obtained from ex vivo tissues and cultured cell lines, and the in vivo spatiotemporal dynamics of stress-triggered calcium signaling in a vertebrate remains to be characterized. Here, we describe the generation and characterization of a transgenic zebrafish line with ubiquitous expression of GCaMP6s, a genetically encoded calcium indicator (GECI). We developed a method to investigate the spatiotemporal patterns of Ca2+ events induced by heat stress. Exposure to heat stress elicited immediate and transient calcium signaling in developing zebrafish. Cells extensively distributed in the integument of the head and body trunk were the first batch of responders and different cell populations demonstrated distinct response patterns upon heat stress. Activity of the heat stress-induced calcium signaling peaked at 30 s and swiftly decreased to near the basal level at 120 s after the beginning of exposure. Inhibition of the heat-induced calcium signaling by LaCl3 and capsazepine and treatment with the inhibitors for CaMKII (Ca²2/calmodulin-dependent protein kinase II) and HSF1 (Heat shock factor 1) all significantly depressed the enhanced heat shock response (HSR). Together, we delineated the spatiotemporal dynamics of heat-induced calcium signaling and confirmed functions of the Ca2+-CaMKII-HSF1 pathway in regulating the HSR in zebrafish.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes