期刊
SCIENCE CHINA-LIFE SCIENCES
卷 61, 期 11, 页码 1352-1368出版社
SCIENCE PRESS
DOI: 10.1007/s11427-018-9315-6
关键词
Ca2+ signaling; neurospheres; zebrafish; neural stem; progenitor cells; differentiation; IP3 receptors; ryanodine receptors; STIM and Orai
类别
资金
- ANR/RGC Joint Research Scheme Award [A-HKUST601/13]
- HK RGC General Research Fund [662113, 16101714, 16100115]
- HKITC [ITCPD/17-9]
The development of the CNS in vertebrate embryos involves the generation of different sub-types of neurons and glia in a complex but highly-ordered spatio-temporal manner. Zebrafish are commonly used for exploring the development, plasticity and regeneration of the CNS, and the recent development of reliable protocols for isolating and culturing neural stem/progenitor cells (NSCs/NPCs) from the brain of adult fish now enables the exploration of mechanisms underlying the induction/specification/differentiation of these cells. Here, we refined a protocol to generate proliferating and differentiating neurospheres from the entire brain of adult zebrafish. We demonstrated via RT-qPCR that some isoforms of ip3r, ryr and stim are upregulated/downregulated significantly in differentiating neurospheres, and via immunolabelling that 1,4,5-inositol trisphosphate receptor (IP3R) type-1 and ryanodine receptor (RyR) type-2 are differentially expressed in cells with neuron- or radial glial-like properties. Furthermore, ATP but not caffeine (IP3R and RyR agonists, respectively), induced the generation of Ca2+ transients in cells exhibiting neuron- or glial-like morphology. These results indicate the differential expression of components of the Ca2+-signaling toolkit in proliferating and differentiating cells. Thus, given the complexity of the intact vertebrate brain, neurospheres might be a useful system for exploring neurodegenerative disease diagnosis protocols and drug development using Ca2+ signaling as a read-out.
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