Although the regulation of stress granules has become an intensely studied topic, current investigations of stress granule assembly, disassembly and dynamics are mainly performed in cultured cells. Here we report the establishment of a stress granule reporter to facilitate the real-time study of stress granules in vivo. Using CRISPR/Cas9, we fused a green fluorescence protein (GFP) to the endogenous G3BP1 in zebrafish. The GFP-G3BP1 reporter faithfully and robustly responded to heat stress in zebrafish embryos and larvae. The induction of stress granules varied by brain regions under the same stress condition, with the midbrain cells showing the highest efficiency and dynamics. Furthermore, preconditioning using lower heat stress significantly limited stress granule formation during subsequent heat stress. More interestingly, the stress granule formation was much more robust in zebrafish embryos than in larvae and coincided with significant elevated phosphorylated eIF2α and enhanced heat resilience. Therefore, these findings have generated new insights into stress response in zebrafish during early development and demonstrated that the GFP-G3BP1 knockin zebrafish could be a valuable tool for the investigation of stress granule biology.
Heat resilience in embryonic zebrafish revealed by an in vivo stress granule reporter
Present address: Department of Neuroscience, Baylor College of Medicine, Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, 1250 Moursund St, Houston, TX 77030, USA
Currently Viewing Accepted Manuscript - Newer Version Available
Ruiqi Wang, Hefei Zhang, Jiulin Du, Jin Xu; Heat resilience in embryonic zebrafish revealed by an in vivo stress granule reporter. J Cell Sci 2019; jcs.234807. doi: https://doi.org/10.1242/jcs.234807
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