Neural Regeneration Research ›› 2023, Vol. 18 ›› Issue (3): 577-581.doi: 10.4103/1673-5374.350208

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DUSP2 deletion with CRISPR/Cas9 promotes Mauthner cell axonal regeneration at the early stage of zebrafish

Guo-Jian Shao1, #, Xin-Liang Wang1, #, Mei-Li Wei2, Da-Long Ren2, *, Bing Hu1, *   

  1. 1School of Life Sciences, Division of Biomedical Sciences, University of Science and Technology of China, Hefei, Anhui Province, China; 2Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei, Anhui Province, China
  • Online:2023-03-15 Published:2022-08-26
  • Contact: Da-Long Ren, PhD, rendl@ustc.edu.cn; Bing Hu, PhD, bhu@ustc.edu.cn.
  • Supported by:
    This work was granted by the National Natural Science Foundation of China, No. 82071357 and Ministry of Science and Technology of China, No. 2019YFA0405600 (both to BH).

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Abstract: Axon regeneration of central neurons is a complex process that is tightly regulated by multiple extrinsic and intrinsic factors. The expression levels of distinct genes are changed after central neural system (CNS) injury and affect axon regeneration. A previous study identified dusp2 as an upregulated gene in zebrafish with spinal cord injury. Here, we found that dual specificity phosphatase 2 (DUSP2) is a negative regulator of axon regeneration of the Mauthner cell (M-cell). DUSP2 is a phosphatase that mediates the dephosphorylation of JNK. In this study, we knocked out dusp2 by CRISPR/Cas9 and found that M-cell axons of dusp2–/– zebrafish had a better regeneration at the early stage after birth (within 8 days after birth), while those of dusp2+/– zebrafish did not. Overexpression of DUSP2 in Tg (Tol 056)  zebrafish by single-cell electroporation retarded the regeneration of M-cell axons. Western blotting results showed that DUSP2 knockout slightly increased the levels of phosphorylated JNK. These findings suggest that knocking out DUSP2 promoted the regeneration of zebrafish M-cell axons, possibly through enhancing JNK phosphorylation. 

Key words: axon regeneration, central nervous system, CRISPR/Cas9, DUSP2, JNK, Mauthner cell, single-cell electroporation, spinal cord injury, two-photon axotomy, zebrafish