Neural Regeneration Research ›› 2021, Vol. 16 ›› Issue (11): 2257-2263.doi: 10.4103/1673-5374.310697

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Identification of potential candidate proteins for reprogramming spinal cord-derived astrocytes into neurons: a proteomic analysis

Wen-Hao Chen, Yu-Xiang Lin, Ling Lin, Bao-Quan Zhang, Shu-Xia Xu, Wei Wang#br#   

  1. 1Department of Pediatric Surgery, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 2Department of Breast Surgery, Affiliated Union Hospital, Fujian Medical University, Fuzhou, Fujian Province, China; 3Institutes of Biomedical Sciences of Shanghai Medical School, Fudan University, Shanghai, China; 4Department of Neonatology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 5Department of Pathology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, China; 6Department of Anatomy and Histoembryology, Fujian Medical University, Fuzhou, Fujian Province, China
  • Online:2021-11-15 Published:2021-04-13
  • Contact: Wen-Hao Chen, MD, 15606075620@163.com.
  • Supported by:
    The study was supported by the Natural Science Foundation of Fujian Province, China, No. 2015J05153; Research Talents Training Project of Fujian Provincial Health Department, China, No. 2018-ZQN-29; and Joint Funds for the Innovation of Science and Technology of Fujian Province, China, No. 2018Y9002 (all to WHC).

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Abstract: Our previous study has confirmed that astrocytes overexpressing neurogenic differentiation factor 1 (NEUROD1) in the spinal cord can be reprogrammed into neurons under in vivo conditions. However, whether they can also be reprogrammed into neurons under in vitro conditions remains unclear, and the mechanisms of programmed conversion from astrocytes to neurons have not yet been clarified. In the present study, we prepared reactive astrocytes from newborn rat spinal cord astrocytes using the scratch method and infected them with lentivirus carrying NEUROD1. The results showed that NEUROD1 overexpression reprogrammed the cultured reactive astrocytes into neurons in vitro with an efficiency of 13.4%. Using proteomic and bioinformatic analyses, 1952 proteins were identified, of which 92 were differentially expressed. Among these proteins, 11 were identified as candidate proteins in the process of reprogramming based on their biological functions and fold-changes in the bioinformatic analysis. Furthermore, western blot assay revealed that casein kinase II subunit alpha (CSNK2A2) and pinin (PNN) expression in NEUROD1-overexpressing reactive astrocytes was significantly increased, suggesting that NEUROD1 can directly reprogram spinal cord-derived reactive astrocytes into neurons in vitro, and that the NEUROD1-CSNK2A2-PNN pathway is involved in this process. This study was approved by the Animal Ethics Committee of Fujian Medical University, China (approval No. 2016-05) on April 18, 2016. 

Key words: astrocytes, glial cells, label-free proteomic analysis, mechanism, neurons, primary culture, reprogramming, spinal cord injury

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