中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (7): 1258-1265.doi: 10.4103/1673-5374.301035

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

脊髓损伤后的皮质转录组分析揭示坍塌反应调节蛋白2敲入小鼠的中枢神经系统再生机制

  

  • 出版日期:2021-07-15 发布日期:2021-01-07
  • 基金资助:
    日本教育，文化，体育，科学和技术部（26430043）优先领域科学研究补助金支持

Cortical transcriptome analysis after spinal cord injury reveals the regenerative mechanism of central nervous system in CRMP2 knock-in mice

Ayaka Sugeno1, 3, Wenhui Piao1, Miki Yamazaki2, 3, Kiyofumi Takahashi4, Koji Arikawa4, Hiroko Matsunaga4, Masahito Hosokawa2, 4, 5, Daisuke Tominaga3, 6, Yoshio Goshima7, Haruko Takeyama2, 3, 4, 5, Toshio Ohshima1, 5, *   

  1. 1Laboratory for Molecular Brain Science, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan; 2Biomolecular Engineering Laboratory, Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan; 3Computational Bio Big-Data Open Innovation Laboratory (CBBD-OIL), National Institute of Advanced Industrial Science and Technology (AIST), Tokyo, Japan; 4Research Organization for Nano and Life Innovation, Waseda University, Tokyo, Japan; 5Institute for Advanced Research of Biosystem Dynamics, Waseda Research Institute for Science and Engineering, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan; 6Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan; 7Department of Molecular Pharmacology and Neurobiology, Graduate School of Medicine, Yokohama City University, Yokohama, Japan
  • Online:2021-07-15 Published:2021-01-07
  • Contact: Toshio Ohshima, MD, PhD, ohshima@waseda.jp.
  • Supported by:
    This work was supported by Grants-in-Aid for Scientific Research on Priority Areas from The Ministry of Education, Culture, Sports, Science and Technology (No. 26430043; to TO).

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摘要：

最近的研究表明，Ser522处的突变导致抑制坍塌反应调节蛋白2（CRMP2）磷酸化，并诱导轴延长和脊髓损伤后丧失的感觉运动功能的部分恢复。实验旨在通过使用显微解剖穿孔系统在小鼠感觉运动皮质中进行转录组分析，揭示CRMP2基因敲入（CRMP2KI）小鼠模型中轴突神经元再生的细胞内机制。实验发现脊髓损伤后轴索切除或邻近神经元的病理生理结构变化，发现在CRMP2基因敲入小鼠中大脑感觉运动皮质神经细胞胞体萎缩和树突棘减少受到抑制。转录组的进一步分析鉴定了脊髓损伤后4种野生型小鼠中显著上调的血红蛋白基因Hba-a1，Hba-a2，Hbb-bs和Hbb-bt，其同时伴随涉及氧化磷酸化和核糖体途径基因的上调。在CRMP2基因敲入小鼠中观察到通道活性基因表达的大量上调和调节囊泡、突触功能、神经胶质细胞分化基因表达的下调。同时CRMP2基因敲入小鼠的转录组分析发现能量代谢和神经元功能途径的差异调节。以上结果表明，通过提高神经元微管稳定性，调节中枢神经系统的整个代谢系统、神经胶质细胞和突触功能，CRMP2基因敲入小鼠脊髓损伤病理生理改变得以改善。

https://orcid.org/0000-0003-4931-7087 (Toshio Ohshima)

Abstract: Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2 (CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function after spinal cord injury (SCI). We aimed to reveal the intracellular mechanism in axotomized neurons in the CRMP2 knock-in (CRMP2KI) mouse model by performing transcriptome analysis in mouse sensorimotor cortex using micro-dissection punching system. Prior to that, we analyzed the structural pathophysiology in axotomized or neighboring neurons after SCI and found that somatic atrophy and dendritic spine reduction in sensorimotor cortex were suppressed in CRMP2KI mice. Further analysis of the transcriptome has aided in the identification of four hemoglobin genes Hba-a1, Hba-a2, Hbb-bs, and Hbb-bt that are significantly upregulated in wild-type mice with concomitant upregulation of genes involved in the oxidative phosphorylation and ribosomal pathways after SCI. However, we observed substantial upregulation in channel activity genes and downregulation of genes regulating vesicles, synaptic function, glial cell differentiation in CRMP2KI mice. Moreover, the transcriptome profile of CRMP2KI mice has been discussed wherein energy metabolism and neuronal pathways were found to be differentially regulated. Our results showed that CRMP2KI mice displayed improved SCI pathophysiology not only via microtubule stabilization in neurons, but also possibly via the whole metabolic system in the central nervous system, response changes in glial cells, and synapses. Taken together, we reveal new insights on SCI pathophysiology and the regenerative mechanism of central nervous system by the inhibition of CRMP2 phosphorylation at Ser522. All these experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee at Waseda University, Japan (2017-A027 approved on March 21, 2017; 2018-A003 approved on March 25, 2018; 2019-A026 approved on March 25, 2019).

Key words: CNS regeneration, cortex, CRMP2, hemoglobin, metabolic pathway, spinal cord injury, spine, transcriptome