中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (2): 270-276.doi: 10.4103/1673-5374.265561

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

胶质细胞源性神经营养因子剪接体在小鼠脑中的差异表达

  

  • 出版日期:2020-02-15 发布日期:2020-05-25
  • 基金资助:
    国家自然科学基金资助项目（81772688）; 中国江苏省博士后科学基金（1202119C）

Differential expression of glial cell line-derived neurotrophic factor splice variants in the mouse brain

Xiao-He Gu1, Heng Li1, Lin Zhang1, Tao He1, Xiang Chai1, He Wei2, Dian-Shuai Gao1   

  1. 1 Department of Anatomy and Neurobiology, Xuzhou Medical University, Xuzhou, Jiangsu Province, China
    2 Department of Neurosurgery, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China
  • Online:2020-02-15 Published:2020-05-25
  • Contact: Dian-Shuai Gao, MD,gds@xzhmu.edu.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, No. 81772688 (to DSG); the Postdoctoral Science Foundation of Jiangsu Province of China, No. 1202119C (to HL).

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摘要：


胶质细胞源性神经营养因子（GDNF）在神经元存活和功能中起重要作用，有α-pro-GDNF和β-pro-GDNF两种剪切体，这两种剪切体对多巴胺神经元具有重要的保护作用。但是，目前仍然缺乏GDNF剪接体在富含多巴胺能神经元的脑区中表达的研究。(1)为了确定正常小鼠脑中α-和β-pro-GDNF的mRNA表达，实验成功设计并制备了GDNF剪接体的特异性引物，随后进行实时荧光定量PCR检测发现，小鼠脑组织β-pro-GDNF的mRNA表达显著强于α-pro-GDNF的mRNA表达。为了评估小鼠脑组织中α-和β-pro-GDNF的蛋白表达，进行了免疫印迹检测。结果显示，β-pro-GDNF的蛋白表达显著低于α-pro-GDNF的蛋白表达，与mRNA表达结果相反；(2)为了探索GDNF剪接体转录和翻译水平反向表达的可能原因，用实时荧光定量PCR分析了GDNF剪接体与分选蛋白相关受体A（SorLA）mRNA表达的关系。发现小鼠脑中SorLA mRNA与β-pro-GDNF mRNA表达呈正相关，但与α-pro-GDNF mRNA表达无关；(3)上述数据显示，α-和β-pro-GDNF剪接体在小鼠脑中呈差异表达。SorLA作为一种分选蛋白，可能导致GDNF剪接异构体mRNA与蛋白的反向表达。实验已于2016年7月14日经中国徐州医科大学动物伦理委员会批准。

orcid: 0000-0001-8567-0238 (Dian-Shuai Gao)

关键词:

胶质细胞源性神经营养因子, 剪切体, 胶质细胞源性神经营养因子前体α, 胶质细胞源性神经营养因子前体β, 分选蛋白相关受体A, 小鼠脑, 脑区, 多巴胺能神经元, Δ78位点, 前体蛋白, 神经再生

Abstract: Glial cell line-derived neurotrophic factor (GDNF) plays a critical role in neuronal survival and function. GDNF has two major splice variants in the brain, α-pro-GDNF and β-pro-GDNF, and both isoforms have strong neuroprotective effects on dopamine neurons. However, the expression of the GDNF splice variants in dopaminergic neurons in the brain remains unclear. Therefore, in this study, we investigated the mRNA and protein expression of α- and β-pro-GDNF in the mouse brain by real-time quantitative polymerase chain reaction, using splice variant-specific primers, and western blot analysis. At the mRNA level, β-pro-GDNF expression was significantly greater than that of α-pro-GDNF in the mouse brain. In contrast, at the protein level, α-pro-GDNF expression was markedly greater than that of β-pro-GDNF. To clarify the mechanism underlying this inverse relationship in mRNA and protein expression levels of the GDNF splice variants, we analyzed the expression of sorting protein-related receptor with A-type repeats (SorLA) by real-time quantitative polymerase chain reaction. At the mRNA level, SorLA was positively associated with β-pro-GDNF expression, but not with α-pro-GDNF expression. This suggests that the differential expression of α- and β-pro-GDNF in the mouse brain is related to SorLA expression. As a sorting protein, SorLA could contribute to the inverse relationship among the mRNA and protein levels of the GDNF isoforms. This study was approved by the Animal Ethics Committee of Xuzhou Medical University, China on July 14, 2016.

Key words: Δ78 locus, brain region, dopaminergic neurons, glial cell line-derived neurotrophic factor, mouse brain, precursor protein, α-pro-GDNF, β-pro-GDNF, sorting protein-related receptor with A-type repeats, splice variants