中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (3): 486-492.doi: 10.4103/1673-5374.202920

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

腺病毒介导模型小鼠显性中间C型腓骨肌萎缩症相关突变基因的解剖学分布缺陷

  

  • 收稿日期:2017-03-07 出版日期:2017-03-15 发布日期:2017-03-15
  • 基金资助:

    韩国国家研究基金

Anatomical distributional defects in mutant genes associated with dominant intermediate Charcot-Marie-Tooth disease type C in an adenovirus-mediated mouse model

SeoJin Lee1, Sandesh Panthi1, Hyun Woo Jo1, Jaeyoung Cho2, Min-Sik Kim3, Na Young Jeong4, In Ok Song5, Junyang Jung1, 2, 6, Youngbuhm Huh1, 2, 6   

  1. 1 Department of Biomedical Science, Graduate School, Kyung Hee University, Dongdaemun-gu, Seoul, Korea; 2 Department of Medicine, Graduate School, Kyung Hee University, Dongdaemun-gu, Seoul, Korea; 3 Department of Applied Chemistry, College of Applied Science, Kyung Hee University, Yongin-si, Gyeonggi-do, Korea; 4 Department of Anatomy and Cell Biology, College of Medicine, Dong-A University, Seo-gu, Busan, Korea; 5 Department of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Cheil General Hospital, Dankook University College of Medicine, Jung-gu, Seoul, Korea; 6 Department of Anatomy and Neurobiology, College of Medicine, Kyung Hee University, Dongdaemun-gu, Seoul, Korea
  • Received:2017-03-07 Online:2017-03-15 Published:2017-03-15
  • Contact: Youngbuhm Huh, M.D., Ph.D., jjung@khu.ac.kr or ybhuh@khu.ac.kr.
  • Supported by:

    This work was supported by the National Research Foundation (NRF) of Korea grant funded by Korean Government (MEST) (No. 2011-0030072).

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摘要：

显性中间C型腓骨肌萎缩症是一种表现为轴突和脱髓鞘的显性遗传神经病变，与酪氨酰-tRNA合成酶（YARS）杂合错义基因E196K和G41R突变有关。实验应用腺病毒载体系统和体内技术在YARS相关神经病变模型小鼠的脊髓、周围轴突和背根神经节内表达携带flag-tag的YARS融合蛋白，在携带YARS基因腺病毒载体转染后，比较各检测部位的野生型YARS和E196K突变蛋白的表达。（1）E196K 突变型小鼠背根神经节神经元中Flag/绿色荧光蛋白阳性表达的比例与野生型小鼠相近；（2）转染与未转染YARS基因腺病毒载体的小鼠脊髓腹角均有GFP阳性表达；（3）从而证实腺病毒介导的YARS突变基因在小鼠体内分布存在解剖学差异，提示YARS相关神经病变小鼠模型可用于显性中间C型腓骨肌萎缩症的研究。

ORCID:0000-0003-3946-5406(Junyang Jung); 0000-0002-7687-0374(Youngbuhm Huh)

关键词: 酪氨酰-tRNA合成酶, YARS基因突变, YARS相关神经病变, 腓骨肌萎缩症, 腺病毒介导的小鼠模型

Abstract:

Dominant intermediate Charcot-Marie-Tooth disease type C (DI-CMTC) is a dominantly inherited neuropathy that has been classified primarily based on motor conduction velocity tests but is now known to involve axonal and demyelination features. DI-CMTC is linked to tyrosyl-tRNA synthetase (YARS)-associated neuropathies, which are caused by E196K and G41R missense mutations and a single de novo deletion (153-156delVKQV). It is well-established that these YARS mutations induce neuronal dysfunction, morphological symptoms involving axonal degeneration, and impaired motor performance. The present study is the first to describe a novel mouse model of YARS-mutation-induced neuropathy involving a neuron-specific promoter with a deleted mitochondrial targeting sequence that inhibits the expression of YARS protein in the mitochondria. An adenovirus vector system and in vivo techniques were utilized to express YARS fusion proteins with a Flag-tag in the spinal cord, peripheral axons, and dorsal root ganglia. Following transfection of YARS-expressing viruses, the distributions of wild-type (WT) YARS and E196K mutant proteins were compared in all expressed regions; G41R was not expressed. The proportion of Flag/green fluorescent protein (GFP) double-positive signaling in the E196K mutant-type mice did not significantly differ from that of WT mice in dorsal root ganglion neurons. All adenovirus genes, and even the empty vector without the YARS gene, exhibited GFP-positive signaling in the ventral horn of the spinal cord because GFP in an adenovirus vector is driven by a cytomegalovirus promoter. The present study demonstrated that anatomical differences in tissue can lead to dissimilar expressions of YARS genes. Thus, use of this novel animal model will provide data regarding distributional defects between mutant and WT genes in neurons, the DI-CMTC phenotype, and potential treatment approaches for this disease.

Key words: nerve regeneration, tyrosyl-tRNA synthetase, YARS-associated neuropathy, YARS mutation, Charcot-Marie-Tooth Disease, adenoviral vector-mediated mouse models, neural regeneration