Key Laboratory of Tumor Microenviroment and Neurovascular Regulation, Nankai University School of Medicine, Tianjin 300071, China.
State Key Laboratory of Medicinal Chemical Biology, College of Life Science, Nankai University, Tianjin 300071, China.
Biol Open. 2014 Dec 19;4(1):62-8. doi: 10.1242/bio.201410736.
Demyelinating diseases consist of a variety of autoimmune conditions in which the myelin sheath is damaged due to genetic and/or environmental factors. During clinical treatment, some patients undergo partial remyelination, especially during the early disease stages. However, the mechanisms that regulate demyelination remain unclear. The myelin structure, myelin formation and myelin-related gene expression are highly conserved between mammals and zebrafish. Therefore, the zebrafish is an ideal model organism to study myelination. In this study, we generated a transgenic zebrafish Tg(mbp:nfsB-egfp) expressing a fusion protein composed of enhanced green fluorescent protein (EGFP) and NTR from the myelin basic protein (mbp) promoter. Tg(mbp:nfsB-egfp) expressed NTR-EGFP reproducibly and hereditarily in oligodendrocytes along the spinal cord. Treatment of zebrafish larvae Tg(mbp:nfsB-egfp) with metronidazole (Mtz) resulted in the selective ablation of oligodendrocytes and led to demyelination, accompanied by behavioral changes, including decreased total movement distance, velocity, total movement time and fast movement time. After withdrawal of Mtz for a seven day recovery period, the expression of EGFP and MBP protein was observed again which indicates remyelination. Additionally, locomotor capacity was restored. Collectively, Tg(mbp:nfsB-egfp), a heritable and stable transgenic line, provides a novel, powerful tool to study the mechanisms of demyelination and remyelination.
脱髓鞘疾病包括多种自身免疫性疾病，其中髓鞘由于遗传和/或环境因素而受损。在临床治疗中，一些患者会发生部分髓鞘再生，尤其是在疾病早期阶段。然而，调节脱髓鞘的机制仍不清楚。哺乳动物和斑马鱼之间的髓鞘结构、髓鞘形成和髓鞘相关基因表达高度保守。因此，斑马鱼是研究髓鞘形成的理想模型生物。在这项研究中，我们生成了一种表达由髓鞘碱性蛋白（mbp）启动子驱动的增强型绿色荧光蛋白（EGFP）和 NTR 的融合蛋白的转基因斑马鱼 Tg(mbp:nfsB-egfp)。Tg(mbp:nfsB-egfp) 在沿脊柱的少突胶质细胞中可重复且遗传表达 NTR-EGFP。用甲硝唑（Mtz）处理斑马鱼幼虫 Tg(mbp:nfsB-egfp)可选择性地消融少突胶质细胞，导致脱髓鞘，并伴有行为变化，包括总运动距离、速度、总运动时间和快速运动时间减少。在停止 Mtz 治疗并进行为期七天的恢复期后，再次观察到 EGFP 和 MBP 蛋白的表达，这表明发生了髓鞘再生。此外，运动能力得到恢复。总之，Tg(mbp:nfsB-egfp) 是一种可遗传且稳定的转基因系，为研究脱髓鞘和髓鞘再生的机制提供了一种新的、强大的工具。
