Quantitative and qualitative analysis of Wallerian degeneration using restricted axonal labelling in YFP-H mice
Quantitative and qualitative analysis of Wallerian degeneration using restricted axonal labelling in YFP-H mice
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DOI：
10.1016/j.jneumeth.2003.10.016
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发表时间：
2004-03-15
影响因子：
3
通讯作者：
Coleman, MP
中科院分区：
文献类型：
--
作者：
Beirowski, B;Berek, L;Coleman, MP
我们研究了 YFP-H 转基因小鼠 [Neuron 28 (2000) 41] 的有用性，该小鼠在有限的神经元子集中表达黄色荧光蛋白 (YFP)，以研究 PNS 中的华勒变性。 YFP 阳性轴突和髓鞘碱性蛋白 (MBP) 免疫细胞化学的定量显示，YFP 随机分布于约 3% 的有髓运动和感觉纤维。轴切术诱导的华勒变性表现为单个 YFP 阳性轴突中荧光信号的碎片，其形态和时间与更传统方法观察到的华勒变性相似。在共表达高剂量 Wld 的 YFP-H 转基因小鼠中，Wld 是一种嵌合基因，可以防止华勒变性 [Nat Neurosci. 4 (2001) 1199]，坐骨神经轴突切除术后远端胫神经的轴突断裂大约延迟了 10 倍。在神经外植体培养物中也观察到使用相同转基因表达系统的沃勒变性的显着延迟，从而能够对轴突碎片进行体外实时成像。值得注意的是，利用共焦荧光成像，可以在外周神经中追踪长达 2.9 厘米的单个 YFP 标记轴突。总之，转基因 YFP-H 小鼠被证明是研究体内和体外沃勒变性机制的有价值的工具。 (C) 2003 Elsevier B.V. 保留所有权利。
We investigated the usefulness of YFP-H transgenic mice [Neuron 28 (2000) 41] which express yellow fluorescent protein (YFP) in a restricted subset of neurons to study Wallerian degeneration in the PNS. Quantification of YFP positive axons and myelin basic protein (MBP) immunocytochemistry revealed that YFP was randomly distributed to approximately 3% of myelinated motor and sensory fibres. Axotomy-inducedWallerian degeneration appeared as fragmentation of fluorescent signals in individual YFP positive axons with a morphology and timing similar to Wallerian degeneration observed by more traditional methods. In YFP-H transgenic mice co-expressing a high dosage of Wld(s), a chimeric gene that protects from Wallerian degeneration [Nat Neurosci. 4 (2001) 1199], axonal fragmentation in distal tibial nerves after sciatic nerve axotomy was approximately 10 times delayed. Considerable retardations of Wallerian degeneration using the same transgenic expression system were also observed in cultures of nerve explants, enabling in vitro real-time imaging of axonal fragmentation. Remarkably, single YFP-labelled axons could be traced in peripheral nerves for unusually long distances of up to 2.9 cm exploiting confocal fluorescence imaging. Altogether transgenic YFP-H mice prove to be a valuable tool to study mechanisms of Wallerian degeneration in vivo and in vitro. (C) 2003 Elsevier B.V. All rights reserved.