Use of self-complementary adeno-associated virus serotype 2 as a tracer for labeling axons: implications for axon regeneration.
Use of self-complementary adeno-associated virus serotype 2 as a tracer for labeling axons: implications for axon regeneration.
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DOI：
10.1371/journal.pone.0087447
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Smith GM
中科院分区：
文献类型：
--
作者：
Liu Y;Keefe K;Tang X;Lin S;Smith GM
有多种示踪剂可用于轴突再生，但它们需要额外的操作示踪剂注射，耗时的免疫组织化学分析，并导致非特异性标记。在过去的几年里，大量的努力探索了其他方法，特别是使用病毒载体来研究损伤后的轴突再生。近年来的研究表明，自补充性腺相关病毒(ScAAV)具有较高的转导效率和较快的转基因表达速度。在此，我们首次使用scAAV2-GFP标记正常和损伤动物模型中皮质脊髓束(CST)、红脊束(RST)和背根节(DRG)中央轴突的长投射轴突。我们发现scAAV2-GFP可以有效地转导感觉运动皮质、红核和背根节中的神经元。强的GFP表达可沿轴突顺行运输，分别标记CST、RST和DRG中央轴突的大量轴突纤维。在共标记切片上比较scAAV2载体和单链(Ss)AAV2载体，表明scAAV2载体在DRG神经元及其轴突中诱导的转基因表达比ssAAV2载体更快和更强。在脊髓损伤和背根挤压伤模型中，scAAV-GFP可以有效地标记损伤腔周围和背根进入区(DREZ)周围的损伤轴突和再生轴突。此外，scAAV2-GFP载体可以与传统示踪剂结合，特异性地标记脊髓损伤后的感觉轴突和运动轴突。因此，我们认为使用scAAV2-GFP作为示踪剂是研究损伤后轴突再生的一种更有效和有效的方法。
Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of viral vectors, to investigate axon regeneration after injury. Recent studies have demonstrated that self-complementary Adeno-Associated Virus (scAAV) induced a high transduction efficiency and faster expression of transgenes. Here, we describe for the first time the use of scAAV2-GFP to label long-projection axons in the corticospinal tract (CST), rubrospinal tract (RST) and the central axons of dorsal root ganglion (DRG) in the normal and lesioned animal models. We found that scAAV2-GFP could efficiently transduce neurons in the sensorimotor cortex, red nucleus and DRG. Strong GFP expression could be transported anterogradely along the axon to label the numerous axon fibers from CST, RST and central axons of DRG separately. Comparison of the scAAV2 vector with single-stranded (ss) AAV2 vector in co-labeled sections showed that the scAAV2 vector induced a faster and stronger transgene expression than the ssAAV2 vector in DRG neurons and their axons. In both spinal cord lesion and dorsal root crush injury models, scAAV-GFP could efficiently label the lesioned and regenerated axons around the lesion cavity and the dorsal root entry zone (DREZ) respectively. Further, scAAV2-GFP vector could be combined with traditional tracer to specifically label sensory and motor axons after spinal cord lesion. Thus, we show that using scAAV2-GFP as a tracer is a more effective and efficient way to study axon regeneration following injury.
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影响因子：
5.3
作者：
Hu, Xinhua;Cai, Jie;Yang, Jun;Smith, George M.
通讯作者：
Smith, George M.
DOI：
10.1038/mt.2008.166
发表时间：
2008-10
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
--
作者：
通讯作者：
--
影响因子：
5.1
作者：
McCarty, DM;Monahan, PE;Samulski, RJ
通讯作者：
Samulski, RJ
影响因子：
5.4
作者：
Ferrari, FK;Samulski, T;Samulski, RJ
通讯作者：
Samulski, RJ
影响因子：
48
作者：
Marks, William J., Jr.;Bartus, Raymond T.;Olanow, C. Warren
通讯作者：
Olanow, C. Warren